Kang Sun-Woong, Seo Sang-Woo, Choi Cha Yong, Kim Byung-Soo
Department of Chemical Engineering, Hanyang University, Seoul, Korea.
Tissue Eng Part C Methods. 2008 Mar;14(1):25-34. doi: 10.1089/tec.2007.0290.
Tissue engineering often requires ex vivo cell expansion to obtain a large number of transplantable cells. However, the trypsinization process used to harvest ex vivo expanded cells for transplantation interrupts interactions between cultured cells and their extracellular matrices, facilitating apoptosis and consequently limiting the therapeutic efficacy of the transplanted cells. In the present study, open macroporous poly(lactic-co-glycolic acid) (PLGA) microspheres were used as a cell culture substrate to expand human adipose-derived stromal cells (ASCs) ex vivo and as a cell transplantation vehicle for adipose tissue engineering, thus avoiding the trypsinization necessary for transplantation of ex vivo expanded cells. Human ASCs cultured on macroporous PLGA microspheres in stirred suspension bioreactors expanded 3.8-fold over 7 days and differentiated into an adipogenic lineage. The apoptotic activity of ASCs cultured on microspheres was significantly lower than that of trypsinized ASCs. ASCs cultured on microspheres survived much better than trypsinized ASCs upon transplantation. The implantation of ASCs cultured on microspheres resulted in much more extensive adipose tissue formation than the implantation of ASCs cultured on plates, trypsinized, and subsequently mixed with microspheres. Ex vivo cell expansion and transplantation using this system would improve the therapeutic efficacy of cells over the current methods used for tissue engineering.
组织工程通常需要体外细胞扩增以获得大量可移植细胞。然而,用于收获体外扩增细胞进行移植的胰蛋白酶消化过程会中断培养细胞与其细胞外基质之间的相互作用,促进细胞凋亡,从而限制移植细胞的治疗效果。在本研究中,开放大孔聚乳酸-乙醇酸共聚物(PLGA)微球被用作细胞培养底物,用于体外扩增人脂肪来源的基质细胞(ASC),并作为脂肪组织工程的细胞移植载体,从而避免了体外扩增细胞移植所需的胰蛋白酶消化过程。在搅拌悬浮生物反应器中,在大孔PLGA微球上培养的人ASC在7天内扩增了3.8倍,并分化为脂肪生成谱系。在微球上培养的ASC的凋亡活性明显低于经胰蛋白酶处理的ASC。移植后,在微球上培养的ASC比经胰蛋白酶处理的ASC存活得好得多。与在平板上培养、经胰蛋白酶处理并随后与微球混合的ASC植入相比,植入在微球上培养的ASC导致形成更广泛得多的脂肪组织。使用该系统进行体外细胞扩增和移植将比目前用于组织工程的方法提高细胞的治疗效果。