中华整形外科杂志
中華整形外科雜誌
중화정형외과잡지
CHINESE JOURNAL OF PLASTIC SURGERY
2013年
4期
268-272
,共5页
杨卫玺%欧阳溪%宋云庆%张晓光%张军
楊衛璽%歐暘溪%宋雲慶%張曉光%張軍
양위새%구양계%송운경%장효광%장군
结缔组织生长因子%间充质干细胞%成纤维细胞
結締組織生長因子%間充質榦細胞%成纖維細胞
결체조직생장인자%간충질간세포%성섬유세포
Connective tissue growth factor%Mesenchymal stem cells%Fibroblast
目的 探究胎盘间充质干细胞(mesenchymal stem cells,MSCs)向皮肤成纤维细胞分化的潜能,以及其用于皮肤创伤修复及作为种子细胞的可能性.方法 酶消化法分离得到胎盘MSCs,体外培养扩增,用流式细胞术和成骨、成脂诱导进行MSCs鉴定.加入含50 μg/ml维生素C和100 ng/ml结缔组织生长因子(connective tissue growth factor,CTGF)的DMEM/F12诱导胎盘MSCs 16 d,拍照记录形态变化,免疫荧光方法检测诱导前、后细胞内波形蛋白、人纤维母细胞表面抗原(human fibroblast surface protein,FSP)、Ⅰ型和Ⅲ型胶原、结蛋白及层粘连蛋白的表达变化,同时检测诱导后成骨、成脂分化能力变化.冻存复苏诱导后细胞,台盼蓝法检测细胞活率.结果 胎盘MSCs经CTGF诱导之后,具有明显的成纤维细胞形态,波形蛋白、FSP-1、Ⅰ型和Ⅲ型胶原及层粘连蛋白表达升高,且表达皮肤成纤维细胞特异性蛋白结蛋白,成骨、成脂分化能力减弱,胎盘MSCs成功诱导为皮肤成纤维细胞.诱导细胞冻存复苏后细胞活率大于90％.结论 胎盘MSCs在体外可经CTGF诱导为皮肤成纤维细胞,胎盘MSCs有潜在的用于皮肤创伤修复价值,并可作为皮肤成纤维细胞的种子细胞保存.
目的 探究胎盤間充質榦細胞(mesenchymal stem cells,MSCs)嚮皮膚成纖維細胞分化的潛能,以及其用于皮膚創傷脩複及作為種子細胞的可能性.方法 酶消化法分離得到胎盤MSCs,體外培養擴增,用流式細胞術和成骨、成脂誘導進行MSCs鑒定.加入含50 μg/ml維生素C和100 ng/ml結締組織生長因子(connective tissue growth factor,CTGF)的DMEM/F12誘導胎盤MSCs 16 d,拍照記錄形態變化,免疫熒光方法檢測誘導前、後細胞內波形蛋白、人纖維母細胞錶麵抗原(human fibroblast surface protein,FSP)、Ⅰ型和Ⅲ型膠原、結蛋白及層粘連蛋白的錶達變化,同時檢測誘導後成骨、成脂分化能力變化.凍存複囌誘導後細胞,檯盼藍法檢測細胞活率.結果 胎盤MSCs經CTGF誘導之後,具有明顯的成纖維細胞形態,波形蛋白、FSP-1、Ⅰ型和Ⅲ型膠原及層粘連蛋白錶達升高,且錶達皮膚成纖維細胞特異性蛋白結蛋白,成骨、成脂分化能力減弱,胎盤MSCs成功誘導為皮膚成纖維細胞.誘導細胞凍存複囌後細胞活率大于90％.結論 胎盤MSCs在體外可經CTGF誘導為皮膚成纖維細胞,胎盤MSCs有潛在的用于皮膚創傷脩複價值,併可作為皮膚成纖維細胞的種子細胞保存.
목적 탐구태반간충질간세포(mesenchymal stem cells,MSCs)향피부성섬유세포분화적잠능,이급기용우피부창상수복급작위충자세포적가능성.방법 매소화법분리득도태반MSCs,체외배양확증,용류식세포술화성골、성지유도진행MSCs감정.가입함50 μg/ml유생소C화100 ng/ml결체조직생장인자(connective tissue growth factor,CTGF)적DMEM/F12유도태반MSCs 16 d,박조기록형태변화,면역형광방법검측유도전、후세포내파형단백、인섬유모세포표면항원(human fibroblast surface protein,FSP)、Ⅰ형화Ⅲ형효원、결단백급층점련단백적표체변화,동시검측유도후성골、성지분화능력변화.동존복소유도후세포,태반람법검측세포활솔.결과 태반MSCs경CTGF유도지후,구유명현적성섬유세포형태,파형단백、FSP-1、Ⅰ형화Ⅲ형효원급층점련단백표체승고,차표체피부성섬유세포특이성단백결단백,성골、성지분화능력감약,태반MSCs성공유도위피부성섬유세포.유도세포동존복소후세포활솔대우90％.결론 태반MSCs재체외가경CTGF유도위피부성섬유세포,태반MSCs유잠재적용우피부창상수복개치,병가작위피부성섬유세포적충자세포보존.
Objective To investigate the possibility of placenta mesenchymal stem cells (PMSCs) differentiation into dermal fibroblast,and the potency of PMSCs used in cutaneous wound healing and stored as seed cells.Methods Enzyme digestion method was used to obtain PMSCs,and PMSCs were amplified after culture in vitro.Flow cytometry assay,osteogenic and adipogenic differentiation were done for MSCs identification.The induction medium composed of DMEM/F12 + 50 μg/ml VC + 100 ng/ml connective tissue growth factor (CTGF) was added into the 24-well plate for 16 days induction period.Pictures were taken to record morphologic change.Immunofluorescence tests were performed to detect Vimentin,FSP-1,collagen Ⅰ,collagen Ⅲ,desmin and laminin expression before and after induction.At the same time osteogenic and adipogenic differentiation were used to assay the differentiation ability change after induction.The induced dermal fibroblasts were frozen in liquid nitrogenand and recovery and trypan blue was used to detect cell viability.Results After CTGF induction,PMSCs got obvious fibroblasts morphology,the protein level of Vimentin,FSP-1,collagen Ⅰ,collagen Ⅲ and Laminin increased,PMSCs started to express Desmin,the dermal fibroblasts specific proteins,and osteogenic and adipogenic differentiation ability was diminished.PMSCs were successfully induced into dermal fibroblasts,and these induced cells could get a high cell viability (more than 90％) after recovery.Conclusions PMSCs could be induced into dermal fibroblasts by CTGF in vitro.PMSCs have the potential application in skin wound healing,and can be used as seed cells of dermal fibroblasts.
