Autologous Bone Marrow Transplantation - Blog Science Connections

Occult Breast Cancer Cells in Marrow 499
Cytologic and Immunocytochemical Techniques
Cells obtained directly from the harvest screens and cells harvested from
the long-term cultures were spun onto precleaned glass slides using a
Shandon Cytospin centrifuge (Shandon Southern Instruments, Inc.,
Sewickley, PA). The slides were air dried and stained with Wright's stain.
Additional slides were fixed for 2 minutes in ice-cold acetone and stained with
an avidin-biotin immunoperoxidase technique using antibodies to alphalactalbumin
or epithelial membrane antigen (EMA), antibodies # A579 and
*M613 (Dako Corporation, Santa Barbara, CA), respectively.
RESULTS
The results of applying culture techniques to detect occult tumor cells in
marrow harvests from nine consecutive patients about to undergo high-dose
therapy plus ABMT for breast cancer are summarized in Table 1. All of these
samples were obtained from patients whose marrow was considered histologically
free of malignant cells. Four of the nine samples showed no
evidence of occult tumor cells by any of the methods employed. However,
four other samples showed abnormal-appearing cells or colonies in Wright's
stained plates of the human pluripotent hematopoietic colonly-forming units
(CFCJ-GEMM) assay. Of particular note were isolated cells or large colonies
containing cells with a round or oval acidophilic, euchromatic nucleus and
occasional prominent nucleoli. All cultures yielding such cells were sampled
Table 1. Detection of Occult Tumor Cells in Cultured Marrow Harvests From
Breast Cancer Patients
Harvest
ID#
Occult Tumor Cells
Suspected/Method
Total
Weeks in
Culture
Before
Detection
Cytospin Confirmation
H & E
Stain
4 No/— No No
14 Yes/GEMM assay 6 Yes Yes
19 Yes/GEMM assay 14 Yes Yes
25 Yes/GEMM assay 6 No No
49 No/— — No No
50 Yes/GEMM assay 2 Yes Yes
54 No/— No No
141 No/— No No
EMA/cr-Lactalbumin
Immunocytochemistry
142 Yes/Culture 4 No ? Yes (see text)
Abbreviations: H & E, hematoxylin and eosin; EMA, epithelial membrane antigen;
GEMM, granulocyte, erythrocyte, monocyte, and megakaryocyte colony-forming unit.