Research on the culture process and establishment of immortal cell lines from peripheral blood of newly diagnosed acute leukemia patients

Tóm tắt

Objectives: To establish a complete culture protocol for immortalizing blast cells from newly diagnosed acute leukemia (AL) patients, thereby creating a personalized in vitro model. Subjects and Methods: Peripheral blood samples were collected from thirty newly diagnosed AL patients. Mononuclear cells (MNCs) were isolated and cultured in various media: RPMI 1640 + 10% fetal bovine serum (FBS) + 1x penicillin/streptomycin (P/S), RPMI + 20% FBS + 1x P/S, RPMI + 5% fetal human serum (FHS) + 1x P/S, and StemMACS™ HSC-CFU complete with Epo + 1x P/S. Cell viability, proliferation were evaluated periodically over three months. Results: The immortalization culture protocol was successfully established using two media: RPMI + 20% FBS and RPMI + 5% FHS. The immortal cell lines from these two media showed a cell doubling time of 48–72 hours. In the StemMACS matrix gel, all cell lines differentiated, while in the 10% FBS medium, two out of three cell lines died after two weeks of culture. The immortalized cells maintained their typical blast morphology and were able to proliferate stably for over 30 passages. Conclusion: The study successfully established a culture protocol for 3 immortal cell lines from the 30 blood samples of AL patients. This represents a significant breakthrough in blood cancer research in Vietnam, providing a valuable research model for drug screening and the development of personalized treatment strategies, thereby improving treatment effectiveness and patient prognosis.