CAJ | 학술논문

目的分离、培养人脂肪间充质干细胞(hADSCs),研究其在体内外向类肝样细胞转化的能力.方法采用Ⅰ型胶原酶消化皮下脂肪,贴壁分选去除造血细胞的方法获得较纯的干细胞.通过对形态观察、表面分子鉴定、透射电镜内部结构扫描以及成脂、成骨定向诱导对脂肪间充质干细胞进行鉴定.用EGF、FGF、OSM、HGF、TSA细胞因子体外诱导hADSCs向肝样细胞分化.尾静脉输注hADSCs至急性肝损伤裸鼠,观察hADSCs在裸鼠肝脏内定植、向肝样细胞分化情况,以及hADSCs对裸鼠肝损伤的保护作用.结果分离获得的细胞呈长梭形贴壁生长,低表达CD34、CD45,高表达CD73、CD90、CD105,具有多向分化潜能.诱导细胞表现出肝样细胞形态,表达AFP、alb,并且和肝细胞一样具有吸脂性.体内诱导:移植细胞在裸鼠体内生存、分布,并且细胞表达人alb,细胞体积较裸鼠肝脏细胞大,形态不均一,与人肝脏细胞形态有差异.同时研究发现hADSCs可改善急性肝损伤裸鼠肝功能,降低转氨酶.结论人脂肪间充质干细胞可在体内外向肝样细胞分化,分化的细胞具有肝细胞样生物学性状.hADSCs对裸鼠急性肝损伤具有一定的治疗作用.
목적 분리、배양인지방간충질간세포(hADSCs),연구기재체내외향류간양세포전화적능력.방법 채용Ⅰ형효원매소화피하지방,첩벽분선거제조혈세포적방법획득교순적간세포.통과대형태관찰、표면분자감정、투사전경내부결구소묘이급성지、성골정향유도대지방간충질간세포진행감정.용EGF、FGF、OSM、HGF、TSA세포인자체외유도hADSCs향간양세포분화.미정맥수주hADSCs지급성간손상라서,관찰hADSCs재라서간장내정식、향간양세포분화정황,이급hADSCs대라서간손상적보호작용.결과 분리획득적세포정장사형첩벽생장,저표체CD34、CD45,고표체CD73、CD90、CD105,구유다향분화잠능.유도세포표현출간양세포형태,표체AFP、alb,병차화간세포일양구유흡지성.체내유도:이식세포재라서체내생존、분포,병차세포표체인alb,세포체적교라서간장세포대,형태불균일,여인간장세포형태유차이.동시연구발현hADSCs가개선급성간손상라서간공능,강저전안매.결론 인지방간충질간세포가재체내외향간양세포분화,분화적세포구유간세포양생물학성상.hADSCs대라서급성간손상구유일정적치료작용.
Objective To isolate and culture human adipose derived mesenchynal stem cells (hADSCs) and study the potential of hADSCs to differentiate into hepatocyte-like cells. Methods To ensure the removal of contaminating hematopoietic cells, hADSCs were selected based on plastic adherence. Cell surface antigen was confirmed by flow cytometry; ultramicrostructure was detected by transmission electron microscopy; adipogenic and osteogenic differentiation of hADSCs was analyzed by oil Red O staining and yon Kossa staining. hADSCs were exposed to differentiation medium containing EGF,FGF,OSM, HGF,TSA in vitro. 10% CCl4 (100 μ1/20 g body weight )was injected into immune-deficient BALB/c-nu mice and hADSCs (5 × 105 cells) were simultaneously administrated from the tail vein. Blood samples were collected and concentration of aminotransferase and direct bilirubin were detected. Administration without hADSCs was used as a control. One month later, we sacrificed the mice and liver sections were examined by histochemical immunofluorescence with human ALB specific antibodies. Results hADSCs exhibited fibroblast-like morphology, and expressed CD73, CD90,CD105, and were lacking of CD34 and CD45. Adipogenic and osteogenic differentiation showed that hADSCs have the capacity of multidifferentiation. The differentiated cells showed hepatocyte-like cell morphologies and hepatocyte-specific markers including albumin (alb) and α-fetoprotein (AFP). The bioactivity assays revealed that these hepatocyte-like cells could uptake low-density lipoprotein (LDL).Histochemical immunofluorescence showed that hADSCs were incorporated into injured livers. Some human alb-positive cells were found in liver sections after implantation of undifferentiated hADSCs. Transaminase activity in the experimental group was lower than in the control group. Conclution hADSCs can differentiated into functional hepatocyte-like cells and can relieve CCl4 induced BALB/c-nu acute liver injury.