Control of self-renewal and differentiation of human ES cells (hESCs) remains a challenge. This is largely due to the use of culture systems that involve poorly defined animal products and do not mimic the normal developmental milieu. Routine protocols involve the propagation of hESCs on mouse fibroblast or human feeder layers, enzymatic cell removal, and spontaneous differentiation in cultures of embryoid bodies, and each of these steps involves significant variability of culture conditions. We report that a completely synthetic hydrogel matrix can support (i) long-term self-renewal of hESCs in the presence of conditioned medium from mouse embryonic fibroblast feeder layers, and (ii) direct cell differentiation. Hyaluronic acid (HA) hydrogels were selected because of the role of HA in early development and feeder layer cultures of hESCs and the controllability of hydrogel architecture, mechanics, and degradation. When encapsulated in 3D HA hydrogels (but not within other hydrogels or in monolayer cultures on HA), hESCs maintained their undifferentiated state, preserved their normal karyotype, and maintained their full differentiation capacity as indicated by embryoid body formation. Differentiation could be induced within the same hydrogel by simply altering soluble factors. We therefore propose that HA hydrogels, with their developmentally relevant composition and tunable physical properties, provide a unique microenvironment for the self-renewal and differentiation of hESCs.

译文

:控制人类ES细胞(hESCs)的自我更新和分化仍然是一个挑战。这主要是由于使用了涉及定义不明确的动物产品且不模仿正常发育环境的培养系统。常规方案涉及hESC在小鼠成纤维细胞或人饲养层上的繁殖,酶细胞的去除以及胚状体培养物中的自发分化,并且这些步骤中的每一个都涉及培养条件的显着可变性。我们报告完全合成的水凝胶基质可以支持(i)从小鼠胚胎成纤维细胞饲养层的条件培养基存在下hESCs的长期自我更新,和(ii)直接细胞分化。选择透明质酸(HA)水凝胶是因为HA在hESC的早期发育和饲养层培养中的作用以及水凝胶结构,力学和降解的可控性。当封装在3D HA水凝胶中(但不在其他水凝胶中或在HA的单层培养物中)时,hESC保持其未分化状态,保留其正常核型,并保持其完整的分化能力(如胚状体形成所示)。只需改变可溶性因子,即可在同一水凝胶中诱导分化。因此,我们建议HA水凝胶具有与发育相关的组成和可调节的物理特性,可为hESCs的自我更新和分化提供独特的微环境。

+1
+2
100研值 100研值 ¥99课程
检索文献一次
下载文献一次

去下载>

成功解锁2个技能,为你点赞

《SCI写作十大必备语法》
解决你的SCI语法难题!

技能熟练度+1

视频课《玩转文献检索》
让你成为检索达人!

恭喜完成新手挑战

手机微信扫一扫,添加好友领取

免费领《Endnote文献管理工具+教程》

微信扫码, 免费领取

手机登录

获取验证码
登录