The study was aimed to evaluate if the modified in situ cryopreservation could affect the biological function of mesenchymal stem cells (MSC) in vitro. Mesenchymal stem cells from human bone marrow were isolated by standard method and characterized with their morphology, cell-surface antigen profile and differentiation repertoire in vitro. The culture-expanded MSC were cryopreserved in situ with culture medium (DMEM-LG) containing 10% D MSO and 30% selected FCS in -70 degrees C. Following recovery of cryopreservation, differentiation to adipocytes, chondrocytes, and osteoblast in vitro and cell cycle analysis were performed to investigate whether the cryopreservation would change the differentiation potential of MSC. The results showed that after recovery of cryopreservation, there was no changes detected as compared with the culture-expanded MSC in both differentiation potency and growth pattern at 12 weeks. In conclusions: this optimized short term in situ cryopreservation at -70 degrees C could retain biological characteristics of human MSC for at least 3 months, and this method may be useful for cryopreservation of hum an bone marrow MSCs.
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