中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
20期
3133-3138
,共6页
田茂%朴海南%陈宇%赵琦%潘心
田茂%樸海南%陳宇%趙琦%潘心
전무%박해남%진우%조기%반심
组织构建%组织工程%干细胞%骨髓间充质干细胞%聚乙交酯-聚己内酯共聚物%补片%心肌梗死
組織構建%組織工程%榦細胞%骨髓間充質榦細胞%聚乙交酯-聚己內酯共聚物%補片%心肌梗死
조직구건%조직공정%간세포%골수간충질간세포%취을교지-취기내지공취물%보편%심기경사
stem cells%mesenchymal stem cells%myocardial infarction%stents%staining and labeling
背景:到目前为止,工程化心肌组织仍然面临很多的问题。研究证实,骨髓间充质干细胞可分化为心肌样细胞;聚乙交酯、聚己内酯为常用的人工高分子材料,生物相容性良好。<br> 目的:观察体外构建骨髓间充质干细胞-聚乙交酯-聚己内酯共聚物补片在正常心肌组织和梗死后心肌组织中的生长情况。<br> 方法:SD大鼠骨髓间充质干细胞采用贴壁分离筛选法进行分离、培养,选取第3代进行体外DAPI标记。制作骨髓间充质干细胞悬液(2×106/cm2)种植于聚乙交酯-聚己内酯共聚物上形成骨髓间充质干细胞-聚乙交酯-聚己内酯共聚物补片。培养48 h在电镜下观察;苏木精-伊红染色后在光学显微镜下观察。大鼠结扎左冠状动脉前降支制作心肌梗死模型。将补片植入到正常心肌组织和梗死后心肌组织培养5周,行病理学检查了解骨髓间充质干细胞在组织中的存活情况。<br> 结果与结论:光学显微镜和电镜观察结果示,骨髓间充质干细胞在聚乙交酯-聚己内酯共聚物支架中呈三维生长,细胞与支架黏附良好。在激光共聚焦显微镜下,对比观察补片植入正常心肌组织后第1周和第5周切片结果,同第1周相比,第5周的心肌组织内出现DAPI标记的骨髓间充质干细胞;心肌梗死区出现DAPI标记的骨髓间充质干细胞。苏木精-伊红染色结果示,梗死区出现骨髓间充质干细胞。提示骨髓间充质干细胞在聚乙交酯-聚己内酯共聚物支架上贴附生长,黏附良好。骨髓间充质干细胞-聚乙交酯-聚己内酯共聚物补片可用于心肌组织修复。
揹景:到目前為止,工程化心肌組織仍然麵臨很多的問題。研究證實,骨髓間充質榦細胞可分化為心肌樣細胞;聚乙交酯、聚己內酯為常用的人工高分子材料,生物相容性良好。<br> 目的:觀察體外構建骨髓間充質榦細胞-聚乙交酯-聚己內酯共聚物補片在正常心肌組織和梗死後心肌組織中的生長情況。<br> 方法:SD大鼠骨髓間充質榦細胞採用貼壁分離篩選法進行分離、培養,選取第3代進行體外DAPI標記。製作骨髓間充質榦細胞懸液(2×106/cm2)種植于聚乙交酯-聚己內酯共聚物上形成骨髓間充質榦細胞-聚乙交酯-聚己內酯共聚物補片。培養48 h在電鏡下觀察;囌木精-伊紅染色後在光學顯微鏡下觀察。大鼠結扎左冠狀動脈前降支製作心肌梗死模型。將補片植入到正常心肌組織和梗死後心肌組織培養5週,行病理學檢查瞭解骨髓間充質榦細胞在組織中的存活情況。<br> 結果與結論:光學顯微鏡和電鏡觀察結果示,骨髓間充質榦細胞在聚乙交酯-聚己內酯共聚物支架中呈三維生長,細胞與支架黏附良好。在激光共聚焦顯微鏡下,對比觀察補片植入正常心肌組織後第1週和第5週切片結果,同第1週相比,第5週的心肌組織內齣現DAPI標記的骨髓間充質榦細胞;心肌梗死區齣現DAPI標記的骨髓間充質榦細胞。囌木精-伊紅染色結果示,梗死區齣現骨髓間充質榦細胞。提示骨髓間充質榦細胞在聚乙交酯-聚己內酯共聚物支架上貼附生長,黏附良好。骨髓間充質榦細胞-聚乙交酯-聚己內酯共聚物補片可用于心肌組織脩複。
배경:도목전위지,공정화심기조직잉연면림흔다적문제。연구증실,골수간충질간세포가분화위심기양세포;취을교지、취기내지위상용적인공고분자재료,생물상용성량호。<br> 목적:관찰체외구건골수간충질간세포-취을교지-취기내지공취물보편재정상심기조직화경사후심기조직중적생장정황。<br> 방법:SD대서골수간충질간세포채용첩벽분리사선법진행분리、배양,선취제3대진행체외DAPI표기。제작골수간충질간세포현액(2×106/cm2)충식우취을교지-취기내지공취물상형성골수간충질간세포-취을교지-취기내지공취물보편。배양48 h재전경하관찰;소목정-이홍염색후재광학현미경하관찰。대서결찰좌관상동맥전강지제작심기경사모형。장보편식입도정상심기조직화경사후심기조직배양5주,행병이학검사료해골수간충질간세포재조직중적존활정황。<br> 결과여결론:광학현미경화전경관찰결과시,골수간충질간세포재취을교지-취기내지공취물지가중정삼유생장,세포여지가점부량호。재격광공취초현미경하,대비관찰보편식입정상심기조직후제1주화제5주절편결과,동제1주상비,제5주적심기조직내출현DAPI표기적골수간충질간세포;심기경사구출현DAPI표기적골수간충질간세포。소목정-이홍염색결과시,경사구출현골수간충질간세포。제시골수간충질간세포재취을교지-취기내지공취물지가상첩부생장,점부량호。골수간충질간세포-취을교지-취기내지공취물보편가용우심기조직수복。
