The translation of current tissue engineering approaches to clinical application is somehow limited by the use of scaffolding materials.Recently a number of in vitro scaffold-free three-dimensional culture techniques ...The translation of current tissue engineering approaches to clinical application is somehow limited by the use of scaffolding materials.Recently a number of in vitro scaffold-free three-dimensional culture techniques have been developed.These techniques realize the assembly of tissue-like structures including but not limited to spheroids,blood vessels and cartilage.In particular,cells can now self-assemble to form planar tissue-like structures at the interface of an aqueous-two-phase system(ATPS).The unique advantage of this technique is that without a solid substrate,planar tissue-like structures can now be assembled rapidly with very simple procedures.This technique can potentially be very useful for tissue engineering in eye because of its ability to direct cells to form monolayer.In this talk,we will introduce what ATPS is and its current applications in biomedical research.We will then present an approach to assemble cell sheets in ATPS using both primary cells isolated from porcine eyes and other cell lines.The physiological relevance of these eye-related cell sheets as well as their potentials in ophthalmic research and applications will be discussed.展开更多
细胞片技术是应用组织工程方法使培养细胞从培养表面分离而形成含有细胞外基质的一层完整片状结构,弥补了传统组织工程技术的不足,是获取种子细胞以及对种子细胞进行转移的一项新技术。为探讨体外生长分化因子-5(GDF5)基因转染修饰的BM...细胞片技术是应用组织工程方法使培养细胞从培养表面分离而形成含有细胞外基质的一层完整片状结构,弥补了传统组织工程技术的不足,是获取种子细胞以及对种子细胞进行转移的一项新技术。为探讨体外生长分化因子-5(GDF5)基因转染修饰的BMSCs细胞片与GDF5转基因BMSCs负载的PLGA支架形成的共聚物修复兔甲状软骨缺损的效果,实验通过腺病毒转染GDF5基因至四代兔BMSCs,温度敏感性培养皿制备GDF5转基因细胞片并与负载有转染GDF5基因BMSCs的PLGA支架复合,移植至同种兔甲状软骨缺损处,分别于术后4、8周行大体观察和组织学检测其修复效果。实验分3组:(A)转基因细胞片包裹负载有转基因BMSCs的PLGA支架组;(B)负载有转基因BMSCs的PLGA支架组;(C)负载BMSCs的PLGA支架组。结果显示,体外成功收获了完整的GDF5转基因细胞片,Real time PCR检测到GDF5 mRNA的表达,行大体组织的II型胶原免疫组化和阿利新蓝染色显示:A组和B组均表达II型胶原和糖胺聚糖(GAG),但A组表达高于B组,有统计学意义(P<0.05)。由此可得,转基因细胞片包裹负载转基因BMSCs PLGA支架较传统转基因BMSCs负载PLGA支架方法具有更加优越的成软骨能力,能更有效地促进软骨缺损的修复。展开更多
目的研究利用骨髓基质细胞膜片复合聚乳乙醇酸（ploy of lactic-co-glycolic acid，PLGA）支撑体，在生物反应器条件下体外构建管状软骨的可行性。方法分离兔骨髓基质细胞，高密度连续培养，转化生长因子-1诱导构建成干细胞膜片，制作...目的研究利用骨髓基质细胞膜片复合聚乳乙醇酸（ploy of lactic-co-glycolic acid，PLGA）支撑体，在生物反应器条件下体外构建管状软骨的可行性。方法分离兔骨髓基质细胞，高密度连续培养，转化生长因子-1诱导构建成干细胞膜片，制作圆柱状PLGA支撑体，将细胞膜片均匀缠绕在表面。静置孵育14d，使细胞膜片与PLGA相互贴附后，进入生物反应器动态培养8周后，取出标本。从大体形态、组织学结构、蛋白多糖含量以及生物力学性能等方面评价形成软骨的理化特性。结果通过此策略构建的软骨外观与天然软骨组织非常相似，保持着良好的管状外形，颜色呈乳白色，有光泽，质地均匀，弹性好，具有中等偏软的硬度。组织学结果显示总体结构呈现软骨样结构，HE染色可见软骨样细胞分布于细胞陷窝之中，周围是均匀的细胞外基质，番红-O染色可见细胞外基质着色为鲜红色，提示蛋白多糖含量丰富，有大量软骨基质物产生。结论细胞膜片复合支撑体策略能形成管状形态的软骨组织，为气管软骨的再造提供了新的方法，有可能解决气管缺损的临床难题。展开更多
