小鼠类胚胎干细胞分离培养及用于制作嵌合体小鼠的实验研究

Experiment on Isolation of Embryonic Stem-like Cells and Generation of Chimerical Mice

目的探讨分离小鼠囊胚内细胞群类胚胎干细胞及用于制作嵌合体小鼠的方法及应用价值。方法分离3.5d小鼠囊胚内细胞群的类胚胎干细胞作为供体细胞,通过显微注射方法将分离的类胚胎干细胞注射到供体小鼠的囊胚腔中,再将注射后的囊胚移植到假孕雌鼠的子宫中制作嵌合体小鼠。结果分离36枚囊胚的内细胞群类胚胎干细胞,注射256只昆明小鼠囊胚中,移植32只假孕雌鼠子宫中,获产崽2窝,共12只,其中2只获毛色嵌合体小鼠。结论采用该技术分离所获得的类胚胎干细胞作为供体细胞制作嵌合体小鼠获得成功,该方法为ES细胞介导的转基因动物制作增添了一条新的途径,在同种不同品系的动物改良及遗传病基因治疗中有一定的应用价值,尤其是对未能建立ES细胞系的大动物的遗传工程操作具有一定意义。

Objective To explore techniques of isolating embryonic stem-like cells (ESLCs) from mouse blastocyst inner cell mass and the utilization of ESLCs to generate chimerical mice. Methods\ ESLCs from mouse blastocyst inner cell masses of 3\^5 days were isolated as donor' cells to inject receptors'blastocysts to generate chimerical mice by micromanipulating techniques, and all those blastocysts were transfered to the uterine of pseudopregnant female mice to gernerate chimerical mice. Results All the ESLCs isolated from 36 inner cell masses were injected into 256 blastocysts of Kunming mice ,and those blastocysts were transferred to uterine of 32 pseudopregnant female mice, 2 chimerical mice from 12 gernerations were obtained. Conclusion \ Technique of isolating ESLCs was a new approach in generating chimerical animals, especially in some animals in which Esc lines were not constructed.