经血源子宫内膜干细胞培养、鉴定及体外分化潜能的研究

Isolated Culture, Identifi cation and Differentiation Potential of Menstrual Blood-derived Mesenchymal Stem Cells

现阶段干细胞的来源常具有侵入性,该文旨在研究新来源于经血的经血源子宫内膜干细胞(menstrual blood-derived mesenchymal stem cells,MenSCs)的基本生物学特性及分化潜能。采用密度梯度法从女性经血中分离MenSCs,测定MenSCs群体倍增时间,流式细胞仪鉴定细胞表面抗原,免疫荧光法检测MenSCs nestin阳性表达情况,体外验证其成骨成脂分化潜能。结果表明,MenSCs具有典型的梭状结构,细胞倍增时间为32.2 h,均一地高表达CD29、CD90及CD105,不表达CD14、CD45、HLA-DR。免疫荧光表明,MenSCs为nestin阳性。MenSCs成脂诱导后,油红O染色为阳性。成骨诱导前期诱导组细胞胶原表达量升高,诱导两周后MenSCs形成钙结节,诱导组细胞ALP(alkaline phosphatase)活性连续3周呈上升趋势。以上证明,MenSCs具有来源广泛的优势,具有较高的增殖能力、较低免疫原性、nestin阳性及多向分化潜能等特性,可成为干细胞治疗的理想种子细胞。

Stem cell therapy can help cure many diseases. Yet many of the suitable cells currently identified for human use are difficult to obtain and involve invasive procedures. In this research, we investigated the basic biological characteristics and differentiation potential of novel menstrual blood-derived mesenchymal stem cells （MenSCs）. MenSCs specific markers were analyzed by a flow cytometer. Nestin expression was evaluated with immunofluorescence. The capacity of osteoblast and adipocyte to differentiate was evaluated. Our results showed that MenSCs were successfully isolated from female menstrual blood. The doubling rate of MenSCs was 32.2 h. MenSCs were positive for CD29, CD90, CD105, while negative for CD14, CD45, HLA-DR. Immunofluorescence showed that MenSCs exhibited nestin-positive characteristics. MenSCs were positive for Oil Red O staining after inducing by adipogenic differentiation medium. After exposure in osteogenetic differentiation media, collagen expression of MenSCs was higher at the early age and Alizarin Red staining evidenced that MenSCs exhibited osteocyte phenotypes after 2 weeks. In addition, ALP activity analysis illustrated that MenSCs was on the rise for3 weeks. These results indicate that MenSCs have properties of low immunogenicity, high proliferation potential and multi-linage differentiation capacity. Our study suggests that MenSCs might be an ideal resource for stem cell therapy and regenerative medicine applications.