Systems

Serumfree 

Systems 

Serumfree 

Systems 

Immunology 

Media Sera 

Reagents Molecular 

Biology 

Cytokines/ 

Growth 

Factors 

Disposables 

for Science/ 

Services 

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Serumfree Cell Culture Systems 

Cell culture, the cultivation of cells isolated from live tissue in vitro (in the test tube), is an acknowledged 

and valuable tool in the biomedical research for the determination of reproducible 

data. Apart from that, by means of cell cultures more and more highly effective substances are 

produced large-scale for medicine and research (insulin, growth factors, monoclonal antibodies). 

The Cell Culture with Serum 

Cells in vitro need nutrient solutions, so-called media which guarantee an imitation as exact as 

possible of the situation in vivo (in live organism). On the other hand these media – mixtures of 

nutrients, salts, trace elements, buffer substances, growth factors, protective substances and 

many other components – must be complemented for this purpose by mostly natural, highly complex 

additives. Not long ago animal but also human serum were the means to be chosen for reasons 

of production technology and also for lack of alternatives. 

Function of Serum in the Cell Culture 

• Hormone factors stimulate cell growth, proliferation and differentiation 

• Attachment factors favour or enable the attachment of the cells to the culture dish (Biomatrix) 

• Transport and binding proteins take care among other things of the supply of hormones, 

minerals and lipids 

• Serum proteins bind toxic substances 

This serum, mostly fetal bovine serum (FCS), is problematic for several reasons. 

Disadvantages of Serum in the Cell Culture 

• The composition of serum is not constant and varies with the age of the foetus, with the 

origin and feeding of the animals and with the time of the year. 

• Serum batches have to be tested for their suitability before use. 

• Test results are often not convincing and often not comparable because of the undefined 

and inconstant composition of the serum. 

• Risk of a contamination with bacteria, fungi, mycoplasm and virus from serum 

• Risk of a contamination with TSE-agents (transmissable spongioform encephalopathy) 

• Possibility of a contamination of the end product with serum proteins or pyrogens 

• Time-consuming cleaning of the end products from culture media containing serum 

• Availability and costs of the serum 

The Serumfree Cell Culture 

Because of the numerous disadvantages of the cell culture with nutrient media containing serum, 

tests have been made for quite some time to establish cell cultures under serumfree conditions. 

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Advantages of a Serumfree Cell Culture 

• Lower risk with regard to a contamination with bacteria, fungi or virus 

• Better defined and reproducible formulations allow more convincing and comparable 

research results 

• Time-consuming batch tests are not necessary 

• Elimination of a source for possible infectious agents (virus, prions) 

• Facilitation of the cleaning of the end products 

• Fulfilment of legal conditions for the production of medical products 

• Reduction of contaminations of the end products by culture residues 

Serumfree Culture Media and Serum Substitutes 

from PAN-Biotech 

In series of tests for many years PAN-Biotech has developed, optimized and successfully 

marketed a number of serumfree media for the most different cell types. 

The development and optimization of serumfree media takes a lot of time and expenditure. 

Several months can often pass until the release of a new product. 

With the development of a fully-automatic cell culture system (patent pending) PAN-Biotech 

pursues a completely different path. With this system they have succeeded in cultivating different 

samples simultaneously under identical conditions and under constant microscopic observation. 

By adding individual components res. by changing the concentration, a positive (improvement of 

growth) res. negative effect (inhibition of growth) can directly be measured. All relevant data can 

be stored and recalled later. Morphologic changes are identified immediately and evaluated 

depending on the media composition. 

The cell culture system offers many advantages for the development of serumfree media: 

• Reduction of the development periods by more convincing results 

• Adjusting the optimal concentration of individual substances 

• Comparison measurements of competitor products and recording the results 

• Culture conditions (temperature, CO2-fumigation) can be changed infinitely variably at any time 

• Complete control of individual batches 

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Fully-Automatic Cell Culture System from PAN 

Incubation chambers with connections 

for media and gas supply 

CO2-supply 

Heating plate 

Glass top for microscopic observation 

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Media supply 

Digital camera 

Incubation chambers 

Control heating 

CO2-fumigation 

Image processing 

In the following example, CHO-cells were cultivated in DMEM with 4,5 g/l glucose with 

L-glutamine, with pyruvate and different serum substitutes (seed 1 x 10 4 /ml, evaluation 7th day, 

cells not adapted). 

CHO with serum substitute 1 CHO with serum substitute 2 CHO with 5 % Panexin 

Cells do not form any Cells do not form any Cells form uniform 

cell layer, loose attachment cell layer, loose attachment cell layer 

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In order to satisfy our customers’ wishes, we will continue to expand and optimize our range of 

serum- and proteinfree media. 

