Method for neural differentiation of embryonic stem cells using protease passaging techniques
申请人:Schulz C. Thomas
公开号:US20060183221A1
公开(公告)日:2006-08-17
The present invention provides methods for human pluripotent cell culturing and for neural cell production. More particularly, the present invention provides culturing methods employing dissociating cell cultures to an essentially single cell culture, such as by employing antibody selection and bulk passaging treatments utilizing the subsequent application of Collagenase and trypsin. In certain embodiments, the cells are further treated with essentially serum free MEDII conditioned medium, proline, or minimal medium, and are optionally treated with amphiphilic lipid compounds for the generation of human neural cells from pluripotent human cells. In certain embodiments, the cells cultured using these methods have an abnormal karyotype.
本发明提供了人类多能细胞培养和神经细胞生产的方法。更具体地说,本发明提供了将细胞培养物解离成基本单细胞培养物的培养方法,例如通过抗体选择和随后应用胶原酶和胰蛋白酶的批量传代处理。在某些实施方案中,细胞进一步用基本无血清的 MEDII 条件培养基、脯氨酸或最低限度培养基处理,并可选择用两亲脂质化合物处理,以便从多能人类细胞中生成人类神经细胞。在某些实施方案中,使用这些方法培养的细胞具有异常核型。