人脐血间充质干细胞的生物学特性及其成骨成脂肪分化(英文)  

Biological characteristics,osteogenic and adipogenic differentiation of mesenchymal stem cells from human umbilical cord blood

作　　者：张才龙[1] 那娜[2] 张积华[3] 孙康[1] 田少奇[1] 夏长所[1] 

机构地区：[1]青岛大学医学院附属医院关节外科,266003 [2]青岛市第八人民医院妇产科 [3]青岛大学医学院附属医院健康查体中心,266003

出　　处：《中国组织工程研究》2012年第10期1773-1779,共7页Chinese Journal of Tissue Engineering Research

基　　金：山东省科技攻关计划(2008GG30002037),课题名称:脐血间充质干细胞移植修复骨坏死的实验研究~~

摘　　要：背景:目前有关脐血间充质干细胞的生物学特性及分化能力的研究较少。目的:观察人脐血间充质干细胞的生物学特性,及其向成骨、成脂肪细胞分化的能力。方法:从不同胎龄脐血中分离间充质干细胞,对其进行原代和传代培养,并诱导其向成骨及成脂肪细胞分化。结果与结论:倒置相差显微镜下见分离培养的脐血间充质干细胞贴壁生长,呈成纤维细胞样外观,细胞呈螺旋状排列;透射电镜下可见脐血间充质干细胞胞核比例大,细胞器少,为低分化细胞;原代及传代培养的脐血间充质干细胞生长曲线均呈S型,第3,5代细胞增殖能力最强,低胎龄的脐血间充质干细胞集落形成能力最强。流式细胞仪检测结果显示,脐血间充质干细胞稳定表达间充质干细胞相关抗原CD29,CD44和CD90,不表达造血细胞标志CD34和CD45。成骨诱导后3周,碱性磷酸酶染色为强阳性,茜素红染色可见大量钙化基质的形成;成脂诱导3周,油红O染色可检测到胞质中脂滴的形成。提示脐血间充质干细胞具有间充质干细胞的形态特征、生长增殖特点及细胞表面标志物等生物学特性,可向成骨细胞及脂肪细胞分化。BACKGROUND：Up to now,there are few reports addressing the biological properties and differentiation potential of umbilical cord blood-derived mesenchymal stem cells（UCB-MSCs）.OBJECTIVE：To observe the biological characteristics,osteogenic and adipogenic differentiation potential of UCB-MSCs.METHODS：MSCs were harvested and cultured from UCB at various gestational ages（GA）.Harvested UCB-MSCs were cultured primarily and subcultured,and then induced to differentiate into osteoblasts and adipocytes.RESULTS AND CONCLUSION：Under the inverted phase contrast microscope,UCB-MSCs adhered to the wall,showing fibroblast like morphology and whirlpool like growth alignment.Observation of the ultramicrostructure under transmission electron microscope showed that UCB-MSCs had a big cell nucleus,fewer cellular organelles and big karyoplasmic ratio.All of the growth curves of primary and passaged UCB-MSCs presented S-shaped.The 3rd and 5th passages of MSCs showed the strongest proliferation activity.The count of colony forming unit fibroblasts varied with GA,significant difference was found among the three GA groups（P 0.05）,and the lower GA group had a higher count of colony forming unit fibroblasts than that in the older GA group.Flow cytometry showed that these cells expressed CD29,CD44 and CD90 positively,but they failed to express hematopoiesis related molecules such as CD34 and CD45.When the MSCs were induced to osteogenic and adipogenic differentiation for 3 weeks,strong expression of alkaline phosphatase was found and the formation of a mineral extracellular matrix was detected by alizarin red staining were detected;and neutral lipid vacuoles were detected by oil red O staining.UCB-MSCs have similar morphological and biological characteristics and cell surface molecule markers with MSCs derived from bone marrow,both of which have great capability of proliferation and regeneration.UCB-MSCs can be induced to osteoblasts and adipocytes in a suitable condition in vitro.

关 键 词：脐血 间充质干细胞 生物学特性 成骨细胞 成脂肪细胞 

分 类 号：R394.2[医药卫生—医学遗传学]