重庆医学
重慶醫學
중경의학
CHONGQING MEDICAL JOURNAL
2014年
2期
147-149,151
,共4页
成纤维细胞%真皮%波形蛋白%角蛋白19%诱导分化
成纖維細胞%真皮%波形蛋白%角蛋白19%誘導分化
성섬유세포%진피%파형단백%각단백19%유도분화
fibroblast%dermis%Vimentin%keratin-19%induction differentiation
目的：体外分离培养人包皮成纤维细胞（HFF），探讨其成骨、成脂、成软骨分化潜能，为骨创伤或其他修复治疗提供新的种子细胞来源。方法采用机械法结合酶消化法分离、培养 HFF。取第3代细胞用免疫细胞化学法检测其波形蛋白（Vim-entin）和角蛋白19（CK19）的表达。通过向成骨、成脂、成软骨细胞诱导分化（诱导培养基根据说明书配制），鉴定其多向分化潜能。结果原代细胞24 h内完全贴壁，细胞形态不规则，多为圆形或多角形，传代后细胞形态均一，基本为长梭形，鱼群样、漩涡样生长。免疫细胞化学结果显示该细胞表达Vimentin ，不表达CK19，经诱导可以向成骨、成脂、软骨细胞方向分化。结论 HFF在特定条件能多向分化，其本身也参与骨折愈合的全过程，加之来源方便，增殖能力强，有望为骨折或较大骨缺损患者修复治疗及组织工程学提供理想种子细胞。
目的：體外分離培養人包皮成纖維細胞（HFF），探討其成骨、成脂、成軟骨分化潛能，為骨創傷或其他脩複治療提供新的種子細胞來源。方法採用機械法結閤酶消化法分離、培養 HFF。取第3代細胞用免疫細胞化學法檢測其波形蛋白（Vim-entin）和角蛋白19（CK19）的錶達。通過嚮成骨、成脂、成軟骨細胞誘導分化（誘導培養基根據說明書配製），鑒定其多嚮分化潛能。結果原代細胞24 h內完全貼壁，細胞形態不規則，多為圓形或多角形，傳代後細胞形態均一，基本為長梭形，魚群樣、漩渦樣生長。免疫細胞化學結果顯示該細胞錶達Vimentin ，不錶達CK19，經誘導可以嚮成骨、成脂、軟骨細胞方嚮分化。結論 HFF在特定條件能多嚮分化，其本身也參與骨摺愈閤的全過程，加之來源方便，增殖能力彊，有望為骨摺或較大骨缺損患者脩複治療及組織工程學提供理想種子細胞。
목적：체외분리배양인포피성섬유세포（HFF），탐토기성골、성지、성연골분화잠능，위골창상혹기타수복치료제공신적충자세포래원。방법채용궤계법결합매소화법분리、배양 HFF。취제3대세포용면역세포화학법검측기파형단백（Vim-entin）화각단백19（CK19）적표체。통과향성골、성지、성연골세포유도분화（유도배양기근거설명서배제），감정기다향분화잠능。결과원대세포24 h내완전첩벽，세포형태불규칙，다위원형혹다각형，전대후세포형태균일，기본위장사형，어군양、선와양생장。면역세포화학결과현시해세포표체Vimentin ，불표체CK19，경유도가이향성골、성지、연골세포방향분화。결론 HFF재특정조건능다향분화，기본신야삼여골절유합적전과정，가지래원방편，증식능력강，유망위골절혹교대골결손환자수복치료급조직공정학제공이상충자세포。
Objective To separate and culture human foreskin fibroblasts (HFF) in vitro and to investigate the differentiation potentials of osteogenesis ,adipogenesis and chondrogenesis to provide the new source of seed cells for bone trauma or other repair treatment .Methods The methanical method combined with the enzyme digestion method was adopted to separate and culture HFF .The third generation cells were taken for detecting the expression of Vimentin and CK 19 by the immunocytochemical method . By the induction differentiation to osteoblast ,adipoblast and chondroblast(the induction medium was prepared according to the in-structions) ,the multi-directional differentiation potentials were identified .Results The primary cells were completely adherent within 24 h ,the cell shapes were irregular ,most of cells were round or polygonal ,after passage ,the cellular shapes were uniform ,u-sually long spindle-shape ,grew in fish shoal-like and swirl-like .The immunocytochemical results showed that these cells expressed Vimentin and not expressed CK19 .After being cultured in inducing differentiation media ,the cells were differentiated into the direc-tions of osteoblasts ,adipoblasts and chondroblasts .Conclusion HFF can multi-directionally differentiate under certain condition , which itself also participates in the whole process of fracture healing ,together with a convenient source and high proliferation abili-ty ,HFF is expected to provide the ideal seed cells for the repair treatment of fracture or larger bone defect and the tissue engineer-ing .
