Comparison of three culture media for the establishment of melanoma cell lines

• Published in: Cytotechnology (Dordrecht) Vol. 62; no. 5; pp. 403 - 412
• Main Authors: Pandolfino, M. C, Saïagh, S, Knol, A. C, Dréno, B
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : Springer Netherlands 01.10.2010; Springer Netherlands; Springer; Springer Nature B.V
• Subjects: Adoptive immunotherapy; Antibodies; Antigen (tumor-associated); Antigens; Biochemistry; Biological and medical sciences; Biomedicine; Biotechnology; Cell culture; Cell surface; Cell-based therapy; Chemistry; Chemistry and Materials Science; Culture media; Cytology; Fetal calf serum; Fundamental and applied biological sciences. Psychology; Good Manufacturing Practice; Immunotherapy; Laboratories; Lymph nodes; Lymphocytes; Melanoma; Original Research; Serum-free cell culture medium; Tumor cell line growth; Tumor cell lines; Tumors; United Kingdom > UK; France; United States > US; Serum-free cell culture medium; Cell-based therapy; Melanoma; Tumor cell line growth; Culture medium; Cell proliferation; Cell culture; Serum free medium; Cell therapy; Malignant tumor; Cell line; Malignant melanoma; Tumor cell; Comparative study; Cancer
• ISSN: 0920-9069; 1573-0778
• DOI: 10.1007/s10616-010-9286-9

Melanoma cell lines are useful tools for the analysis of tumor-specific lymphocytes which are injected to patients treated by adoptive immunotherapy. So they have been established previously (with an efficacy of 47%) in Roswell Park Memorial Institute (RPMI) medium enriched with fetal calf serum (FCS). In order to improve the probability of establishing melanoma cell lines, we compared two FCS-free media with the original FCS medium. Ten melanoma-invaded lymph nodes were tested for their ability to grow in three different culture media: RPMI with FCS; RPMI with human serum (HS); serum-free X-vivo 15 (X15). For each medium, we compared the following criteria: percentage of lines obtained; period of establishment; cell morphology; expression of melanoma-associated antigens and surface molecules. More cell lines were obtained with HS and X15 media compared to FCS medium (7/10, 5/10 and 4/10, respectively). The time period to establish a stable line was similar for the three media. No morphological differences were observed in cells derived from the same tumor sample in the different media. With the X15 medium, cells generally expressed lower levels of melanocytic differentiation antigens and surface molecules. The growth of melanoma cell lines in FCS-free culture media appears possible and advantageous, with an increased probability of obtaining autologous tumor cell lines. Furthermore the cells obtained could be used as multiple antigenic sources in active or adoptive immunotherapy protocols.