Beside nutrient solutions ready for use (PANSERIN TM ) we can also offer serum substitutes (PANEXIN) 

to our customers which are added in a concentration of 2 % - 8 % instead of the serum to the 

respective basic media (e.g. RPMI 1640, DMEM, DMEM/F12). 

Serumfree nutrient media without animal or human components res. proteinfree media are also 

already available at PAN-Biotech res. in development. 

In many cases an adaption of the cells to the serumfree culture is not necessary; with 

critical cells (primary cells) the cells are slowly familiarized with the serumfree conditions 

with help of our detailed records. 

Product Survey 

Product: suitable for: Quantity: Cat.-No.: Page: 

PANSERIN TM 401 serumfree allround medium 500 ml P04-710401 7-9 

Serumfree allround for many cell types 100 ml P04-710401M 

medium 


Serumfree allround for cells which also need Insulin 100 ml P04-710411M 

medium with Insulin 


Serumfree allround for cells that hardly attach 100 ml P04-710412M 

medium with Fetuin 

PANSERIN TM 413 serumfree cultivation 500 ml P04-710413 10-11 

Growth factors of T-cells 

mixture separate 

PANSERIN TM 415 serumfree cultivation 500 ml P04-710415 12 

of Osteoblasts 100 ml P04-710415M 

PANSERIN TM 416 Kit serumfree cultivation 500 ml P04-710416 13-15 

Basic medium with of DC (Dendritic Cells) 100 ml P04-710416M 

growth factor set 

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Product Survey 

Product: suitable for: Quantity: Cat.-No.: Page: 

PANSERIN TM 604ST serumfree medium for CHO- 500 ml P04-604ST 16-18 

cells in suspension culture 

PANSERIN TM 604S serumfree medium for CHO- 500 ml P04-604S 16-18 

cells in suspension culture. 

Medium doesn’t contain any 

components of human or 

animal origin 

PANSERIN TM 604SPF proteinfree medium for CHO- 500 ml P04-604SPF 16-18 

cells in suspension culture – 

Medium with chemically 

defined substances 

PANSERIN TM 604 serumfree medium for CHO- 500 ml P04-710604 16-18 

cells in adherent culture 100 ml P04-710604M 

PANSERIN TM PX10 serumfree medium for 500 ml P04-710PX10 19-21 

myeloma cells, production 100 ml P04-710PX10M 

of monoclonal Antibodies 

PANEXIN many adherent and non- 100 ml P04-95100 22-23 

adherent cells – serum substitute 10 ml P04-95010 

PANEXIN H many adherent and non- 100 ml P04-95300 22-23 

adherent cells – serum substitute 10 ml P04-95030 

without animal components 

PANEXIN D adherent cells that hardly 100 ml P04-95200 22-23 

attach – serum substitute with 10 ml P04-95020 

additional attachment factors 

PANEXIN S many adherent and non- 100 ml P04-95400 22-23 

(in development) adherent cells – serum substitute 10 ml P04-95040 

without animal or human 

components 

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PANSERIN TM 401 

PANSERIN TM 401 is a complete medium ready for use for the serumfree cultivation of a multitude 

of cells. Among others the following cells have been cultivated successfully: 

• Hybridoma 

• Lymphocytes 

• Macrophages 

• Fibroblasts 

• Melanocytes 

• Carcinome cells 

• HEK-cells 

• HeLa-Zellen 

• CHO-cells 

Composition: 

Based on Iscove’s MEM, trace elements, albumin, cholesterol, soya lipids and vitamins were added 

to the medium. It does not contain any growth or attachment factors or any insulin. 

Application: 

For many cells (SP2, HEK, L929, CHO) a slow adaption to PANSERIN TM 401 is not necessary. 

They are adapted to the serumfree culture according to the 

Direct Adaption Method. 

• Transfer sound cells (> 90 % Vitality) of the logarithimic growth phase from the culture containing 

serum directly into PANSERIN TM 401. A higher cell seed (5 x 10 4 - 1 x 10 5 ) facilitates the adaption 

to the serumfree culture. 

• After 36 - 48 hours in the culture, replace medium by fresh PANSERIN TM 401. 

• As soon as the cells have confluently grown at 90 % to 100 %, passage the cells with the usual 

trypsinating technique. You have to be aware that cells in the serumfree culture show an 

extremeley sensitive reaction on trypsin. Normal trypsin concentrations (0,25 %) can be used, 

however the incubation period should be as short as possible at 4 °C. The trypsin can be 

removed by washing the cells and centrifugation. Soybean-trypsin-inhibitor has to be used with 

caution as it is toxic for some cells. 