BACKGROUND:So far, engineered myocardium is stil facing many problems. Research has demonstrated that bone marrow mesenchymal stem cells can be differentiated into myocardial cells. Polyglycolide and polycaprolactone are commonly used artificial polymers, which have good biocompatibility. <br> OBJECTIVE:To observe the growth of the poly(glycolic acid)/poly(e-caprolactone) copolymer patch in vitro in normal myocardium and infarcted myocardium. <br> METHODS:Bone marrow mesenchymal stem cells in Sprague-Dawley rats were separated using adherent separation and selection method, cultured in vitro. The third passage was labeled with DAPI. The bone marrow mesenchymal stem cellsuspensions (2×106/cm2) were produced and planted on poly(glycolic acid)/poly(e-caprolactone) copolymer scaffolds to form poly(glycolic acid)/poly(e-caprolactone) copolymer patch. After culturing for 48 hours, the specimens were observed under electron microscope, stained with hematoxylin and eosin, and then observed under light microscope. Rat models of myocardial infarction were established by ligating left anterior descending coronary artery. Poly(glycolic acid)/poly(e-caprolactone) copolymer patch was implanted into the normal and infarcted myocardium for 5 weeks. The survival of bone marrow mesenchymal stem cells was determined by the detection of pathology. <br> RESULTS AND CONCLUSION:Results of light microscope and electron microscope demonstrated that bone marrow mesenchymal stem cells grew three-dimensional y on poly(glycolic acid)/poly(e-caprolactone) copolymer patch. cells and patch were adhesive wel . Under laser confocal microscopy, compared with the first week, bone marrow mesenchymal stem cells were marked by DAPI in the myocardium at the fifth week. There were bone marrow mesenchymal stem cells marked by DAPI in the infracted area. Results of hematoxylin-eosin staining exhibited that bone marrow mesenchymal stem cells were detected in the infarct area. These results suggested that bone marrow mesenchymal stem cells adhered to the poly(glycolic acid)/poly(e-caprolactone) copolymer stent wel . The complexes of poly(glycolic acid)/poly(e-caprolactone) copolymer and bone marrow mesenchymal stem cells can be used for reparation of myocardium.