恒河猴骨髓间质干细胞的体外分离培养及形态学特征分析(英文)
Morphological Properties of Mesenchymal Stem Cells Derived from Bone Marrow of Rhesus Monkeys (in English)
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摘要: 探索恒河猴骨髓间质干细胞（MSC）的体外分离培养方法，为其应用提供实验基础。取恒河猴骨髓细胞悬液，经梯度离心去除大部分血细胞，取含有MSC的中间单核细胞层，在含10%胎牛血清及1 ng/mL碱性成纤维细胞生长因子的L-DMEM中培养扩增，并不断换液去除杂细胞，经过18 d的原代培养，获得呈致密单层生长的MSC，其形态为较规则的长梭形细胞，排列有方向性，呈现一定的漩涡状、辐射状生长趋势。将原代细胞以1∶2传代，传代培养后期，细胞增殖速度逐渐变缓，细胞形态逐渐出现三角形、多边形及扁平宽大形等不规则形态。结果显示，恒河猴骨髓间质干细胞可在体外进行传代培养，但需进一步优化其培养条件。Abstract: To establish an in vitro system for isolating and culturing the mesenchymal stem cells (MSC) of Rhesus monkeys, and to provide research data for its further application, the bone marrow of Rhesus monkeys was collected and separated by gradient centrifugation to discard most of the blood cells. The MSC contained in the monocyte centrifuging layer was obtained and cultured in Dulbecco's modified media (low glucose, L-DMEM) supplemented with 10% Fetal bovine serum (FBS) and 1 ng/ml basic fibroblast growth factor (bFGF). The non-MSC was screened out by continuously renewing the medium. A passage culture was undertaken while the MSC monolayer formed. The spindle-shaped MSC formed a monolayer after 18 days of primary culturing, and the cells appeared in an oriented array with a swirling and irradiating growth trend. In the anaphase of passage culture, the cell proliferation rate was decreased and the morphology changed into triangular, polygon and flat appearance. These results suggested that mesenchymal stem cells (MSC) of the Rhesus monkey can be passaged in vitro with the established optimized culture system.
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