Division of Matrix Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden.
Nat Biotechnol. 2010 Jun;28(6):611-5. doi: 10.1038/nbt.1620. Epub 2010 May 30.
We describe a system for culturing human embryonic stem (hES) cells and induced pluripotent stem (iPS) cells on a recombinant form of human laminin-511, a component of the natural hES cell niche. The system is devoid of animal products and feeder cells and contains only one undefined component, human albumin. The hES cells self-renewed with normal karyotype for at least 4 months (20 passages), after which the cells could produce teratomas containing cell lineages of all three germ layers. When plated on laminin-511 in small clumps, hES cells spread out in a monolayer, maintaining cellular homogeneity with approximately 97% OCT4-positive cells. Adhesion of hES cells was dependent on alpha6beta1 integrin. The use of homogeneous monolayer hES or iPS cell cultures provides more controllable conditions for the design of differentiation methods. This xeno-free and feeder-free system may be useful for the development of cell lineages for therapeutic purposes.
我们描述了一种在重组人层粘连蛋白-511(一种天然 hES 细胞龛的组成部分)上培养人胚胎干细胞(hES)和诱导多能干细胞(iPS)的系统。该系统不含动物产品和饲养细胞,仅含有一种未定义的成分,即人白蛋白。hES 细胞具有正常核型,至少自我更新了 4 个月(20 代),之后这些细胞可以产生含有三个胚层细胞谱系的畸胎瘤。当在小团块上接种到层粘连蛋白-511 上时,hES 细胞会在单层中展开,大约 97%的 OCT4 阳性细胞保持细胞同质性。hES 细胞的黏附依赖于 alpha6beta1 整联蛋白。使用同质单层 hES 或 iPS 细胞培养物可为分化方法的设计提供更可控的条件。这种无动物成分和无饲养细胞的系统可能对治疗目的的细胞谱系的开发有用。