• After several passages into PANSERIN TM 401 the adaption is completed. 

Critical cells can be adapted to the serumfree culture according to the 

Gradual Adaption Method. 

• Transfer sound cells (> 90 % Vitality) of the logarithmic growth phase from the culture containing 

serum (10 % - 20 % FCS) directly into PANSERIN TM 401. Reduce serum contents to 5 %. The 

usual seeding densities can be kept, but with very critical cells increase seeding densities 

by 1,5 to 2 x. 

• After 2 passages with 5 % FCS supplementation, reduce FCS contents to 1 %. 

• After 2 - 3 passages with 1% FCS, reduce serum contents to 0,5 %. 

• After further 2 - 3 passages with 0,5 % FCS, cultivate cells without addition of FCS 

in PANSERIN TM 401. 

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PANSERIN TM 401 does not contain any attachment factors. With some cell types a pretreatment 

of the incubation dishes with gelatin, collagen, poly-D-lysine or fibronectin can considerably facilitate 

the culture under serumfree conditions or even enable it. 

Please note this above all for low seeding densities. 

Additional growth factors are necessary for some cells and have to be added depending on the 

demand. Please note another important point: 

With every adaption to serumfree media, changes of the cells should be taken into consideration. 

These changes can concern the morphology, the karyotype, the surface marker etc. Thus cells in 

serumfree medium don’t always have to be identical with those from the culture containing serum 

in which they originate (selection). 

SP2/0-Ag-14 in PANSERIN TM 401 

Cells were cultivated in 

PANSERIN TM 401 without adaption 

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L 929 in PANSERIN TM 401 

Cells were cultivated in 

PANSERIN TM 401 without adaption phase. 

8 

Typical growth curve L929 

in PANSERIN TM 401: 


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L929 were cultivated in 


without adaption phase 

As a comparison L929 in DMEM 

with 4,5 g/l glucose with 10 % FBS

Order Information: 

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Typical growth curve 

SP2/0-Ag-14 

in PANSERIN TM 401: 

SP2/0-Ag-14 cells were 

cultivated in PANSERIN TM 401 

without adaption phase 

As a comparison cell growth 

in RPMI 1640 with 10 % FBS 

Product: suitable for: Quantity: Cat.-No.: 

PANSERIN TM 401 serumfree allround medium 500 ml P04-710401 

Serumfree allround for many cell types 100 ml P04-710401M 

medium 


Serumfree allround for cells which also need Insulin 100 ml P04-710411M 

medium with Insulin 


Serumfree allround for cells that hardly attach 100 ml P04-710412M 

medium with Fetuin 

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PANSERIN TM 413 is a medium ready for use for the cultivation of lymphocytes (from full blood). 

There is a multitude of questions with the subject matter of short-term cultivation of human cells. 

As human full blood is relatively easily accessible and because of the possibility of preparing 

peripheral lymphocyte cultures for a short time, quite a lot of methods have been developed in 

order to keep blood cells and especially lymphocytes in culture. In principle blood cells in the culture 

die relatively soon, only lymphocytes can be kept in culture over several cell divisions. However, 

in order to reach a division of the cells which are normally not proliferating, the cells must be stimulated 

with specific mitogens. These mitogens are mostly plant lectines (phytohaemagglutinin, PHA). 

Composition: 

Based on Iscove’s MEM, trace elements, albumin, cholesterol, soya-lipids and vitamins were 

added to the medium. A growth factor mixture is also supplied which has to be added to the medium 

shortly before the cultivation. 

Application: 

1. Isolation of lymphocytes from full blood by means of density gradient centrifugation 

• Mix heparinised blood 1: 1 with PBS and add it into a centrifuge tube which has been filled with 

lymphocyte separating medium (Pancol density 1,077 g/ml) before: pipette carefully in order 

to avoid a phase mixture! 

• Centrifuge the gradient at 400 x g for 30 minutes at ambient temperature (brake of the centrifuge 

set on “off”); 4 phases are created 

– top phase plasma 

– opaque whitish bands (lymphocytes) 

– separating medium 

– pellet with erythrocytes and granulocytes 

• Suck off the plasma with a pipette and transfer the lymphocytes with another pipette into a new 

centrifuge glass. 

• Wash the lymphocytes with PBS (without Ca, Mg) and centrifuge them at 100 x g for 10 minutes. 

Repeat washing step. 

2. Cultivation and stimulation of the lymphocytes 

• Resuspend lymphocytes in PANSERIN TM 413. 

• For the stimulation of the lymphocyte proliferation adjust the cells to a cell density of approx. 

