CAJ | 학술논문

目的观察体外培养大鼠骨髓间充质干细胞（MSCs）的生物学特征并研究DAPI和GFP双重标记的MSCs向移植肺的趋化现象。方法全骨髓贴壁培养法分离培养大鼠MSCs ，流式细胞法检测细胞表型。观察包括细胞形态学、生长曲线在内的生物学特征。通过定向诱导MSCs向成骨和成脂肪分化鉴定其多向分化潜能。绿色荧光蛋白（GFP）和4＇，6-二脒基-2-苯基吲哚（DAPI）双重标记MSCs ，经尾静脉注入受体大鼠，取肺组织冰冻切片，观察MSCs是否趋化到移植肺中。结果成功纯化、扩增MSCs ，传代周期约5 d ，体外传代15代，MSCs仍具有纺锤状形态，保持显著增殖能力及生物学稳定性。流式细胞仪检测结果符合MSCs表型特征。MSCs具有分化为成骨细胞和脂肪细胞的多向分化潜能。可见GFP和DAPI双重标记的MSCs趋化到移植肺中。结论通过全骨髓贴壁培养纯化的MSCs生物学特征稳定；DAPI和GFP双重标记的MSCs能够趋化到移植肺内并存活。
목적관찰체외배양대서골수간충질간세포（MSCs）적생물학특정병연구DAPI화GFP쌍중표기적MSCs향이식폐적추화현상。방법전골수첩벽배양법분리배양대서MSCs ，류식세포법검측세포표형。관찰포괄세포형태학、생장곡선재내적생물학특정。통과정향유도MSCs향성골화성지방분화감정기다향분화잠능。록색형광단백（GFP）화4＇，6-이미기-2-분기신타（DAPI）쌍중표기MSCs ，경미정맥주입수체대서，취폐조직빙동절편，관찰MSCs시부추화도이식폐중。결과성공순화、확증MSCs ，전대주기약5 d ，체외전대15대，MSCs잉구유방추상형태，보지현저증식능력급생물학은정성。류식세포의검측결과부합MSCs표형특정。MSCs구유분화위성골세포화지방세포적다향분화잠능。가견GFP화DAPI쌍중표기적MSCs추화도이식폐중。결론통과전골수첩벽배양순화적MSCs생물학특정은정；DAPI화GFP쌍중표기적MSCs능구추화도이식폐내병존활。
Objective To investigate the biological characteristics of the rat bone marrow mesen-chymal stem cells (MSCs) under culture and study their chemotaxis in vivo after lung transplantation in rats . Methods Rat MSCs were isolated and cultured with direct anchoring method ,and identified by flow cytometry . Their biological characters ,including morphology and cell growth curve were studied . The adipocytic and osteoblastic differentiation abilities were investigated . MSCs were double labeled with 4′,6-diamidino-2-phenylindole (DAPI) and green fluorescent protein (GFP) and injected into rat tail vein after orthotropic lung transplantation . Fluorescence was used to observe their homing to lung trans-plantation recipient by frozen section staining . Results After purification and proliferation ,MSCs were gained and passaged every five days . During 15 generation culture ,MSCs still had spindle-shaped cells and kept significant renewal capacity and biologic stabilities . Flow cytometry test results met the MSC phenotype . MSCs had the abilities to differentiate into adipocyte and osteoblast cells . MSC labeled with double DAPI and GFP can be found in lung transplantation recipient of rats . Conclusion Purified MSCs with direct anchoring method have steady biological characters . MSCs labeled with double DAPI and GFP can survive and be found in lung transplantation recipient of rats .