人脐带间充质干细胞的体外分化及对犬外周血淋巴细胞增殖的影响

目的 通过体外实验检测人脐带间充质干细胞的体外多向分化及对犬T淋巴细胞增殖的影响,探讨其作为组织工程种子细胞的可能性.方法 体外培养人脐带间充质干细胞,分别使用含有地塞米松、转化生长因子、3-异丁基-1-甲基黄嘌呤的培养基诱导其成骨、成软骨、成脂肪三系分化后进行染色鉴定,并将经丝裂霉素处理的人脐带间充质干细胞与植物凝集素刺激下的犬外周血T淋巴细胞共培养,5d后多功能酶标仪检测吸光度值.结果 人脐带间充质干细胞体外生长良好,呈长梭形,形态均一,使用成骨、成软骨和成脂培养基分别诱导分化后碱性磷酸酶（ALP）染色、骨钙素免疫细胞化学染色、茜素红染色、亚甲基蓝染色和油红0染色分别呈阳性,人脐带间充质干细胞与犬外周血T淋巴细胞混合组吸光度值较单纯犬外周血T淋巴细胞组吸光度值明显降低.结论 人脐带间充质干细胞具有多向分化能力,并能够抑制犬外周血淋巴细胞的增殖,有望成为创伤组织及骨组织修复的种子细胞.

Differentiation of human umbilical cord mesenchymai stem cells in vitro and their effects on canine peripheral lymphocytes proliferation

Objective To explore the multiple differentiation and immune regulation on canine peripheral blood T lymphocytes in vitro of human umbilical cord mesenchymal stromal cells （UCMSCs）,so as to study their feasibility and application value of being as seed cells of tissue engineering.Methods USMSCs were cultured in vitro and induced to differentiate into osteocytes,chondrocytes and adipocytes by using the media containing dexamethasone,transforming growth factor and 3-isobutyl-1-methylxanthine,T lymphocytes from canine peripheral blood were stimulated by phytohemagglutinin and co-cultured with mitomycin-inhibited UCMSCs and a value were measured following 5 days using multifunctional microplate reader.Results UCMSCs proliferated actively and exhibited spindle-shaped morphology in vitro,Alkaline phosphatase （ALP） staining,Osteocalcin （OCN） immunocytochemical staining,Alizarin red staining,Alcian blue staining and Oil red O staining were positive,the A value were significantly lower in the co-culture of UCMSCs and T lymphocytes group than T lymphocytes alone group.Conclusions UCMSCs have multiple differentiation properties,and could inhibit the proliferation of canine peripheral blood T lymphocytes,they are optimal cell source for tissue engineering.