中华骨质疏松和骨矿盐疾病杂志
中華骨質疏鬆和骨礦鹽疾病雜誌
중화골질소송화골광염질병잡지
CHINESE JOURNAL OF OSTEOPOROSIS AND BONE MINERAL RESEARCH
2014年
4期
325-331
,共7页
张红艳%杨乃龙%徐丽丽%陈振
張紅豔%楊迺龍%徐麗麗%陳振
장홍염%양내룡%서려려%진진
人胎盘间充质干细胞%成骨细胞%共培养
人胎盤間充質榦細胞%成骨細胞%共培養
인태반간충질간세포%성골세포%공배양
human placenta mensenchymal stem cells%osteoblasts%co-culture method
目的：观察体外人足月胎盘间充质干细胞（ hPMSCs ）和成骨细胞共培养体系条件下成骨细胞对hPMSCs分化的影响。方法采用胶原酶消化法从人足月胎盘中分离纯化间充质干细胞（ MSCs），检测细胞表面标志物、生长曲线、细胞超微结构及成骨能力并对hPMSCs进行鉴定。共培养组将成骨细胞接种于Tran-swell双层培养皿底层， hPMSCs接种于上层；对照组上层与底层均接种hPMSCs。对诱导后细胞进行碱性磷酸酶染色鉴定。结果胎盘分离细胞经形态、生长速度、细胞表面标志物（ CD44和CD29阳性表达为99％， CD34和CD106为1％），确定为胎盘间充质干细胞；头盖骨分离细胞经碱性磷酸酶染色确定为成骨细胞。采用Transwell共培养hPMSCs 和成骨细胞组碱性磷酸酶活性染色阳性率为（21.7±5.3）％，表现成骨细胞特性，对照组染色呈阴性。结论人足月胎盘含MSCs，与其他来源MSCs生物学特性相似，成骨细胞生长过程提供的微环境对hPMSCs分化为成骨细胞具有诱导促进作用。
目的：觀察體外人足月胎盤間充質榦細胞（ hPMSCs ）和成骨細胞共培養體繫條件下成骨細胞對hPMSCs分化的影響。方法採用膠原酶消化法從人足月胎盤中分離純化間充質榦細胞（ MSCs），檢測細胞錶麵標誌物、生長麯線、細胞超微結構及成骨能力併對hPMSCs進行鑒定。共培養組將成骨細胞接種于Tran-swell雙層培養皿底層， hPMSCs接種于上層；對照組上層與底層均接種hPMSCs。對誘導後細胞進行堿性燐痠酶染色鑒定。結果胎盤分離細胞經形態、生長速度、細胞錶麵標誌物（ CD44和CD29暘性錶達為99％， CD34和CD106為1％），確定為胎盤間充質榦細胞；頭蓋骨分離細胞經堿性燐痠酶染色確定為成骨細胞。採用Transwell共培養hPMSCs 和成骨細胞組堿性燐痠酶活性染色暘性率為（21.7±5.3）％，錶現成骨細胞特性，對照組染色呈陰性。結論人足月胎盤含MSCs，與其他來源MSCs生物學特性相似，成骨細胞生長過程提供的微環境對hPMSCs分化為成骨細胞具有誘導促進作用。
목적：관찰체외인족월태반간충질간세포（ hPMSCs ）화성골세포공배양체계조건하성골세포대hPMSCs분화적영향。방법채용효원매소화법종인족월태반중분리순화간충질간세포（ MSCs），검측세포표면표지물、생장곡선、세포초미결구급성골능력병대hPMSCs진행감정。공배양조장성골세포접충우Tran-swell쌍층배양명저층， hPMSCs접충우상층；대조조상층여저층균접충hPMSCs。대유도후세포진행감성린산매염색감정。결과태반분리세포경형태、생장속도、세포표면표지물（ CD44화CD29양성표체위99％， CD34화CD106위1％），학정위태반간충질간세포；두개골분리세포경감성린산매염색학정위성골세포。채용Transwell공배양hPMSCs 화성골세포조감성린산매활성염색양성솔위（21.7±5.3）％，표현성골세포특성，대조조염색정음성。결론인족월태반함MSCs，여기타래원MSCs생물학특성상사，성골세포생장과정제공적미배경대hPMSCs분화위성골세포구유유도촉진작용。
Objective To establish the co-culture system between human placenta mensenchymal stem cells ( hPMSCs) and osteoblasts in vitro and to study the influence of hPMSCs into osteoblasts in this system .Methods Mes-enchymal stem cells ( MSCs) were isolated and purified from human term placenta using collagenase digestion and identi -fied by cell surface marker , cell growth curve , cell ultrastructure and osteogenesis .Osteoblasts were incubated in Tran-swell double-deck culture dish in the co-culture group .hPMSCs were incubated in the upper layer in high glucose DMEM medium with 10%FBS and identified by alkaline phosphatase staining .hPMSCs were incubated in both layers in the con-trol group.Results Isolated placental cells can be identified as hPMSCs according to morphology , growth rate, cell sur-face markers (CD44 and CD29 99%, CD34 and CD106 1%), ultrastructure, inducing ability , cells isolated from skull can determined as osteoblasts by alkaline phosphatase staining , positive rate of alkaline phosphatase staining in the co-cul-ture group was 21.7%±5.3%, the cells have the characteristics of osteoblasts , but in the control group negative .Con-clusion hPMSCs have the same biological characteristics of MSCs from other sources and microenviroment provided by osteoblasts promotes the differentiation of hPMSCs into osteoblasts .