1x10 5 and add phytohaemagglutinin in a concentration of 1-5mg/ml; the incubation time is 

48 to 72 hours, depending on the kind and origin of the lymphocytes and depending on the 

further use. 

• Further cultivation in PANSERIN TM 413. After approx. 14 days a restimulation is necessary. 

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PANSERIN TM 413 serumfree cultivation 500 ml P04-710413 

Growth factors of T-cells 

mixture separate 

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PANSERIN TM 415 is a complete medium ready for use for the cultivation of osteoblasts. 

These osteoblasts are of mesenchymal origin as they are bone-building cells. 

They lie almost always next to each other epithelium-like in a layer and have contact to each 

other via short delicate appendices. 

The osteoblasts produce the intercellular substance 

of the bone tissue, which consists of basic substance 

and collagen fibres (type I). 

Because of the matrix production continuing during 

the bone development and the bone growth, the 

osteoblasts, which have first been close neighbours, 

drift apart more and more. Thus the conditions for the 

metabolism and the matrix production get increasingly 

worse which is finally stopped completely. 

The conversion into osteocytes takes place. 

Composition: 

PANSERIN TM 415 consists of nutrient-rich basal medium to which lipids, albumin, transferrin, insulin 

and trace elements have been added for an optimal growth of osteoblasts. 


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Osteoblasts in PANSERIN TM 415 

Osteoblasts were cultivated without 

adaption from a culture containing 

serum directly in PANSERIN TM 415. 

As a comparison the growth in DMEM 

with 4,5 g/l glucose with L-glutamine, 

pyruvate and 10 % FBS 


PANSERIN TM 415 serumfree cultivation 500 ml P04-710415 

of Osteoblasts 100 ml P04-710415M 

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PANSERIN TM 416 is a serumfree medium (basic medium) which is, after supplementation of growth 

factors, suitable for the production of dendritic cells. 

Dendritic cells are highly specialized antigen-presenting cells and can initiate and regulate antigenspecific 

immunoresponses. This ability can be used in order to generate immunoresponses against 

certain proteins of tumour cells and thus to fight against tumours with the immune system. Dendritic 

cells have been isolated from a great variety of non-lymphatic and lymphatic tissue of human 

beings, mice and other species. 

For the generation of tumour vaccines, dendritic cells can be produced from the peripheral blood 

of tumour patients. In clinical studies the principal effectiveness of a vaccination with dendritic 

cells has been shown. 

Antigen Uptake: 

In almost any tissue of the body, dendritic cells form a dense network of guardian cells that take 

up extracellular components by processes such as phagocytosis and endocytosis and thus analyse 

their environment. Proteins that have been taken up undergo an intracellular decomposition into 

peptides, are bound to MHC-molecules and transported to the cell surface. Thus antigenic 

determinants of the peptides are made recognizable to T-cells. Within the scope of the physiological 

cell regeneration dendritic cells leave the peripheral tissue and migrate with the lymph into a 

regional lymph node where they interact with T-cells. From an intact tissue dendritic cells reach 

the lymph node in unactivated condition. 

A functioning monitoring system excels in the ability to quickly and specifically recognize damaging 

processes and to take suitable steps against them. For this purpose, dendritic cells carry receptors 

on their surface for a variety of danger signals which can be radiated from microorganisms, 

mediators inherent in the body or activated T-cells. Examples for microbial structures activating 

dendritic cells are lipopolysaccharides of Gram-negative bacteria, cytidine guanosine dinucleotide 

(cpG) – rich bacterial DNA and viral doublestranded RNA. Endogenous mediators, for which dendritic 

cells have special receptors and which send an activating signal, are cytokines, prostanoids and 

adenine nucleotides. Activated T-cells can stimulate dendritic cells by means of the CD40-ligand 

integrated in their cell membrane. The activation of these different receptors induces essential 

cell changes which are summarized by the term maturation. The ability of phagocytosis gets lost. 

Peptides bound to MHC-molecules are presented in a higher density and with greater stability. The 

cytoskeleton structure is reorganized and a changed expression of chemokine receptors enables 

the dendritic cells to get from the focus of inflammation into the draining lymph node. Co-stimulating 

molecules on the surface of dendritic cells and the release of cytokines, e.g. interleukin-12, finally 

allow the dendritic cells an efficient interaction with T-cells. 

Induction of an Immunoresponse: 

In the lymph node, dendritic cells interact with various lymphocyte populations. Above all T-cells, 

which haven’t yed had any antigen contact, feel the cell surface of dendritic cells and are activated 

if the T-cell receptor recognizes the presented antigen. This central process for the acquired (antigenspecific) 

immunoresponse is called “Priming”. Cytotoxic T-cells develop from CD8 cells and these 

cytotoxic T-cells are able to kill those cells which they recognize with their T-cell receptor. 

