細胞培養基材對3D培養幹細胞球的影響探討

Abstract

間葉幹細胞(mesenchymal stem cells，MSCs)是組織工程中經常使用的細胞，文獻中指出，藉由培養基材，可以達成分離幹細胞、維持幹細胞特性等目標，可見培養基材之重要性。然而細胞培養基材在功能性上尚有很大的研究空間，在此我們利用了DNA質體 pTRE-Tight-DsRED2 (3.3 kb，~5 nm)展示了藉由細胞培養基材(hyaluronan-modified chitosan，chitosan-HA)進行3D培養，能夠達成原本無法達成的不使用轉殖劑的幹細胞基因轉殖，以及在不同基材上(chitosan和chitosan-HA)進行MSCs和血管內皮前驅細胞(endothelial progenitor cells，EPCs)共培養時，能夠藉由改變培養基材和細胞比例達成不同型態的自組裝混和細胞球，而動物實驗暗示了自組裝型態對於血管新生有極大潛力。

Mesenchymal stem cells, MSCs are the cell often used in Tissue engineering. Prior literature shows stem cells isolation and maintain the stemness can be chieved by the cell culture substrate. This illustrate the improtance of the cell culture substrate. However, the funtionilties of cell culture substrate need futher research. Here we use plsmid DNA pTRE-Tight-DsRED2 (3.3 kb, ~5 nm) and cultue stem cell spheroids on hyaluronan-modified chitosan, chitosan-HA to demonstrate a substrate mediated stem cell gene transfect without transfect reagent which been consider unlikly happend by. On the other hand, we observed different self-assembled morphologies (i.e. randomly mixed or concentric spheroids) when co-cultured MSCs and endothelial progenitor cells, EPCs by changed the initial cell ratio or replaced the substrate chitosan or chitosan-HA. In particular, the self-assembled MSC-EPC core-shell concentric spheroids show a greater angiogenic effect in vitro.