中华显微外科杂志
中華顯微外科雜誌
중화현미외과잡지
Chinese Journal of Microsurgery
2009年
3期
210-212,插6
,共4页
周佳%沈尊理%金羽青%刘伟%沈华
週佳%瀋尊理%金羽青%劉偉%瀋華
주가%침존리%금우청%류위%침화
间充质干细胞%脐带%软骨%显微外科
間充質榦細胞%臍帶%軟骨%顯微外科
간충질간세포%제대%연골%현미외과
Mesenehymal stem cells%Umbilical cord%Chondrocytes%Microsurgery
目的 研究脐带间充质干细胞获得纯化的高效方法及向软骨细胞的分化能力.方法 取人的正常分娩或剖腹产胎儿的脐带,用胶原酶和胰酶初步消化后,将流式细胞仪筛选得到的CD45(-)、CD90(+)细胞作为原代细胞进行培养,并用复合胶原酶差速消化法逐级传代、纯化细胞,每次传代后取部分细胞用作流式细胞仪分析细胞的CD45、CD90阳性比率;对P3代细胞向软骨细胞进行诱导.结果 流式细胞仪检测提示,采用流式细胞仪进行初次筛选并用复合胶原酶差速消化法逐级传代获得的P1、P2、P3代细胞中CD90的阳性率为(80.86±7.85)%、(95.86±3.28)%、(97.15±1.43)%,而CD45的阳性率为(2.53±0.28)%、(0.97±0.48)%、(0.05±0.01)%;向软骨细胞诱导结果提示P3代细胞有向终末软骨细胞分化能力,具有干细胞的特性.结论 采用流式细胞仪进行初次筛选并用复合胶原酶差速消化法能快速获得高纯度的具有向终末软骨细胞分化能力的人脐带间充质干细胞.
目的 研究臍帶間充質榦細胞穫得純化的高效方法及嚮軟骨細胞的分化能力.方法 取人的正常分娩或剖腹產胎兒的臍帶,用膠原酶和胰酶初步消化後,將流式細胞儀篩選得到的CD45(-)、CD90(+)細胞作為原代細胞進行培養,併用複閤膠原酶差速消化法逐級傳代、純化細胞,每次傳代後取部分細胞用作流式細胞儀分析細胞的CD45、CD90暘性比率;對P3代細胞嚮軟骨細胞進行誘導.結果 流式細胞儀檢測提示,採用流式細胞儀進行初次篩選併用複閤膠原酶差速消化法逐級傳代穫得的P1、P2、P3代細胞中CD90的暘性率為(80.86±7.85)%、(95.86±3.28)%、(97.15±1.43)%,而CD45的暘性率為(2.53±0.28)%、(0.97±0.48)%、(0.05±0.01)%;嚮軟骨細胞誘導結果提示P3代細胞有嚮終末軟骨細胞分化能力,具有榦細胞的特性.結論 採用流式細胞儀進行初次篩選併用複閤膠原酶差速消化法能快速穫得高純度的具有嚮終末軟骨細胞分化能力的人臍帶間充質榦細胞.
목적 연구제대간충질간세포획득순화적고효방법급향연골세포적분화능력.방법 취인적정상분면혹부복산태인적제대,용효원매화이매초보소화후,장류식세포의사선득도적CD45(-)、CD90(+)세포작위원대세포진행배양,병용복합효원매차속소화법축급전대、순화세포,매차전대후취부분세포용작류식세포의분석세포적CD45、CD90양성비솔;대P3대세포향연골세포진행유도.결과 류식세포의검측제시,채용류식세포의진행초차사선병용복합효원매차속소화법축급전대획득적P1、P2、P3대세포중CD90적양성솔위(80.86±7.85)%、(95.86±3.28)%、(97.15±1.43)%,이CD45적양성솔위(2.53±0.28)%、(0.97±0.48)%、(0.05±0.01)%;향연골세포유도결과제시P3대세포유향종말연골세포분화능력,구유간세포적특성.결론 채용류식세포의진행초차사선병용복합효원매차속소화법능쾌속획득고순도적구유향종말연골세포분화능력적인제대간충질간세포.
Objective To explore the efficient way to purify and to enrich umbilical cord mesenchymal stem cells which have abilities to differentiate into chondrocytes. Methods Umbilical cord was acquired from new born infants and chopped into 1 mm2 cubics, then they were pretreated with colleganase and trypsin so as to get the mixed cell suspension. CD45(-),CD90(+) cells were acquired from the mixed cell suspension via FCM sorting and were cultured in the medium as umbilical cord mesenchymal stem PO cells. The PO cells were detached and passaged by multiplex collagenase NB4 and the positive rate of CD90 and CD45 of the P1-P3 cells were observeed via FCM ; the P3 cells were induced to differentiated into chondrocytes. Results FCM results showed that the positive rate of CD90 on P1, P2, F3 was (80.86 ± 7.85)%,(95.86± 3.28)%,(97.15± 1.43)%, while the positive rate of CD45 on P1, P2, P3 cells was (2.53 ±0.28)% and (0.97 ± 0.48)% and (0.05 ± 0.01)% respectively; P3 cells had differenciated into chondrocytes. Conclusion FCM presorting and Multiplex collagenase detachment procedures can swiftly acquire highly purified human umbilical cord mesenchymal stem cells which have the ability to differentiate into chondroeytes.