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Tumour Vaccination with Dendritic Cells: 

Tumour cells express specific proteins which can be recognized as antigenic determinants by 

T-cells. As a rule, however, this is not sufficient for the immune system to generate an effective 

immunoresponse against tumour cells; there is rather a tolerance. On the one hand this is because 

tumour-associated antigens are often also found in sound tissue in a low density; on the other 

hand tumour cells have numerous strategies to escape an immunoresponse. In a number of animal 

experiments, however, it could be clearly shown that this tolerance for tumours can be broken by 

a vaccination with dendritic cells. 

Generation of Dendritic Cells: 

Dendritic cells are derived from hematopoetic precursor cells in the bone marrow. Three different 

subpopulations with each characteristic features and functions are described for the human being: 

myeloid dendritic cells, plasmacytoid and Langerhans cells of the skin. For tumour vaccinations, 

myeloid dendritic cells are mainly of interest as they are especially capable of taking up and 

presenting antigens. Dendritic cells with myeloid characteristics can be produced by an in vitro 

culture of monocytes in the presence of the cytokines interleukin-4 (IL-4) and granulocytesmacrophages 

colony-stimulating factor (GM-CSF). Alternatively dendritic cells can be generated 

from CD34+ hematopoetic stem cells of the peripheral blood. 

By the addition of growth factors, e.g. Klt3-ligand, dendritic cells in the blood, which normally make 

up only approx. 0,1 - 0,5 % of the mononuclear cells, can be expanded many times over. 

After activation, dendritic cells reach their full capacity for the T-cell stimulation. For the maturation 

of the dendritic cells, cytokines (TNF alpha), LPS or monocyte-conditioned medium are used. 

Production and serumfree cultivation of dendritic cells from mononuclear cells 

of peripheral blood (PBMC) 

PAN-Biotech has developed an optimised medium for the serumfree cultivation of dendritic cells. 

PANSERIN TM 416 (basic medium) is supplemented by the growth factors delivered with it and thus 

allows the serumfree cultivation of dendritic cells. 

Separation of Blood with Separating Medium (Pancoll 1,077 g/ml) 

The blood samples should be processed as soon as possible after production in order to 

achieve optimal results. A storage of the blood samples for 24 hours at ambient 

temperature causes among other things a reduced output of lymphocytes, a change of 

the surface markers and a reduced response on mitogen stimulation. 

1) Add Pancoll (3 ml) into a suitable, sterile centrifuge tube under sterile conditions. 

2) Carefully coat the separating medium with the diluted blood sample (4 ml). Important: Don’t mix the blood sample with Pancoll! 

3) Centrifugation at 400 x g for 30 - 40 minutes at 18 - 20 °C. 

4) After the centrifugation, carefully take off the upper phase (containing serum and platelets) with a pipette without mixing the 

interphase with the lymphocytes. 

5) Transfer the lymphocyte band into a new centrifuge tube with a new pipette. Here it is important to take off the whole material 

of the interphase with as little volume as possible. 

6) Add at least 3 volumes of a physiological saline solution (6 ml) to the lymphocytes. 

7) Carefully suspend the lymphocytes with a pipette. 

8) Centrifugation at 60 - 100 x g for 10 minutes at 18 -20 °C. 

9) Reject the supernatant. 

10) Repeat washing step (point 6 - 9). 

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Serumfree Cultivation of Dentritic Cells in PANSERIN TM 416 

After the last washing step the mononuclear cells are transferred with a cell density of 1 x 10 7 

cells/ml into PANSERIN TM 416. In order to remove non-adherent cells, the culture dishes are put 

into the incubator for 2 hours. Then the supernatant is carefully taken off and replaced by new 

PANSERIN TM 416. GM-CSF (10 ng/ml) and interleukin-4 (10 ng/ml) are added as growth factors. 

The culture dishes are incubated for further 6 days in the incubator and every day half of the 

medium is replaced by new medium which is supplemented by GM-CSF and IL-4. 

Maturation of Dentritic Cells 

The dendritic cells are suspended with a density of 10 6 cells/ml in PANSERIN TM 416 with GM-CSF 

and IL-4. For the maturation of the cells, 50 ng/ml LPS or TNFalpha (20 ng/ml) are added. 

The cells can be harvested after approx. 3 days. 

Dendritic cells in PANSERIN TM 416 

with GM-CSF and IL-4 (6th day) 


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Dendritic cells in RPMI 

with 10 % FCS, GM-CSF and IL-4 (6th day) 


PANSERIN TM 416 Kit serumfree cultivation 500 ml P04-710416 

Basic medium with of DC (Dendritic Cells) 100 ml P04-710416M 

growth factor set 

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PANSERIN TM 604 is a complete medium ready for use for the cultivation of transfected and 

non-transfacted CHO-cells (Chinese Hamster Ovary). 

