中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2009年
7期
544-548
,共5页
王敏%裴海云%管利东%南雪%白慈贤%刘慧%李保伟%王韫芳%裴雪涛
王敏%裴海雲%管利東%南雪%白慈賢%劉慧%李保偉%王韞芳%裴雪濤
왕민%배해운%관리동%남설%백자현%류혜%리보위%왕운방%배설도
干细胞%细胞分化%肝细胞%增殖
榦細胞%細胞分化%肝細胞%增殖
간세포%세포분화%간세포%증식
Stem cells%Cell differentiation%Hepatocytes%Proliferation
目的 探讨人脂肪间充质干细胞在改良的诱导体系下向肝细胞的分化和增殖情况,为肝组织工程提供新的种子细胞来源.方法 从人脂肪组织分离出脂肪间充质干细胞,用含有碱性成纤维细胞生长因子、肝细胞生长因子、成纤维细胞生长因子-4的肝细胞诱导液进行诱导,并于诱导7 d后加入抑瘤素M.用细胞计数试剂盒-8法检测整个诱导过程细胞的增殖情况;通过光学显微镜观察诱导细胞的形态变化;用RT-PCR法和免疫荧光法分别检测肝细胞特异性基因和蛋白的表达;并对多种肝细胞特异性功能进行检测.组间比较采用t-test检验.结果 用改良肝细胞诱导液培养的人脂肪间充质干细胞在培养第5、7、14、21天时,细胞数均明显多于用对照培养液培养的细胞(f值分别为6.59、8.69、15.94和24.64,P值均＜0.05).诱导细胞表现出上皮样肝细胞形态,表达肝细胞特异性基因和蛋白;具有多种肝细胞特异性功能,如靛青绿摄取/排泌、糖原合成以及白蛋白分泌功能.结论 人脂肪间充质干细胞在含碱性成纤维细胞生长因子、肝细胞生长因子、成纤维细胞生长因子-4和抑瘤索M的诱导体系中能够分化为更加成熟的具有多种肝细胞特异性功能的细胞,且此诱导体系同时具有促进细胞增殖的作用.
目的 探討人脂肪間充質榦細胞在改良的誘導體繫下嚮肝細胞的分化和增殖情況,為肝組織工程提供新的種子細胞來源.方法 從人脂肪組織分離齣脂肪間充質榦細胞,用含有堿性成纖維細胞生長因子、肝細胞生長因子、成纖維細胞生長因子-4的肝細胞誘導液進行誘導,併于誘導7 d後加入抑瘤素M.用細胞計數試劑盒-8法檢測整箇誘導過程細胞的增殖情況;通過光學顯微鏡觀察誘導細胞的形態變化;用RT-PCR法和免疫熒光法分彆檢測肝細胞特異性基因和蛋白的錶達;併對多種肝細胞特異性功能進行檢測.組間比較採用t-test檢驗.結果 用改良肝細胞誘導液培養的人脂肪間充質榦細胞在培養第5、7、14、21天時,細胞數均明顯多于用對照培養液培養的細胞(f值分彆為6.59、8.69、15.94和24.64,P值均＜0.05).誘導細胞錶現齣上皮樣肝細胞形態,錶達肝細胞特異性基因和蛋白;具有多種肝細胞特異性功能,如靛青綠攝取/排泌、糖原閤成以及白蛋白分泌功能.結論 人脂肪間充質榦細胞在含堿性成纖維細胞生長因子、肝細胞生長因子、成纖維細胞生長因子-4和抑瘤索M的誘導體繫中能夠分化為更加成熟的具有多種肝細胞特異性功能的細胞,且此誘導體繫同時具有促進細胞增殖的作用.
목적 탐토인지방간충질간세포재개량적유도체계하향간세포적분화화증식정황,위간조직공정제공신적충자세포래원.방법 종인지방조직분리출지방간충질간세포,용함유감성성섬유세포생장인자、간세포생장인자、성섬유세포생장인자-4적간세포유도액진행유도,병우유도7 d후가입억류소M.용세포계수시제합-8법검측정개유도과정세포적증식정황;통과광학현미경관찰유도세포적형태변화;용RT-PCR법화면역형광법분별검측간세포특이성기인화단백적표체;병대다충간세포특이성공능진행검측.조간비교채용t-test검험.결과 용개량간세포유도액배양적인지방간충질간세포재배양제5、7、14、21천시,세포수균명현다우용대조배양액배양적세포(f치분별위6.59、8.69、15.94화24.64,P치균＜0.05).유도세포표현출상피양간세포형태,표체간세포특이성기인화단백;구유다충간세포특이성공능,여전청록섭취/배비、당원합성이급백단백분비공능.결론 인지방간충질간세포재함감성성섬유세포생장인자、간세포생장인자、성섬유세포생장인자-4화억류색M적유도체계중능구분화위경가성숙적구유다충간세포특이성공능적세포,차차유도체계동시구유촉진세포증식적작용.
Objective To induce hepatic differentiation of human adipose-derived stem cells (hADSCs) in vitro. Methods hADSCs were isolated from human adipose tissue and treated with improved hepatic medium containing HGF, bFGF and FGF4. After 7 days of culture, OSM was added to the culture media. Cell growth during hepatic differentiation was evaluated by CCK8 assay. Morphology of differentiation was examined under light microscope. Liver speeific genes and proteins were detected by RT-PCR analysis and immunohistochemical staining, respectively. And functional characteristics of hepatocytes were also examined.Results The number ofhADSCs cultured in the improved hepatic media was increased significantly in comparison to hADSCs cultured in control media from 5 days to 21days (t = 6.59, 8.69, 15.94 and 24.64, respectively, P ＜ 0.05). The hADSCs-derived hepatocyte-like cells exhibited hepatocyte morphology, expressed hepatocyte markers, possessed hepatocyte-specific activities, such as uptake and excretion of indocyanine green, glycogen storage and albumin production. Conclusion hADSCs can be induced into hepatocyte-like cells in this differentiation system. And this differentiation system promoted the growth of hADSCs.
