Forced aggregation of defined numbers of human embryonic stem cells into embryoid bodies fosters robust, reproducible hematopoietic differentiation
Forced aggregation of defined numbers of human embryonic stem cells into embryoid bodies fosters robust, reproducible hematopoietic differentiation
复制标题
DOI：
10.1182/blood-2005-03-0987
复制
发表时间：
2005-09-01
期刊：
影响因子：
20.3
通讯作者：
Elefanty, AG
中科院分区：
文献类型：
--
作者：
Ng, ES;Davis, RP;Elefanty, AG
为了实现人类胚胎干细胞（hESC）的治疗潜力，有必要以统一和可重复的方式调节其分化。我们已经开发了一种方法，其中已知数量的hESC在无血清培养基中通过离心聚集以促进形成大小均匀的胚状体（EB）（自旋EB）。这些自旋EB分化有效和同步，证明了代表干细胞，原条，和中胚层的分子标记物的顺序表达。在造血生长因子的存在下，实现了可重复的分化，在超过90%的EB中形成了血细胞。在克隆形成测定中使用从绿色荧光蛋白阳性（GFP（+））和GFP（-）hESC的混合物产生的嵌合EB，造血前体频率估计为约1：500输入细胞。这种EB形成方法提供了一种普遍适用的调节和客观监测hESC定向分化的方法。
To realize the therapeutic potential of human embryonic stem cells (hESCs), it is necessary to regulate their differentiation in a uniform and reproducible manner. We have developed a method in which known numbers of hESCs in serum-free medium were aggregated by centrifugation to foster the formation of embryoid bodies (EBs) of uniform size (spin EBs). These spin EBs differentiated efficiently and synchronously, as evidenced by the sequential expression of molecular markers representing stem cells, primitive streak, and mesoderm. In the presence of hematopoietic growth factors, reproducible differentiation was achieved with blood cells formed in more than 90% of EBs. Using chimeric EBs generated from mixtures of green fluorescence protein-positive (GFP(+)) and GFP(-) hESCs in a clonogenic assay, hematopoietic precursor frequency was estimated to be approximately 1:500 input cells. This method of EB formation provides a generally applicable means for modulating and objectively monitoring the directed differentiation of hESCs.