These cells are frequently used for the production of recombinant proteins. The cells normally grow 

adherently (e.g. in roller cultures), however with suitable media and with the optimal media they 

can also be kept in suspension culture (continuous perfusion or batch systems). 

PAN-Biotech has developed an optimal version for every application: 

For the suspension culture: 

• PANSERIN TM 604ST serumfree – optimal cell growth, quick adaption 

• PANSERIN TM 604S without protein or peptide components of animal or human origin 

• PANSERIN TM 604SPF chemically defined, proteinfree 

The following graph shows the growth curves of the PANSERIN TM 604 versions. 

The CHO-cells were cultivated from an adherent culture containing serum in PANSERIN TM 604 

without a preceding adaption. The seed amounted to 50 000 cells/ml. The maximum generation 

times were 15,3 to 18,5 hours. 

Composition: 

PANSERIN TM 604 serumfree: 

A basal medium optimized for suspension cultures, which had been designed for a high cell density, 

was supplemented with hormons, lipids and special growth factors for an optimal growth of CHOcells. 

It does not contain any undefined lysates or plant hydrolysates. It allows a very quick growth 

of the cells without long adaption phases. 

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PANSERIN TM 604S without animal or human components 

Basal medium which has been completed with exclusively recombinant proteins. It does not contain 

any undefined lysates or plant hydrolysates. Especially suitably for the production of proteins which 

are applied in sensitive fields. It excels by a quick cell growth without long adaption phases. 

PANSERIN TM 604SPF proteinfree, chemically defined: 

This medium is completely proteinfree and chemically defined. After a short adaption phase excellent 

growth rates can be reached with this medium and these growth rates are only slightly under the 

culture media containing protein and serum. Especially suitable for the production of recombinant 

proteins for the application in sensitive fields. 

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CHO-Zellen in PANSERIN TM 604ST, serumfree 

CHO-Cells were cultivated directly without adaption 

phase from an adherent culture containing serum 

in a suspension culture with PANSERIN TM 604 

(shaking culture). 

Because of an optimal and well-balanced composition of the medium an aggregate formation of 

the cells, as it can be seen with many serumfree media, could be largely avoided (see illustration). 

For the adherent culture: 

PANSERIN TM 604 supports the growth of adherent CHO-cells in an optimal way. With attachment 

factors the cells can grow on quickly and this allows a high cell output within a short time. 

Composition: 

CHO-Cells in PANSERIN TM 604 

CHO-Cells were transferred from 

a culture containing serum directly 

into PANSERIN TM 604. 

A basal medium optimized for CHO-cells was supplemented with albumin, insulin, transferrin, 

attachment factors, growth factors, lipoproteins and trace elements. It does not contain any 

undefined hydrolysates or lysates. 

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CHO-Cells were cultivated from a culture containing serum in PANSERIN TM 604 without adaption. 

As a comparison CHO-cells in DMEM with 4,5 g/l glucose, L-glutamine and pyruvate, supplemented 

with 10 % FBS. 



For the suspension culture 

PANSERINTM 604ST serumfree medium for CHOcells 

in suspension culture 

500 ml P04-604ST 

PANSERIN TM 604S serumfree medium for CHO- 500 ml P04-604S 

cells in suspension culture. 

Medium doesn’t contain any 

components of human or 

animal origin 

PANSERIN TM 604SPF proteinfree medium for CHO- 500 ml P04-604SPF 

cells in suspension culture – 

Medium with chemically 

defined substances 

For the adherent culture 

PANSERIN TM 604 serumfree medium for CHO- 500 ml P04-710604 

cells in adherent culture 100 ml P04-710604M 

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PANSERIN TM PX10 

PANSERIN TM PX10 is a serumfree medium ready for use for the production of monoclonal 

antibodies. Due to the optimised composition the cells are stimulated to proliferationeven with 

low seed densities. 

SP2/0-Ag-14 in PANSERIN TM PX10 SP2/0-Ag-14 in RPMI 1640 with 10 % FCS 

SP2/0-Ag-14 have been transferred from a culture containing serum (10 % FCS in RPMI 1640) 

directly into PANSERIN TM PX10 and cultivated in the incubator at 37 °C and with 5 % CO2-fumigation 

for 5 days. 

Seed 1000 cells/ml. The cells grow very well without a long adaption phase, comparable 

to a culture containing serum (picture on the right). 

