C57BL/6J×129/J杂交小鼠ES细胞系的建株(英文)

Establishment and characteristics of hybrid embryonic stem cell lines from blastocysts of the (C57BL/6J×129/J)F1 mouse

目的:建立C57BL/6J×129/J杂交小鼠ES细胞系。方法:收集3.5d.p.c.的囊胚,培养在预先铺有小鼠成纤维细胞(MEFs)的高糖DMEM培养液中。3-4d后,挑出内细胞团(ICM),消化后重新种到新鲜的有MEFS培养液中。等到有典型的ES样集落长出,即传代以得到永久ES细胞系。通过分析碱性磷酸酶活性,SSEA-1,Oct-4的表达和形成畸胎瘤的能力来鉴定ES细胞的多向分化能力。结果:获得的两个C57BL/6J×129/J杂交小鼠ES细胞系绝大多数细胞具有正常的核型(40,XY),碱性磷酸酶染色阳性,SSEA-1,Oct-4表达阳性,ES细胞注入SCID鼠后可获得来自3个胚层的组织。结论:建立了两株具有长期自我更新能力和多向分化潜能的C57BL/6J×129/J杂交小鼠ES细胞系。

AIM： To establish hybrid mouse embryonic stem （KS） cell line from blastocysts of the （C57BL/6J × 129/ J） F1 mouse. METHODS：3.5 days post- coitus （d. p.c. ） blastocysts were cultured on mouse embryonic fibroblasts （MEFs） in the medium, after 3 - 4 days, Inner cell mass were picked up and disaggregated, then reseeded. After the KS - like colonies appeared, passaged them to give permanent KS cell lines. The pluripotent properties of KS cells obtained were analyzed by alkaline phosphatase （AKP） activity, expression of SSEA- 1 and Oct- 4, and their capacity to form teratoma. RESULTS： Two hybrid KS cell lines, SC1001, SC1002 were obtained from blastocysts of the （C57BL/6J × 129/J） F1 genotype. Most of these KS cells had a normal karyotype and an XY sex chromosome composition. The pluripotent properties of the cell lines were analyzed on the basis of their alkaline phosphatase activity, expression of SSEA - 1 and Oct - 4, and their capacity to form teratoma in severe combined immunodeficiency （SCID） mice. CONCLUSION： Two hybrid mouse KS cell lines having pluripotent properties and capacity for long - term self renewal were generated from blastocysts of the （C57BL/6J × 129/J） F1 genotype.