感染、炎症、修复
感染、炎癥、脩複
감염、염증、수복
INFECTION， INFLAMMATION， REPAIR
2014年
2期
76-78
,共3页
陈艳%赵洪良%谭志军%赵焕军%张翠萍%付小兵
陳豔%趙洪良%譚誌軍%趙煥軍%張翠萍%付小兵
진염%조홍량%담지군%조환군%장취평%부소병
骨髓间充质干细胞%汗腺%共培养%表型转化
骨髓間充質榦細胞%汗腺%共培養%錶型轉化
골수간충질간세포%한선%공배양%표형전화
Bone marrow mesenchymal stem cell%Sweat gland%Co-culture%Phenotype conversion
目的：观察体外热休克汗腺细胞(SGCs)和人骨髓间充质干细胞(BM-MSCs)共培养体系中 BM-MSCs的形态和表型变化,为进一步表观遗传学表达谱的检测及汗腺诱导关键转录因子的研究提供实验基础。方法：体外分离、培养、扩增人BM-MSCs和 SGCs,成骨和成脂诱导分化以鉴定BM-MSCs 的分化功能。在 Tran-swell间接共培养体系中,培养的BM-MSCs和经47℃高温处理造成热休克的 SGCs 在 Transwell 板中间接共培养；在Transwell+诱导因子共培养体系中,上室的BM-MSCs培养基中添加了汗腺诱导因子(无汗性外胚叶发育不良蛋白、重组人表皮生长因子和胰岛素-转铁蛋白-亚硒酸钠)。监测共培养过程中BM-MSCs的细胞形态变化,免疫荧光法检测诱导后BM-MSCs的表型改变。结果：经与热休克 SGCs 共培养诱导10 d后,部分 BM-MSCs 有由长梭形变为扁平状多边形的趋势,且局部细胞间连接紧密成片。BM-MSCs 诱导前不表达 CEA 和 CK19；BM-MSCs诱导后,Transwell间接共培养体系部分细胞 CEA 和 CK19表达阳性,Transwell+诱导因子共培养体系CEA和CK19阳性细胞数明显多于 Transwell 间接共培养体系。结论：热休克汗腺细胞与 BM-MSCs 在 Tran-swell间接培养以及相关汗腺诱导因子的共培养体系下,BM-MSCs呈现向 SGCs诱导分化趋势。
目的：觀察體外熱休剋汗腺細胞(SGCs)和人骨髓間充質榦細胞(BM-MSCs)共培養體繫中 BM-MSCs的形態和錶型變化,為進一步錶觀遺傳學錶達譜的檢測及汗腺誘導關鍵轉錄因子的研究提供實驗基礎。方法：體外分離、培養、擴增人BM-MSCs和 SGCs,成骨和成脂誘導分化以鑒定BM-MSCs 的分化功能。在 Tran-swell間接共培養體繫中,培養的BM-MSCs和經47℃高溫處理造成熱休剋的 SGCs 在 Transwell 闆中間接共培養；在Transwell+誘導因子共培養體繫中,上室的BM-MSCs培養基中添加瞭汗腺誘導因子(無汗性外胚葉髮育不良蛋白、重組人錶皮生長因子和胰島素-轉鐵蛋白-亞硒痠鈉)。鑑測共培養過程中BM-MSCs的細胞形態變化,免疫熒光法檢測誘導後BM-MSCs的錶型改變。結果：經與熱休剋 SGCs 共培養誘導10 d後,部分 BM-MSCs 有由長梭形變為扁平狀多邊形的趨勢,且跼部細胞間連接緊密成片。BM-MSCs 誘導前不錶達 CEA 和 CK19；BM-MSCs誘導後,Transwell間接共培養體繫部分細胞 CEA 和 CK19錶達暘性,Transwell+誘導因子共培養體繫CEA和CK19暘性細胞數明顯多于 Transwell 間接共培養體繫。結論：熱休剋汗腺細胞與 BM-MSCs 在 Tran-swell間接培養以及相關汗腺誘導因子的共培養體繫下,BM-MSCs呈現嚮 SGCs誘導分化趨勢。
목적：관찰체외열휴극한선세포(SGCs)화인골수간충질간세포(BM-MSCs)공배양체계중 BM-MSCs적형태화표형변화,위진일보표관유전학표체보적검측급한선유도관건전록인자적연구제공실험기출。방법：체외분리、배양、확증인BM-MSCs화 SGCs,성골화성지유도분화이감정BM-MSCs 적분화공능。재 Tran-swell간접공배양체계중,배양적BM-MSCs화경47℃고온처리조성열휴극적 SGCs 재 Transwell 판중간접공배양；재Transwell+유도인자공배양체계중,상실적BM-MSCs배양기중첨가료한선유도인자(무한성외배협발육불량단백、중조인표피생장인자화이도소-전철단백-아서산납)。감측공배양과정중BM-MSCs적세포형태변화,면역형광법검측유도후BM-MSCs적표형개변。결과：경여열휴극 SGCs 공배양유도10 d후,부분 BM-MSCs 유유장사형변위편평상다변형적추세,차국부세포간련접긴밀성편。BM-MSCs 유도전불표체 CEA 화 CK19；BM-MSCs유도후,Transwell간접공배양체계부분세포 CEA 화 CK19표체양성,Transwell+유도인자공배양체계CEA화CK19양성세포수명현다우 Transwell 간접공배양체계。결론：열휴극한선세포여 BM-MSCs 재 Tran-swell간접배양이급상관한선유도인자적공배양체계하,BM-MSCs정현향 SGCs유도분화추세。
Objective:To observe the changes in morphology and phenotype of human bone marrow mesenchymal stem cells (BM-MSCs)after co-cultured with human sweat gland cells (SGCs),in order to provide the experimental basis for the study on sweat gland cells induction-associated key transcription factors. Methods:BM-MSCs and SGCs were isolated,cultured, expanded and identified with the methods of differentiation to adipocytes and osteocytes invitro respectively. In transwell co-cultured experiment,BM-MSCs were induced into SGCs by indirect co-culture with 47 ℃heat-shocked confluent SGCs using transwell plate.But in transwell+inducing factors co-culture experiment,BM-MSCs culture medium was supplemented with series of inducing factors,including EDA-A1 ,recombinant human epidermal growth factor and ITS. After being co-cultured, morphological and phenotypic changes in BM-MSCs were observed. The antigen expressions of BM-MSCs were detected by immunofluorescence. Results:After co-cultured with heat-shock SGCs for 10 days,the shape of a part of MSCs changed from long-clostridial shape to flat and polygonal. Non-induced BM-MSCs were negative for CEA and CK1 9. Induced BM-MSCs were positive for CEA and CK19,and the positive cells of transwell + inducing factors co-culture was more abundant than that of transwell co-culture. Conclusions:MSCs could differentiate into sweat gland-like cells at phenotype and shape by a proper in vitro co-culture system of BM-MSCs and SGCs with or without inducing factors,and it was more efficient when they were co-cultured with inducing factors.
