The microwell control of embryoid body size in order to regulate cardiac differentiation of human embryonic stem cells.
The microwell control of embryoid body size in order to regulate cardiac differentiation of human embryonic stem cells.
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DOI：
10.1016/j.biomaterials.2009.11.033
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发表时间：
2010-03
期刊：
影响因子：
14
通讯作者：
Kamp, Timothy J.
中科院分区：
文献类型：
--
作者：
Mohr, Jeffrey C.;Zhang, Jianhua;Azarin, Samira M.;Soerens, Andrew G.;de Pablo, Juan J.;Thomson, James A.;Lyons, Gary E.;Palecek, Sean P.;Kamp, Timothy J.
利用胚胎样体（EBs）将人胚胎干细胞（hESCs）分化为心肌细胞（CMs）的效率相对较低且变化很大。使用标准的酶解技术形成EBs，产生各种大小和几何形状的EBs。利用3-D立方体微井系统，在横向尺寸为100 ~ 500 μm、深度为120 μm的特定尺寸集落中培养hESCs，可以形成更均匀尺寸的EBs。300 μm微孔产生的EBs收缩率最高，但对表达肌球蛋白轻链2A （MLC2a）的细胞的流式细胞术显示，100 μm微孔和300 μm微孔在EBs中形成的心肌细胞百分比相似（约3%）。这些数据以及抗mf20和MLC2a的免疫标记表明，较小的EB不太可能形成收缩的EBs，但与较大的EB相比，这些收缩的EBs在心肌细胞中相对丰富，其中CMs占总细胞的比例较小。我们得出结论，微孔工程EB大小调节心脏发生，可用于更有效和可重复的hESC-CMs的形成，用于研究和治疗应用。
The differentiation of human embryonic stem cells (hESCs) into cardiomyocytes (CMs) using embryoid bodies (EBs) is relatively inefficient and highly variable. Formation of EBs using standard enzymatic disaggregation techniques results in a wide range of sizes and geometries of EBs. Use of a 3-D cuboidal microwell system to culture hESCs in colonies of defined dimensions, 100 to 500 μm in lateral dimensions and 120 μm in depth, enabled formation of more uniform sized EBs. The 300 μm microwells produced highest percentage of contracting EBs, but flow cytometry for myosin light chain 2A (MLC2a) expressing cells revealed a similar percentage (~3%) of cardiomyocytes formed in EBs from 100 μm and 300 μm microwells. These data, and immunolabeling with anti-MF20 and MLC2a, suggest that the smaller EBs are less likely to form contracting EBs, but those contracting EBs are relatively enriched in cardiomyocytes compared to larger EB sizes where CMs make up a proportionately smaller fraction of the total cells. We conclude that microwell-engineered EB size regulates cardiogenesis and can be used for more efficient and reproducible formation of hESC-CMs needed for research and therapeutic applications.
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影响因子：
14
作者：
Mohr, Jeffrey C.;de Pablo, Juan J.;Palecek, Sean P.
通讯作者：
Palecek, Sean P.
影响因子：
20.1
作者：
Rudy-Reil, D;Lough, J
通讯作者：
Lough, J
影响因子：
20.3
作者：
Ng, ES;Davis, RP;Elefanty, AG
通讯作者：
Elefanty, AG
影响因子：
15.9
作者：
Kehat, I;Kenyagin-Karsenti, D;Gepstein, L
通讯作者：
Gepstein, L
影响因子：
37.8
作者：
Mummery, C;Ward-van Oostwaard, D;Tertoolen, L
通讯作者：
Tertoolen, L