国际生物医学工程杂志
國際生物醫學工程雜誌
국제생물의학공정잡지
INTERNATIONAL JOURNAL OF BIOMEDICAL ENGINEERING
2010年
3期
163-166,封3
,共5页
王宁%邢万红%李新华%陈平
王寧%邢萬紅%李新華%陳平
왕저%형만홍%리신화%진평
骨髓间充质干细胞%心肌细胞%组织工程
骨髓間充質榦細胞%心肌細胞%組織工程
골수간충질간세포%심기세포%조직공정
Bone marrow mesenchymal stem cells%Cardiomyocyte%Tissue engineering
目的 探讨大鼠骨髓间充质干细胞(BMSCs)向心肌定向诱导分化及诱导后BMSCs体外构建工程化心肌组织的可行性.方法 用含10μmol/L 5-氮胞苷(5-Aza)的完全培养液(LG-DMEM,10%胎牛血清,100U/mL青霉素,100μg/mL链霉素,10μg/L碱性成纤维细胞生长因子)孵育第3代BMSsCs 24h后改用完全培养基培养4周,采用未处理组作为阴性对照.采用免疫细胞化学方法检测心肌肌钙蛋白T(cTnT)、α-肌动蛋白(α-Actin)的表达;逆转录-聚合酶链反应(RT-PCR)检测早期心肌形成转录因子GATA-4、Nkx2.5、TDGF-1基因的表达.将已诱导4周的BMSCs接种于多聚赖氨酸包被的脱细胞牛心包生物支架上培养2周后检测.结果 诱导组细胞向心肌细胞转化,表达cTnT、α-Actin,对照组不表达cTnT、α-Actin;RT-PCR检测BMSCs诱导后表达早期心肌形成转录因子GATA-4、Nkx2.5、TDGF-1等基因;诱导后的BMSCs黏附于脱细胞牛心包生物支架表面,生长良好.结论 大鼠BMSCs可向心肌细胞定向诱导分化,与脱细胞牛心包生物支架黏附良好,有望构建理想的组织工程化心肌.
目的 探討大鼠骨髓間充質榦細胞(BMSCs)嚮心肌定嚮誘導分化及誘導後BMSCs體外構建工程化心肌組織的可行性.方法 用含10μmol/L 5-氮胞苷(5-Aza)的完全培養液(LG-DMEM,10%胎牛血清,100U/mL青黴素,100μg/mL鏈黴素,10μg/L堿性成纖維細胞生長因子)孵育第3代BMSsCs 24h後改用完全培養基培養4週,採用未處理組作為陰性對照.採用免疫細胞化學方法檢測心肌肌鈣蛋白T(cTnT)、α-肌動蛋白(α-Actin)的錶達;逆轉錄-聚閤酶鏈反應(RT-PCR)檢測早期心肌形成轉錄因子GATA-4、Nkx2.5、TDGF-1基因的錶達.將已誘導4週的BMSCs接種于多聚賴氨痠包被的脫細胞牛心包生物支架上培養2週後檢測.結果 誘導組細胞嚮心肌細胞轉化,錶達cTnT、α-Actin,對照組不錶達cTnT、α-Actin;RT-PCR檢測BMSCs誘導後錶達早期心肌形成轉錄因子GATA-4、Nkx2.5、TDGF-1等基因;誘導後的BMSCs黏附于脫細胞牛心包生物支架錶麵,生長良好.結論 大鼠BMSCs可嚮心肌細胞定嚮誘導分化,與脫細胞牛心包生物支架黏附良好,有望構建理想的組織工程化心肌.
목적 탐토대서골수간충질간세포(BMSCs)향심기정향유도분화급유도후BMSCs체외구건공정화심기조직적가행성.방법 용함10μmol/L 5-담포감(5-Aza)적완전배양액(LG-DMEM,10%태우혈청,100U/mL청매소,100μg/mL련매소,10μg/L감성성섬유세포생장인자)부육제3대BMSsCs 24h후개용완전배양기배양4주,채용미처리조작위음성대조.채용면역세포화학방법검측심기기개단백T(cTnT)、α-기동단백(α-Actin)적표체;역전록-취합매련반응(RT-PCR)검측조기심기형성전록인자GATA-4、Nkx2.5、TDGF-1기인적표체.장이유도4주적BMSCs접충우다취뢰안산포피적탈세포우심포생물지가상배양2주후검측.결과 유도조세포향심기세포전화,표체cTnT、α-Actin,대조조불표체cTnT、α-Actin;RT-PCR검측BMSCs유도후표체조기심기형성전록인자GATA-4、Nkx2.5、TDGF-1등기인;유도후적BMSCs점부우탈세포우심포생물지가표면,생장량호.결론 대서BMSCs가향심기세포정향유도분화,여탈세포우심포생물지가점부량호,유망구건이상적조직공정화심기.
Objective To investigate the feasibility of inducing rat bone marrow mesenchymal stem cells (BMSCs)into myocardial cells and the approach of applying the induced MSCs to construct myocardial tissue in vitro. Methods The third generation of BMSCs were incubated in complete culture medium (LG-DMEM,10%FBS,100U/mL penicillin, 100μg/mL streptomycin, 10μg/L bFGF), in which 10μmol/L 5-Aza was added, for 24 hours. Then the BMSCs were switched to complete medium and cultured for 4 weeks as the experimental group. The other group without incubation was cultured in complete medium as the control. cTnT and a-Actin were tested by immunocytochemistry method and the early myocardial transcription factor genes GATA-4、Nkx2.5、TDGF-1 were tested by reverse transcription polymerase chain reaction(PT-PCR). The induced BMSCs were planted onto the acellular bovine pericardium bio-stent which were coated by polylysine(PLYS) and cultured for 2 weeks before being tested. Results The experimental group showed morphological change to car-diomyocytes and expressed cTnT and a-Actin, while the control group did not express cTnT and a-Actin. The experimental group expressed the early myocardial transcription factor GATA-4, Nkx2.5, TDGF-1 gene. The induced BMSCs adhered to acellular bovine pericardium bio-stent surface and grew well. Conclusion BMSCs can be induced into cardiomyocyte in vitro and they adhere well with acellular bovine pericardium bio-stent. BMSCs have the potential to form an ideal cardiac muscle tissue.