文摘The translation of current tissue engineering approaches to clinical application is somehow limited by the use of scaffolding materials.Recently a number of in vitro scaffold-free three-dimensional culture techniques have been developed.These techniques realize the assembly of tissue-like structures including but not limited to spheroids,blood vessels and cartilage.In particular,cells can now self-assemble to form planar tissue-like structures at the interface of an aqueous-two-phase system(ATPS).The unique advantage of this technique is that without a solid substrate,planar tissue-like structures can now be assembled rapidly with very simple procedures.This technique can potentially be very useful for tissue engineering in eye because of its ability to direct cells to form monolayer.In this talk,we will introduce what ATPS is and its current applications in biomedical research.We will then present an approach to assemble cell sheets in ATPS using both primary cells isolated from porcine eyes and other cell lines.The physiological relevance of these eye-related cell sheets as well as their potentials in ophthalmic research and applications will be discussed.
文摘细胞片技术是应用组织工程方法使培养细胞从培养表面分离而形成含有细胞外基质的一层完整片状结构,弥补了传统组织工程技术的不足,是获取种子细胞以及对种子细胞进行转移的一项新技术。为探讨体外生长分化因子-5(GDF5)基因转染修饰的BMSCs细胞片与GDF5转基因BMSCs负载的PLGA支架形成的共聚物修复兔甲状软骨缺损的效果,实验通过腺病毒转染GDF5基因至四代兔BMSCs,温度敏感性培养皿制备GDF5转基因细胞片并与负载有转染GDF5基因BMSCs的PLGA支架复合,移植至同种兔甲状软骨缺损处,分别于术后4、8周行大体观察和组织学检测其修复效果。实验分3组:(A)转基因细胞片包裹负载有转基因BMSCs的PLGA支架组;(B)负载有转基因BMSCs的PLGA支架组;(C)负载BMSCs的PLGA支架组。结果显示,体外成功收获了完整的GDF5转基因细胞片,Real time PCR检测到GDF5 mRNA的表达,行大体组织的II型胶原免疫组化和阿利新蓝染色显示:A组和B组均表达II型胶原和糖胺聚糖(GAG),但A组表达高于B组,有统计学意义(P<0.05)。由此可得,转基因细胞片包裹负载转基因BMSCs PLGA支架较传统转基因BMSCs负载PLGA支架方法具有更加优越的成软骨能力,能更有效地促进软骨缺损的修复。
文摘目的研究利用骨髓基质细胞膜片复合聚乳乙醇酸（ploy of lactic-co-glycolic acid，PLGA）支撑体，在生物反应器条件下体外构建管状软骨的可行性。方法分离兔骨髓基质细胞，高密度连续培养，转化生长因子-1诱导构建成干细胞膜片，制作圆柱状PLGA支撑体，将细胞膜片均匀缠绕在表面。静置孵育14d，使细胞膜片与PLGA相互贴附后，进入生物反应器动态培养8周后，取出标本。从大体形态、组织学结构、蛋白多糖含量以及生物力学性能等方面评价形成软骨的理化特性。结果通过此策略构建的软骨外观与天然软骨组织非常相似，保持着良好的管状外形，颜色呈乳白色，有光泽，质地均匀，弹性好，具有中等偏软的硬度。组织学结果显示总体结构呈现软骨样结构，HE染色可见软骨样细胞分布于细胞陷窝之中，周围是均匀的细胞外基质，番红-O染色可见细胞外基质着色为鲜红色，提示蛋白多糖含量丰富，有大量软骨基质物产生。结论细胞膜片复合支撑体策略能形成管状形态的软骨组织，为气管软骨的再造提供了新的方法，有可能解决气管缺损的临床难题。
