冻存前后骨髓间充质干细胞的生物学特性和支持造血的研究.docVIP

冻存前后骨髓间充质干细胞的生物学特性和支持造血的研究 nbsp;nbsp;nbsp;nbsp;nbsp;nbsp;nbsp;nbsp;nbsp; 作者：赵智刚，唐晓琼，黎纬明，游泳，邹萍 【摘要】nbsp; 为了了解冻存复苏过程对骨髓间充质干细胞 (mesenchymal stem cell，MSC) 生物学特性和支持造血能力的影响，采用常规方法分离培养骨髓MSC，将传代后的细胞用含10％二甲亚砜、40％胎牛血清的IMDM细胞冻存液保存在-196℃液氮中，观察短期（4周）和中期（9-15月）复苏后MSC的活性、免疫表型、多向分化能力和支持造血的能力，并同冻存前的MSC进行比较。 结果表明：中短期冻存后的MSC的细胞活性分别为(90士3.75)％和(93士2.51)％；冻存后细胞的增殖能力、免疫表型、体外分化为脂肪和骨细胞的能力、支持集落（CFU-GM，CFU-E，CFU-GEMM）的生长作用和冻存前MSC相似。结论：骨髓MSC在液氮中短期和中期保存后，细胞活性略有下降，但是并不影响MSC的增殖、分化和支持造血能力。 【关键词】nbsp; 支持造血 　　Biologicalnbsp;nbsp; Characteristicsnbsp;nbsp; andnbsp;nbsp; Ability of Supporting Hematopoiesis of Bone Marrow Mesenchymalnbsp;nbsp;nbsp; Stem Cells Pre-and Post-Cryop-reservation 　　Abstractnbsp; This study was aimed to observe the biological characteristics of cryopreserved bone marrow mesenchymal stem cell (MSC) and to examine theirnbsp; abilities tonbsp; support in vitronbsp; hematopoiesis.Bone marrow MSC were cryopreserved in -196℃ liquid nitrogen for 4 weeks (short term) and 9-15 months (medium term) with IMDM containing 10% DMSO，40% fetal calf serum as cryoprotectant.The viability，proliferation，immunophenotype，in vitro differentiationnbsp; and ability of supportingnbsp; hematopoiesisnbsp; of thawed MSC were investigatednbsp; and compared with these of pre-cryopreserved MSC.The results showed that the cell viability werenbsp; (93士2.51)％ and (90士3.75)％ for MSC cryopreserved as long as 4 weeks or 9-15 months respectively.However，there were no changes detected，as compared with pre-cryopreserved MSC innbsp; immunophenotype，abilities of proliferation and supporting colony forming of CFU-GM，CFU-E and CFU-GEMM.It is concluded that bone marrow-derived MSC can be stored in liquid nitrogen fornbsp; short-termnbsp; (4 weeks) or medium-term (9-15 months) without changes of abilities of proliferation，differentiation and hematopoiesis support. 　　Key words cryopreservation;nbsp; mesenchymal stem cell； hematopoiesis support 　　间充质干细胞（mesenchymal stem cell，MSC）具有多向分化