Embryoid body culture of mouse embryonic stem cells using microwell and micropatterned chips
Embryoid body culture of mouse embryonic stem cells using microwell and micropatterned chips
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DOI：
10.1016/j.jbiosc.2010.08.014
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发表时间：
2011-01-01
影响因子：
2.8
通讯作者：
Nakazawa, Kohji
中科院分区：
文献类型：
--
作者：
Sakai, Yusuke;Yoshiura, Yukiko;Nakazawa, Kohji
比较了两种微芯片条件下小鼠胚胎干细胞拟胚体（EB）的增殖和分化特性。微孔芯片在聚甲基丙烯酸甲酯板上含有270个微孔（直径，600 μ m;深度，600 μ m），并且用聚乙二醇（PEG）进行表面改性以使其不具有粘性。微图案化芯片含有270个明胶点（直径，200 μ m）作为玻璃板上的细胞粘附区域;将缺少这些点的区域PEG修饰以使其不粘附。ES细胞自发地从芯片中每个微孔中的细胞聚集体形成EB。相反，接种到图案化芯片上的细胞在明胶点上形成单层，并逐渐增殖形成EB。图案化芯片中的EB保持高细胞生长速率，并且内胚层（TTR和AFP）和中胚层（Nkx2.5、α MHC、Flk 1和PDGFR β）标志物的表达增加，并且这些细胞性质与先前的方法（悬滴和圆底96孔板培养物）相似。相比之下，微孔芯片中ES细胞的增殖低于图案化芯片和先前的方法，并且EB分化进行缓慢并且仅形成少量内胚层。这些结果表明，胚胎干细胞在微芯片培养过程中产生EB的过程不同，可能会影响胚胎干细胞的增殖和分化，微芯片中微孔结构的存在会抑制胚胎干细胞的增殖和分化进程。(C)2010年，生物技术学会，日本。All rights reserved.
The proliferation and differentiation properties of embryoid bodies (EB) from mouse embryonic stem (ES) cells were compared under two microchip conditions: microwell chip and micropatterned chip. The microwell chip contained 270 microwells (diameter, 600 mu m; depth, 600 mu m) on a polymethylmethacrylate plate and was surface-modified with polyethylene glycol (PEG) to render it non-adhesive. The micropatterned chip contained 270 gelatin spots (diameter, 200 mu m) as the cell adhesion area on a glass plate; the region lacking these spots was PEG-modified to render it non-adhesive. The ES cells spontaneously formed the EBs from cell aggregates in each microwell in the chip. In contrast, cells inoculated onto the patterned chip formed a monolayer on the gelatin spots and gradually proliferated to form EBs. The EBs in the patterned chip maintained the high cell growth rate and the expression of endoderm (TTR and AFP) and mesoderm (Nkx2.5, alpha MHC, Flk1, and PDGFR beta) markers was increased, and these cell properties were similar to the previous methods (hanging drop and round-bottomed 96-well plate cultures). In contrast, the proliferation of ES cells in the microwell chip was lower than in the patterned chip and previous methods, and the EB differentiation proceeded slowly and only formed a small amount of endoderm. These results indicate that the difference of EB generating process in the microchip cultures may affect to the proliferation and differentiation of ES cells, and the existence of microwell structure in the microchip downregulates the cell proliferation and the differentiated progress of ES cells. (C) 2010, The Society for Biotechnology, Japan. All rights reserved.