中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2006年
1期
7-11
,共5页
张嘉晴%余伟华%张秀明%彭延文%李伟强%陈蕊%余新炳%李树浓%项鹏
張嘉晴%餘偉華%張秀明%彭延文%李偉彊%陳蕊%餘新炳%李樹濃%項鵬
장가청%여위화%장수명%팽연문%리위강%진예%여신병%리수농%항붕
小鼠%胚胎干细胞%杂交
小鼠%胚胎榦細胞%雜交
소서%배태간세포%잡교
Mice%Embryonic stem cells%Hybridization
目的:建立C57BL/6J×129/J杂交小鼠ES细胞系.方法:收集3.5 d.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.3-4 d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.
目的:建立C57BL/6J×129/J雜交小鼠ES細胞繫.方法:收集3.5 d.p.c.的囊胚,培養在預先鋪有小鼠成纖維細胞(MEFs)的高糖DMEM培養液中.3-4 d後,挑齣內細胞糰(ICM),消化後重新種到新鮮的有MEFS培養液中.等到有典型的ES樣集落長齣,即傳代以得到永久ES細胞繫.通過分析堿性燐痠酶活性,SSEA-1,Oct-4的錶達和形成畸胎瘤的能力來鑒定ES細胞的多嚮分化能力.結果:穫得的兩箇C57BL/6J×129/J雜交小鼠ES細胞繫絕大多數細胞具有正常的覈型(40,XY),堿性燐痠酶染色暘性,SSEA-1,Oct-4錶達暘性,ES細胞註入SCID鼠後可穫得來自3箇胚層的組織.結論:建立瞭兩株具有長期自我更新能力和多嚮分化潛能的C57BL/6J×129/J雜交小鼠ES細胞繫.
목적:건립C57BL/6J×129/J잡교소서ES세포계.방법:수집3.5 d.p.c.적낭배,배양재예선포유소서성섬유세포(MEFs)적고당DMEM배양액중.3-4 d후,도출내세포단(ICM),소화후중신충도신선적유MEFS배양액중.등도유전형적ES양집락장출,즉전대이득도영구ES세포계.통과분석감성린산매활성,SSEA-1,Oct-4적표체화형성기태류적능력래감정ES세포적다향분화능력.결과:획득적량개C57BL/6J×129/J잡교소서ES세포계절대다수세포구유정상적핵형(40,XY),감성린산매염색양성,SSEA-1,Oct-4표체양성,ES세포주입SCID서후가획득래자3개배층적조직.결론:건립료량주구유장기자아경신능력화다향분화잠능적C57BL/6J×129/J잡교소서ES세포계.
AIM: To establish hybrid mouse embryonic stem (ES) 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 ES - like colonies appeared, passaged them to give permanent ES cell lines. The pluripotent propertes of ES 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 ES cell lines, SC1001, SC1002 were obtained from blastocysts of the (C57BL/6J × 129/J) F1 genotype. Most of these ES 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 ES cell lines having pluripotent properties and capacity for long - term self renewal were generated from blastocysts of the ( C57BL/6J × 129/J) F1 genotype.