PANSERIN TM PX10 distinguishes itself, beside the excellent growth-stimulating properties, by its 

cloning capacity. Conventional serumfree systems often need lengthy and timeconsuming adaption 

steps and cell seeds up to 10 5 cells/ml. 

By contrast, most clones can be directly transferred into PANSERIN TM PX10. With PANSERIN TM PX10, 

clones can even be produced without great difficulties. 

Cloning of SP2/0-Ag-14 

in PANSERIN TM PX10 

Composition: 

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Cloning of SP2/0-Ag-14 

in RPMI 1640 with 20 % FCS 

PANSERIN TM PX10 contains cleaned proteins, lipids, salts, amino acids, trace elements and hormones 

in an optimised prescription. It doesn’t contain any undefined hydrolysates or lysates (e.g. 

peptones). L-glutamine is already included in the medium. Antibiotic is not included in the formulation. 

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19

spleen 

hybrid cells 

Immunization 

BIOTECH GmbH 

cell fusion 

Production of cell clones 

analysis of the antibody production 

cell culture 

production of 

monoclonal 

antibodies 

with 

Myeloma cells 

PANSERIN TM PX10 

 

analysis 

of the 

antibodies 

Production and cleaning of the monoclonal antibodies 

20 

Monoclonal Antibodies 

Antibodies are proteins specifically 

binding to structures res. 

substances which are called 

angines. They are part of the 

defense mechanisms of the 

immune system and are produced 

by the leucocytes (B-lymphocytes) 

as a reaction on the respective 

specific antigen. 

The human humoral immune 

system is able to produce a 

tremendous number of antibodies 

(approx. 10 8 ) with different 

antigen binding specificity. 

They serve to identify foreign body 

antigens (bacteria, virus). 

This diversity is brought about by 

a relatively small gene repertoire 

and a recombination of V-, J- and 

D-sequences takes effect on the 

variety of antibodies of the 

variable regions. 

The work of Köhler and Milstein 

set a landmark for the production 

of monoclonal antibodies. The 

conventional way of production 

used to bring about only polyclonal 

antibodies in small quantities but 

the specificity and presentable 

quantitiy for experimental work 

were made possible only by the 

hybridisation of cells (not 

indefinitely divisible, e.g. spleen 

cells) from immunized animals 

and the indefinitely divisible 

myeloma cells. 

Such hybrid cells can be multiplied 

further on in the culture and then 

they produce a clone (offspring 

of identical cells) by which monoclonal 

antibodies are produced. 

The field of application and the distribution of monoclonal antibodies reach from a therapeutical 

application (fight against cancer) to diagnostic applications (ELISA) and are still increasing. 

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Application: 

In many cases a serum-free cultivation can be carried out without time-consuming adaption steps. 

With critical clones, reduce the serum slowly (see instruction PANSERIN TM 401). 

• Warm up PANSERIN TM PX10 to 37 °C. 

• Transfer hybridoma directly into PANSERIN TM PX10. In most cases a cell number of 1000 cells/ml 

is sufficient. 

• Incubate the cells in the usual way in the CO2-incubator at 37 °C (5 % CO2-fumigation) res. multiply 

them in the bioreactor. 

• Extract monoclonal antibodies from the supernatant. 

Order Information 


PANSERIN TM PX10 serumfree medium for 500 ml P04-710PX10 

myeloma cells, production 100 ml P04-710PX10M 

of monoclonal Antibodies 

BIOTECH GmbH 

 

The company 


phone +49 (0) 85 43 - 60 16-30, fax +49 (0) 85 43 - 60 16-49 


21 

Typical growth curve 

of SP2/0-Ag-14 

in PANSERIN TM PX10 

Sp2/0-Ag-14 cells have been 

transferred from a culture 

containing serum (RPMI 1640 

with 10 % FCS) directly into 

PANSERIN TM PX10. 

Seed 1000 Zellen/ml. As a 

comparison Sp2/0-Ag-14 

in RPMI 1640 with 10 % FCS. 

SP2/0-Ag-14 cells were 

used from a culture 

containing serum directly 

in a cloning efficiency test 

with PANSERIN TM PX10 

The absolute cloning efficiency 

was 75 % (77 % with 20 % FCS) 

the relative cloning efficiency 

97 % (100 % with 20 % FCS)

PANEXIN 

PANEXIN is a chemically defined serum substitute for the cultivation of adherent and nonadherent 

cells under serumfree conditions. The sterile solution ready for use is added in an end concentration 

of 2 % to 8 % (dependant on the cells) to the culture medium. It supports the growth of many cell 

types in an optimal way. 

