| 摘要: |
| 目的:对新鲜和超低温保存的骨髓来源单个核细胞(MNC)体外扩增的内皮祖细胞(EPC)进行功能比较。方法:从猪髂骨抽取骨髓,对分离后的MNC进行培养或-80℃冻存3个月后再培养；冻存后培养的P1代细胞利用免疫组化及流式细胞技术进行EPC表面标志抗原鉴定。同时分别对新鲜和冻存培养的EPC获得率、细胞迁徙、黏附和增殖功能进行比较。结果:冻存组细胞免疫组化鉴定： CD133(+)、CD34(+)、CD31()、KDR(),流式细胞技术鉴定：CD133的阳性率(17.24±3.12)%,CD34的阳性率(37.21±10.85) %,CD31的阳性率(72.07±13.34) %,KDR的阳性率(89.09±16.40)%。新鲜和冻存的MNC经诱导培养后EPC获得率分别为(1.1±0.078)%、(1.03±0.061)%, P=0.054；细胞迁徙率分别为(15±0.71)%、(14.2±0.63)%, P=0.17;贴壁率分别为(42.7±2.1)%、(39.5±1.7)%, P=0.11; 增殖功能分别为 (25.06±2.82)×104、(21.64±2.34)×104, P=0.089。结论:超低温保存骨髓来源的MNC经诱导培养得到的EPC其数量和功能均无明显影响,该方法可在短时间得到大量的同质EPC,为EPC移植提供了来源保障。 |
| 关键词: 内皮祖细胞 冻存 单个核细胞 细胞分化 |
| DOI：10.3724/SP.J.1008.2009.01225 |
| 投稿时间：2009-01-06修订日期：2009-10-21 |
| 基金项目:国家自然科学基金 (30672170). |
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| Differentiation of cryopreserved bone marrow-derived mononuclear cells into endothelial progenitor cells |
| WU Jian-guo1, LUO Tian-hang1, ZHOU Hong2, XUE Xu-chao1, BI Jian-wei1,FANG Guo-en1* |
| (1.Department of General Surgery, Changhai Hospital,Second Military Medical University,Shanghai 200433,China;2.Experimental Center of Department of Hematology,Changhai Hospital,Second Military Medical University,Shanghai 200433) |
| Abstract: |
| Objective:To compare the functions of endothelial progenitor cells (EPCs) differentiated from cryopreserved and fresh bone marrow-derived mononuclear cells (MNCs). Methods: The bone marrow samples were taken from swine iliac bones. The isolated MNCs were cultured or cryopreserved at -80℃ for 3 months and then cultured again. The P1-EPCs were identifed by Dil-ac-LDL and FITC-UEA-1 double staining,immunohistochemistry and flow cytometry. The EPC pick-up rate,migration,adhesion, and proliferation abilities were compared between the cryopreserved group and the fresh group. Results: Immunohistochemisty showed that the P1-EPCs of the cryopreserved group were positive for CD133 (+),CD34 (+),CD31 () and KDR (); flow cytometry also showed they were positive for CD133 (\[17.24±3.12\]%),CD34 (\[37.21±10.85\]%),CD31 (\[72.07±13.34\]%) and KDR (\[89.09±16.40\]%). There were no significant differences in the pick-up rates (\[1.1±0.078\]% vs \[1.03±0.061\]%,P=0.054),migration rates (\[15±0.71\]% vs \[14.2±0.63\]%,P=0.17),adherence rates (\[42.7±2.1\]% vs \[39.5±1.7\]%,P=0.11),and proliferation abilities (\[25.06±2.82\]×104 vs \[21.64±2.34\]×104,P=0.089) between EPCs of the fresh and cryopreserved groups. Conclusion: Cryopreservation has no measurable influence on the numbers and functions of EPCs differentiated from bone marrow-derived MNCs,so cryopreservation can be used to obtain sufficient homogeneous EPCs in a short period for therapy using EPCs transplantation. |
| Key words: endothelial progenitor cells cryopreservation mononuclear cells cell differentiation |
