In vitro culture of human acute lymphoblastic leukemia (ALL) cells in serum-free media; a comparison of native ALL blasts, ALL cell lines and virus-transformed B cell lines.
The aim of this study was to standardize in vitro culture conditions for human acute lymphoblastic leukemia (ALL) cells. The cells were cultured in medium containing 10% fetal calf serum (FCS) and in the four serum-free media X-vivo 10, X-vivo 15, X-vivo 20 and Stem Span. Native ALL blasts could proliferate in all four serum-free media, but the strongest responses were usually observed with Stem Span. Native leukemia blasts were also cultured in the presence of various single cytokines or cytokine combinations. The highest proliferation was usually observed in the presence of Flt3-Ligand (Flt3-L) when single cytokines were examined, and these responses could be further increased especially by combining Flt3-L with interleukin 3 (IL3), IL7 or stem cell factor (SCF). Proliferation could also be increased when ALL blasts were cultured in the presence of two commercially available fibroblast cell lines (Hs27 and HFL1). Based on these results we suggest that in vitro culture conditions for native human ALL blasts can be standardized by using serum-free culture media supplemented with exogenous Flt3-L+IL3+SCF, and the use of accessory cells can also be standardized by using well-characterized fibroblast cell lines. Detectable ALL blast proliferation can then be observed for most patients. Our experimental model can thereby be used for in vitro evaluation of possible antileukemic treatment strategies, and it will then allow comparison of experimental results between different studies.
Bruserud
Øystein
Ø
Medical Department, Division of Hematology, Institute of Medicine, Haukeland University Hospital, N-5021 Bergen, Norway. oystein.bruserud@haukeland.no
Glenjen
Nils
N
Ryningen
Anita
A
Ulvestad
Elling
E
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001402:B-Lymphocytes / Q000166:cytology* / Q000187:drug effects; D002051:Burkitt Lymphoma / Q000473:pathology; D002461:Cell Line, Transformed / Q000166:cytology / Q000187:drug effects; D018920:Coculture Techniques; D003470:Culture Media / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D046508:Culture Techniques / Q000379:methods*; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D007378:Interleukins / Q000494:pharmacology; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000473:pathology; D008297:Male; D008565:Membrane Proteins / Q000494:pharmacology; D008875:Middle Aged; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000187:drug effects; D015452:Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / Q000473:pathology; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000473:pathology*; D019089:Stem Cell Factor / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000166:cytology* / Q000187:drug effects
D003470:Culture Media; D016895:Culture Media, Serum-Free; D007378:Interleukins; D008565:Membrane Proteins; D019089:Stem Cell Factor; C084881:flt3 ligand protein