冻存骨髓来源单个核细胞分化成内皮祖细胞的功能特性

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...

Differentiation of cryopreserved bone marrow-derived mononuclear cells into endothelial progenitor cells

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 Pl-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 Pl-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] X 10~4 vs [21. 64± 2. 34]×10~4 ,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.