崂山奶山羊脂肪间充质干细胞系的建立及鉴定

旨在建立崂山奶山羊脂肪间充质干细胞系并对其进行初步鉴定.采集崂山奶山羊脂肪组织,在Ⅰ型胶原酶的分解消化作用下,将脂肪组织中的单核细胞进行分离并扩增培养;测定其生长曲线,利用Real time RT-PCR技术对其表达的干细胞因子进行鉴定;将传至第3代的脂肪间充质干细胞进行成骨诱导分化,利用茜素红染色进行鉴定.结果显示,崂山奶山羊脂肪间充质干细胞形态为长梭形、星形等成纤维细胞样细胞形态,但比成纤维细胞饱满,第3代细胞生长至3 d左右时细胞进入指数生长期,生长至6~7 d时进入平台期;经成骨诱导分化后,经茜素红染色可见骨结节被染成深红色.综上提示,崂山奶山羊脂肪间充质干细胞具有诱导分化潜能.

Establishment and Identification of Adipose Mesenchymal Stem Cell Lines of Laoshan Dairy Goat

The aim of this study was to establish adipose mesenchymal stem cell lines of Laoshan dairy goats and to identify them.The adipose tissues of Laoshan dairy goats were collected,and the monocytes in the adipose tissues were isolated and cultured under the decomposition and digestion of typeⅠcollagenase.The growth curve was determined and the expression of stem cell factors was identified by Real time RT-PCR.The third-generation adipose mesenchymal stem cells were induced by osteogenesis and identified by alizarin red staining.The results showed that the adipose mesenchymal stem cells of Laoshan dairy goat were long fusiform and stellate fibroblast-like cells,but were fuller than the fibroblasts.When the third-generation cells grew to 3 days or so,the cells entered the exponential growth phase and 6-7 days entered the plateau stage.After osteogenic induction,alizarin red staining showed that the bone nodules were stained dark red.The results demonstrated that adipose mesenchymal stem cells of Laoshan dairy goat had the potential to induce differentiation.