Abstract:

BACKGROUND: The proliferation of human umbilical cord mesenchymal stem cells (hUCMSCs) strongly depends on the culture conditions. Medium containing 10-20% fetal bovine serum (FBS) can promote cell growth.
OBJECTIVE: To build the serum-decremental technology for UCMSCs culture.
METHODS: hUCMSCs were separated using collagenase Ⅱ digestion method and purified by adherent culture. The cells were cultured and expanded by the serum-decremental method: media 20% serum-free medium for the primary culture, 50% serum-free medium for the second passage, 80% serum-free medium for the third passage, and 100% serum-free medium for the fourth passage. Cells were cultured in α-MEM medium containing 10% FBS as control. Cell surface markers were tested by flow cytometry. Osteogenic induction test was performed.
RESULTS AND CONCLUSION: Cells cultured in the both media were similar in proliferative ability, morphology, and immunophenotype. Their differentiative potential kept well in both media. However, less FBS was used in serum-decremental medium. Serum-decremental medium improves the safety of hUCMSCs culture in clinical practice.