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目的:从早期人胚分离培养人早期胚胎间充质干细胞(mesenchymal stem cell,MSC),并初步鉴定其生物学特性。方法:取4-6周胚龄的人胚,用免疫组织化学法,结合特定抗体SH-2,对MSC在人胚中的分布进行定位,分离组织进行培养。免疫组化和流式细胞术检测MSC的相对特异性抗原SH-2、CD44、ODT-4、S-100、CD34、α-smooth-actin在分离培养细胞中的表达。结果:在胚胎的四肢、躯体部紧邻神经管的下方(不包括原肠等内脏部位)有大量的MSC:分布。分离培养可得到成纤维状的MSC,贴壁生长,有生长优势,可多次传代并保持MSC:生物学特性不变。结论:通过机械分离和胰酶消化的方法,可从早期人胚得到MSC。利用MSC的贴壁生长特性及生长优势,经4-5次传代后可得到纯度很高的MSC。
OBJECTIVE: To isolate and culture human early embryonic mesenchymal stem cells (MSC) from early human embryos and to identify their biological characteristics. Methods: Human embryos of 4-6 weeks embryo age were harvested, and the distribution of MSC in human embryos was localized by immunohistochemistry and specific antibody SH-2. The tissues were isolated and cultured. The expression of SH-2, CD44, ODT-4, S-100, CD34 and α-smooth-actin in MSC were detected by immunohistochemistry and flow cytometry. RESULTS: There were a large number of MSC: distributions in the limbs of the embryo, just below the neural tube of the body (excluding internal organs such as gastrula). Isolation and culture of fibroblasts can be obtained MSC, adherent growth, growth and can be multiple passages and maintain MSC: biological characteristics unchanged. Conclusion: MSCs can be obtained from early human embryos by means of mechanical separation and trypsinization. Adopting the adherent growth characteristics and growth advantages of MSC, the highly pure MSCs can be obtained after 4-5 passages.