摘要
目的研究作为核移植供体细胞的人骨髓基质细胞相关生物学特性。方法应用密度梯度离心法分离培养人骨髓基质细胞，并对其进行形态观察、免疫荧光分析细胞骨架结构、端粒酶活性测定、核型及细胞周期分析。结果培养的细胞具有正常的大小、形态、细胞骨架结构：在融合密度达80%-90%时，G0＋G1期细胞所占的比例较高，约占92．58％；检测传至第7代的人骨髓基质细胞具有正常染色体数目（46，XY），端粒酶活性为0．567。结论形态良好、染色体数目正常的人骨髓基质细胞可以作为核移植的供体细胞。
Objective To research the related biological characteristics of human bone marrow stromal cells （hBMSCs） as donor cells during nuclear transfer. Methods hBMSCs were isolated by density gradient centrifugation and cultured in vitro at 37 ℃, 5% CO2. Morphological observations were performed with phase contrast microscope; cytoskeleton of hBMSCs was detected by immunofluorescence; telomerase activities were determined by PCR-ELISA; karyotype and cell cycle of the hBMSCs were respectively detected by Giemsa staining and flow cytometry. Results All collected by the density gradient separation method showed the similar appearance, displaying long spindle shaped. The cultured hBMSCs possessed the normal cell size, morphology and cytoskeleton. The cell cycle assay showed that G0＋G1 phage was in the high percentage （about 92.58%）. The hBMSCs at the 7th passage had normal human chromosome number （46, XY）, and their telomerase activity was 0.567. Conclusion hBMSCs with fine morphology and normal chromosome number can be considered as the donor cells during nuclear transfer.
出处
《中华神经医学杂志》
CAS
CSCD
2007年第3期221-224,共4页
Chinese Journal of Neuromedicine
基金
国家自然科学基金（30270491）