Composition: 

PANEXIN contains cleaned protein, lipids, salts, amino acids, trace elements, attachment factors 

and hormones in an optimized recipe. It does not contain any undefined hydrolysates or lysates 

(e.g. peptone). 

PANEXIN H does not contain any components of animal origin. A version, which contains neither 

animal nor human components, is at the development stage at the moment. 

Application: 

In many cases a serumfree cultivation can be carried out without time-consuming adaption steps 

(many permanent cell-lines e.g. SP2, HEK, CHO, L929). 

• Defrost PANEXIN slowly in the water bath. 

• Trypsinate adherent cells in the usual way (e.g. 0,25 % trypsin solution). As soon as the cells 

have rounded and separated from the surface (the process can be accelerated at 37 °C), 

inactivate with FCS trypsin. 

• Add the cells to PBS (without Mg and Ca) and centrifuge. Take off supernatent sterile and reject 

it. Resuspend cells in basic medium (RPMI 1640, DMEM or others) and determine cell number. 

• To the usual basic medium (e.g. RPMI 1640, DMEM, MEM, DMEM/F12) add 5 % PANEXIN sterile. 

• Add cell suspension with 5000 - 50000 cells/ml to the basic medium supplemented with PANEXIN. 

• Incubate the cells in the usual way in the CO2-incubator at 37 °C. 

• Non-adherent cells (e.g. SP2) can be directly transferred into the nutrient solution (e.g. RPMI 

1640, IMDM) supplemented with 5 % PANEXIN. 

Depending on the cell type, the optimal PANEXIN concentration can vary between 2 % and 8 %. This 

must be determined individually for each cell type. The tests should be started with 5 % PANEXIN 

concentration as with this concentration the best results have been reached with most cells. Higher 

concentrations (> 10 % PANEXIN) are already toxic for many cells and should not be used. 

As basic medium you can use the classical standard media like RPMI 1640, DMEM (high or low 

glucose), DMEM/F12, IMDM and so on. u.s.w. Pay attention that L-glutamine is available in a 

sufficient quantity (maybe more has to be added). Also here, depending on the cell type, differences 

with the different standard media can be seen. We have achieved the best results with RPMI 1640 

and IMDM for non-adherent cells and with DMEM and DMEM/F12 for adherent cells, in combination 

with 5% PANEXIN. 

For critical cells (e.g. primary cells) an adaption to PANEXIN is necessary. 

Adaption Instructions for PANEXIN 

Condition for a successful adaption are vital cells (exclusion trypan blue) which should be harvested 

in the logarithmic growth phase. 

BIOTECH GmbH 

 

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phone +49 (0) 85 43 - 60 16-30, fax +49 (0) 85 43 - 60 16-49 


22

• Harvest cells in the usual way. 

• Add 5 % PANEXIN to the usual basic medium (the ready solution is stable at 4°C 

for at least 4 weeks) = MedPAN 

• Add 10 % FBS to the basic medium = MedFBS 

1. 75 % MedFBS : 25 % MedPAN 

Seed 5 x 10 4 – 1x10 5 cells/ml • Observe cells under the microscope, passage at 90 % confluence cells – 2 - 3 passages 

when the cells grow well convert into 







4. 100 % MedPAN 

Seed 5 x 10 4 – 1x10 5 cells/ml • Observe cells under the microscope 

With some cells an adaption to serumfree conditions is only hard to be reached res. not possible. 

The following measures can facilitate a successful adaption: 

• Seed with a higher cell number 

• Addition of growth factors (if known which factors have a positive effect on the cells concerned) 

• Coating of the culture dishes with fibronectin, collagen, gelatin or other attachment factors 

(in case of adherent cells) 

• Change of the basic medium 

Stability: 

PANEXIN a sterile liquid (concentrate) and is supplied in deep-frozen condition. 

Repeated defrosting and deep-freezing again should be avoided. PANEXIN is stable at 4 °C for at 

least 4 weeks. The durability is at least 24 months with a storage temperature of -20°. 

Examples of cell cultures in PANEXIN: 

CHO-cells 

5 % PANEXIN/DMEM 

HEK-cells 


SP2/0-Ag-14-cells 

5 % PANEXIN/RPMI 1640 

Primary hum. fibroblasts 


BIOTECH GmbH 

 

The company 

23 

the detailed 

Order Information is on page 6 


phone +49 (0) 85 43 - 60 16-30, fax +49 (0) 85 43 - 60 16-49 


PAN-Biotech GmbH – 

Your way to success 

PAN – BIOTECH GmbH 


phone: +49 (0) 85 43 / 60 16 – 30 

fax: +49 (0) 85 43 / 60 16 – 49 

info@pan-biotech.de; www.pan-biotech.de 

BIOTECH GmbH

Systems

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