人脂肪源干细胞分离培养及向成骨细胞的诱导分化

背景：脂肪源干细胞可分泌众多的免疫调节因子,不引起T细胞的细胞毒作用,并可通过调整T淋巴细胞的种类和数量。 目的：探讨人脂肪源干细胞在体外分离培养扩增的方法及向成骨细胞诱导分化的能力。 方法：以0.1%的Ⅰ型胶原酶通过组织消化的方法分离人脂肪组织中的干细胞,体外扩增培养至第2代后检测其表面抗原的表达,并在成骨诱导液中促进其向成骨细胞的分化,通过碱性磷酸酶染色、茜素红染色及对碱性磷酸酶的RT-PCR检测来明确其分化能力。 结果与结论：体外分离培养的脂肪源干细胞生长稳定,扩增速度快。流式细胞仪检测结果显示其高表达干细胞相关抗原。向成骨细胞诱导后经免疫组化染色可见矿化结节形成,RT-PCR检测发现碱性磷酸酶表达阳性。提示脂肪源干细胞在体外分离培养方法简单,扩增速度快,并具有定向分化的能力,是可靠的组织修复和细胞治疗的种子细胞来源。

Isolation,culture and osteogenic differentiation of human adipose-derived stem cells

BACKGROUND:Adipose-derived stem cells (ADSCs) can secrete a large number of immune regulatory factors, not cause T-cell cytotoxicity, and adjust the type and quantity of T lymphocytes. OBJECTIVE:To establish a method for the isolation and culture of human ADSCs and to explore the ability of osteogenic differentiation. METHODS:ADSCs were isolated from the inguinal fat pads and digested with 0.1% type I collagenase, and cultured in osteogenic medium to study the osteogenic differentiation potential by using alkaline phosphatase staining, alizarin red staining and RT-PCR. RESULTS AND CONCLUSION:ADSCs could be expanded steadily in vitro and flow cytometry revealed high expression of stem cell-associated markers. The cytoplasm of the induced cells became black by alkaline phosphatase staining, the mineralized nodule became reddish-brown by alizarin red, RT-PCR detection of alkaline phosphatase positive expression. Human ADSCs are an ideal source of adult stem cells for use in the organization repair and cell therapy.