同步诱导培养兔骨髓源性内皮祖细胞和平滑肌祖细胞

发布时间：2018-05-08 09:41

本文选题：内皮祖细胞 + 平滑肌祖细胞　；参考：《第二军医大学学报》2014年01期

【摘要】：目的采用兔骨髓来源的单个核细胞同步诱导分化为兔内皮祖细胞(EPCs)和平滑肌祖细胞(SPCs),研究其生物学特性,评估其作为组织工程化静脉瓣种子细胞的可能性。方法梯度密度离心法获取兔骨髓血单个核细胞沉淀,分别用含5%胎牛血清(FBS)的EGM-2完全培养液向EPCs方向诱导培养;用含20ng/mL血小板源性生长因子BB、5%FBS,不含血管内皮生长因子(VEGF)的EBM-2培养液向SPCs方向诱导培养。鉴定两种细胞的分化情况。结果诱导的EPCs培养至10d左右,细胞单层融合呈"铺路石"状;表达血管内皮细胞生长因子受体2(VEGFR-2)、血管性血友病因子(vWF)、CD133,不表达α-平滑肌肌动蛋白(α-SMA);透射电镜可见细胞质内特征性Weibel-Palade小体;细胞生物学功能检测可见EPCs在基质胶上呈现血管状生长。诱导的SPCs培养至14d左右呈现血管平滑肌细胞"峰-谷"样生长特性;表达CD34、α-SMA,不表达vWF和VEGFR-2;在透射电镜下可见细胞内含有与细胞纵轴平行排列的肌丝;在基质胶上不规则生长。结论兔骨髓血梯度密度离心得到的单个核细胞可同步诱导分化为EPCs和SPCs,为构建组织工程化静脉瓣提供了经济且可简便获取的种子细胞。
[Abstract]:Objective to study the biological characteristics of rabbit endothelial progenitor cells (EPCs) and smooth muscle progenitor cells (SPCs1) induced by mononuclear cells derived from rabbit bone marrow, and to evaluate their potential as seed cells for tissue engineering venous valve. Methods Rabbit bone marrow mononuclear cells were precipitated by gradient density centrifugation and cultured in the direction of EPCs by EGM-2 complete culture medium containing 5% fetal bovine serum. EBM-2 medium containing 20ng/mL platelet-derived growth factor BB, 5S and no vascular endothelial growth factor (VEGF) was used to induce the culture in the direction of SPCs. The differentiation of two kinds of cells was identified. Results the monolayer fusion of EPCs was "paving stone" when cultured for 10 days or so. The expression of vascular endothelial growth factor receptor (VEGFR-2), von Willebrand factor (VWFU) CD133and 伪 -SMAA (伪 -SMAA) were not detected. The characteristic Weibel-Palade bodies in cytoplasm were observed under transmission electron microscope. Cell biological function test showed that EPCs showed vascular growth on matrix glue. After cultured for 14 days, SPCs showed a "peak-valley" growth characteristic, expression of CD34, 伪 -SMA, no vWF and VEGFR-2. Under transmission electron microscope, the cells had myofilaments parallel to the longitudinal axis of the cells and grew irregularly on the matrix glue. Conclusion the mononuclear cells obtained from rabbit bone marrow blood gradient density centrifugation can be induced to differentiate into EPCs and SPCs simultaneously, which provides an economical and easy to obtain seed cells for the construction of tissue engineered venous flap.
【作者单位】：泰山学院体育学院理论教研室;第二军医大学基础部人体解剖与组织胚胎学教研室;泰安市妇幼保健院儿童保健科;
【基金】：国家自然科学基金(30672045)~~
【分类号】：R318.08

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