冻存复苏后骨髓间充质干细胞体外扩增和多向分化潜能研究  

Ex vivo expansion and pluripotential differentiation of cryopreserved bone marrow mesenchymal stem cells

作　　者：项盈[1] 贾冰冰[1] 沈丹[2] 黄国平[1] 解纯刚[1] 郑强[3] 金洁[2] 王金福[1] 

机构地区：[1]浙江大学生命科学学院,浙江杭州310012 [2]浙江大学附属第一医院,浙江杭州310006 [3]浙江大学附属第二医院,浙江杭州310009

出　　处：《浙江大学学报（工学版）》2006年第11期2016-2020,共5页Journal of Zhejiang University：Engineering Science

基　　金：浙江省科技厅重点计划资助项目(2003C23015);浙江省自然科学基金资助项目(Y204139)

摘　　要：冻存复苏的人骨髓间充质干细胞(hMSCs)被传至15代并分别诱导成软骨细胞、脂肪细胞和神经细胞,以此研究冻存干细胞的扩增和分化潜能.结果表明,复苏细胞在软骨细胞诱导液的作用下分化为软骨细胞,对甲苯胺蓝异染并通过RT-PCR检测出Ⅱ型前胶原mRNA的表达;在脂肪细胞诱导液的作用下,可检测到被诱导细胞中脂滴的产生,通过油红O染色显示出细胞核周围聚集的脂滴;在神经细胞诱导液的作用下,通过免疫组化和RT-PCR均检测出被诱导细胞具有巢蛋白和神经元特异性烯醇化酶的表达,分化细胞延伸分支,末端为典型曲张体.推断复苏的hMSCs仍是具有向软骨、脂肪和神经元多系分化的潜能性干细胞群,可以作为细胞组织工程和治疗相关疾病的一种干细胞来源.Cryopreserved human bone marrow mesenchymal stem cells （hMSCs） were thawed and passaged for more than 15 generations. Resuscitated hMSCs at passage 15 were induced into chondrocytes, adipocytes and neural cells under corresponding conditioned induction medium respectively. The resuscitated cells differentiated into chondrocytes after exposure to transforming growth factor β1, insulin-like growth factor I and vitamin C. The induced cells were heterochromatic to toluidine blue and expressed cartilage matrix-procollagenal （Ⅱ） mRNA. hMSCs induced with adipogenical induction medium consisting of dexamethasone, 3-isobutyl-1-methylxanthine, indomethacin and insulin-like growth factor Ⅰ accumulated lipid vacuoles as detected by oil red O. Nestin and neuron-specific enolase （NSE）, special surface markers asso- ciated with neural cells were expressed in cells induced with a combination of β-mercaptoethanol, dimethyl sulfoxide, butylated hydroxyanisole, hydrocortisone, insulin-like growth factor Ⅰ （IGF-Ⅰ） and all-trans retinoic acid. The refractile cell bodies extended long processes that terminated at typical growth cones and filopodia. It is inferred that the resuscitated hMSCs compose a population of pluripotent cells having differentiation potential, and that cryopreserved hMSCs can be used for tissue engineering and treatment of various diseases.

关 键 词：冻存 骨髓间充质干细胞 分化 软骨细胞 脂肪细胞 神经细胞 

分 类 号：R239.28[医药卫生—中医学]