Sîrbu-Boeţi Mirela-Patricia, Chivu Mihaela, Pâslaru Liliana Livia, Efrimescu C, Herlea V, Pecheanu C, Moldovan Lucia, Dragomir Laura, Bleotu Coralia, Ciucur Elena, Vidulescu Cristina, Vasilescu Mihaela, Boicea Anişoara, Mănoiu S, Ionescu M I, Popescu I
Fundeni Clinical Institute, Center of General Surgery and Liver Transplantation, Bucharest, Romania.
Chirurgia (Bucur). 2009 Jan-Feb;104(1):55-65.
Transplanted mesenchymal stem cells (MSCs) appear to play a significant role in adult tissue repair. The aim of this research was to obtain MSCs enriched, three dimensional (3D) patches for transplant, and to test their ability to induce repair of iatrogenic digestive tract defects in rats. MSCs were obtained from human and rat bone marrow, cultured in vitro, and seeded in a collagen-agarose scaffold, where they showed enhanced viability and proliferation. The phenotype of the cultured cells was representative for MSCs (CD105+, CD90+, and CD34-, CD45-, CD3-, CD14-). The 3D patch was obtained by laying the MSCs enriched collagen-agarose scaffold on a human or swine aortic fragment. After excision of small portions of the rat digestive tract, the 3D patches were sutured at the edge of the defect using micro-surgical techniques. The rats were sacrificed at time-points and the regeneration of the digestive wall was investigated by immunofluorescence, light and electron microscopy. The MSCs enriched 3D patches were biocompatible, biodegradable, and prompted the regeneration of the four layers of the stomach and intestine wall in rats. Human cells were identified in the rat regenerated digestive wall as a hallmark of the transplanted MSCs. For the first time we constructed 3D patches made of cultured bone marrow MSCs, embedded into a collagen-rich biomatrix, on vascular bio-material support, and transplanted them in order to repair iatrogenic digestive tract defects. The result was a complete repair with preservation of the four layered structure of the digestive wall.
移植的间充质干细胞(MSCs)似乎在成体组织修复中发挥着重要作用。本研究的目的是获得富集MSCs的三维(3D)移植补片,并测试其诱导大鼠医源性消化道缺损修复的能力。从人和大鼠骨髓中获取MSCs,在体外培养,然后接种到胶原-琼脂糖支架中,在该支架中它们显示出增强的活力和增殖能力。培养细胞的表型具有MSCs的代表性(CD105+、CD90+,以及CD34-、CD45-、CD3-、CD14-)。通过将富集MSCs的胶原-琼脂糖支架放置在人或猪的主动脉片段上获得3D补片。切除大鼠消化道的小部分后,使用显微外科技术将3D补片缝合在缺损边缘。在不同时间点处死大鼠,通过免疫荧光、光学和电子显微镜观察消化道壁的再生情况。富集MSCs的3D补片具有生物相容性、可生物降解性,并促进了大鼠胃和肠壁四层结构的再生。在大鼠再生的消化道壁中鉴定出人类细胞,作为移植MSCs的标志。我们首次构建了由培养的骨髓MSCs制成的3D补片,将其嵌入富含胶原的生物基质中,置于血管生物材料支架上,并进行移植以修复医源性消化道缺损。结果是实现了完全修复,并保留了消化道壁的四层结构。