Ex vivo enrichment of malignant carcinoma cells in primary culture.
OBJECTIVE
Establishing and maintaining human tumours in primary culture can be challenging. In this application, a short-term primary culture process is desired to ensure cells maintained in culture are representative of the in vivo tumour for the purpose of chemoresponse testing. To ensure the appropriate cells are being grown, the cultures must be evaluated for malignancy. The clinical gold standard determination of malignancy is cytological evaluation by a cytopathologist.


METHODS
Fifty human tumour specimens (breast, colon, lung, ovary) were established and maintained in primary culture. Cytospins were prepared upon initiation of culture and again at completion of the culture process. Cytospins were stained (Diff-Quik, Papanicolaou) and evaluated by a cytopathologist for the percentage of malignant cells at both times.


RESULTS
An increase in the percentage of malignant cells was noted in 86% (43/50) of the cultures evaluated; 8% (4/50) of the cultures maintained the same percentage of malignant cells throughout the culture period, and 6% (3/50) displayed a decrease in malignant cells. On average, the percentage of malignant cells increased by 37% and was not associated with the length of culture (range 5-28 days).


CONCLUSIONS
The described primary culture process enriches for malignant cells, which is desirable for further evaluation such as chemoresponse testing.
Heinzman
Jamie M
JM
Precision Therapeutics Inc., 2516 Jane Street, Pittsburgh, PA 15203, USA.
Brower
Stacey L
SL
Bush
Jason E
JE
Silverman
Jan F
JF
D002277:Carcinoma; D018929:Cell Culture Techniques / Q000379:methods*; D004354:Drug Screening Assays, Antitumor; D005455:Fluorescent Antibody Technique; D006801:Humans; D014407:Tumor Cells, Cultured / Q000502:physiology*