人羊水幹細胞（hAFSCs）是多能胚胎細胞，它能夠分化成多個譜系。分離的hAFSCs貼附和增殖取決於所使用的基質。微環境在hAFSCs的分化和基因表達中扮演著關鍵的角色。因此，在本研究中，我們設計了hAFSCs的培養方法，其中將不同的基質（細胞外間質或熱敏感聚合物）塗覆被在組織培養聚苯乙烯（TCPS）培養皿上，可以在成骨細胞和軟骨細胞獲得更高的細胞增殖和分化能力。我們培養了使用不同基質的hAFSCs，例如TCPS以及TCPS塗有（a）基質膠，（b）Synthemax II，（c）人類重組-玻璃粘連蛋白（rVN），（d）第一型膠原蛋白，（e）纖連蛋白（f）聚（N-異丙基丙烯酰胺）（PolyNIPAAm），（g）聚（N-異丙基丙烯酰胺-丙烯酸丁酯共聚物）（PolyNIPAAm-BA）和（h）用ECM固定的熱敏感聚合物（NIPAAm-ECMs）。 在這些材料中，在rVN，Synthemax II和NIPAAm-ECM上培養的hAFSCs呈現出更高的增殖和分化能力。 此外，我們研究了使用間充質幹細胞（MSCs）衍生為視網膜色素上皮細胞（RPE）的潛力和可行性。先前的研究已經使用人類胚胎幹細胞（hESC）或人類誘導性多能幹細胞（hiPSC）來分化成RPE。然而，這可能導致免疫排斥和畸胎瘤產生的問題。對於使用間充質乾細胞（MSCs），例如hAFSC衍生的自RPE，應該更具前瞻性。 而且我們開發了一種新的細胞培養基，它使用人血小板裂解液（hPL）代替胎牛血清（FBS）作為補充劑，以提高hAFSCs的分化率，並建立更完整的hAFSCs無異種培養條件作為未來的臨床應用。 ;Human amniotic fluid stem cells (hAFSCs) are pluripotent fetal cells, which are capable to differentiate into multiple lineages. The isolated hAFSCs adhesion and proliferation depending on the substrates used. Microenvironment plays a key role in differentiation and gene expression for hAFSCs. Therefore, in this study, we designed a culture method of hAFSCs where the different substrates (extracellular matrices or thermoresponsive polymers) were coated on tissue culture polystyrene (TCPS) dishes could get higher cell proliferation and differentiation ability into osteoblasts and chondrocytes. We cultivated the hAFSCs where on different substrates were used such as TCPS and TCPS coated with (a) Matrigel, (b) Synthemax II, (c) human recombinant-vitronectin (rVN), (d) collagen type I, (e) fibronectin, (f) poly(N-isopropylacrylamide) (PNIPAAm), (g) poly(N-isopropylacrylamide-co-butylacrylate) (PNIPAAm-BA) and (h) thermoresponsive polymers immobilized with ECMs (PNIPAAm-ECMs). Among these materials hAFSCs cultured on rVN, Synthemax II and PNIPAAm-ECMs presented the higher proliferation and differentiation abilities. Moreover, we investigated the potential and feasibility of mesenchymal stem cells (MSCs) derived into retinal pigment epithelium (RPE). Previous studies have used human embryonic stem cells (hESCs) or human induced pluripotent stem cells (hiPSCs) to differentiate into RPE. However, this may cause immune rejection and teratoma production problems. It should be more prospective for mesenchymal stem cells (MSCs) such as hAFSCs derived into RPE. Furthermore, a new cell culture medium was developed, which used human platelet lysate (hPL) instead of fetal bovine serum (FBS) as a supplement to improve the differentiation ratio of hAFSCs and built up a more complete hAFSCs with xeno-free culture conditions for clinical application in future.
