Development of Pluripotential Embryonic Stem Cells from Rat Blastocyst

大鼠囊胚多能胚胎干细胞的发育

• 批准号: 07558238
• 负责人: SUGIYAMA Fumihiro
• 金额: $ 0.38万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07558238/
• 关键词: Embryonic stem cells; IGF-II; LIF; Rats; ES細胞; キメラ

To establish pluripotential diploid cells from rat embryos, we cultured blastocysts in medium containing mouse leukemia inhibiting factor (LIF) and insulin-like growth factor (IGF) -II.Combination of mouse LIF (5,000 units/ml) and rat IGF-II (100ng/ml) promoted the growth of inner cell mass (ICM) of rat blastocyst. The form of the cells derived from ICM was similar to that of mouse embryonic stem (ES) cell. We established 9 and 3 lines on ACI and a hybrid (ACI * Wistar-Imamichi) rat, respectively. For evaluating in vivo pluripotency of the rat ES-like cells, 8-cell embryos of Wistar-Imamichi were co-cultured with the established cells to make chimeric rats. On the cell lines derived from ACI rat, 25 neonates were obtained from 349 chimeric embryos that implanted into pseudopregnancy rat. On the neonates, no chimeric rat was observed. By microinjection method, chimeric embryos were produced between the established cells and blastocysts of Wistar-Imamichi. On the cell lines derived from both ACI and the hyberid rats, 304 chimeric embryos were produced. Chimeric rats were not, however, founded in 206 neonates. These findings suggest that, in addition to LIF and IGF-II,other factor may be needed to establish pluripotential ES cells from rat embryos.

为了建立大鼠胚胎多能二倍体细胞，我们将大鼠囊胚培养在含有小鼠白血病抑制因子（LIF）和胰岛素样生长因子（IGF）-Ⅱ的培养基中。小鼠LIF（5，000单位/ml）和大鼠IGF-Ⅱ（100 ng/ml）的联合作用促进了大鼠囊胚内细胞团（ICM）的生长。ICM来源的细胞形态与小鼠胚胎干细胞相似。我们建立了9和3线ACI和杂交（ACI * Wistar-Imamichi）大鼠，分别。为了评价大鼠ES样细胞的体内多能性，将Wistar-Imamichi的8细胞胚胎与建立的细胞共培养以制备嵌合大鼠。在ACI大鼠细胞系上，从349个嵌合体胚胎植入假孕大鼠体内获得25个新生儿。在新生大鼠中，未观察到嵌合体大鼠。通过显微注射法，将已建立的细胞与Wistar-Imamichi囊胚进行嵌合。在来自ACI和hyberid大鼠的细胞系上，产生了304个嵌合体胚胎。然而，在206只新生儿中未发现嵌合体大鼠。这些发现表明，除了LIF和IGF-II，其他因素可能需要建立多能ES细胞从大鼠胚胎。

项目成果

Sugiyama F., Fukamizu A., Miyazaki H., Yagami K., and Murakami K.: "Developmental mechanism in organ disorder combined with hypertension (Japanese)" BIO Clinica. 11. 522-525 (1996)

Sugiyama F.、Fukamizu A.、Miyazaki H.、Yagami K. 和 Murakami K.：“器官疾病合并高血压的发育机制（日语）”BIO Clinica。

Goto Y. et al.: "Evaluation of coculture aggregation for production of germline chimae" Lat.Anim.Sci.45. 601-603 (1995)

Goto Y. 等人：“种系嵌合体生产的共培养聚集的评估”Lat.Anim.Sci.45。

杉山文博: "高血圧とそれに伴う臓器障害の発生機構" BIO Clinica. 11. 522-525 (1996)

Fumihiro Sugiyama：“高血压和相关器官损伤的发生机制”BIO Clinica 11. 522-525 (1996)。

Goto Y., Sugiyama F., Tanimoto K., Ishida J., Syouji M., Takahashi A., Murakami K., Sugiyama Y., Fukamizu A., and Yagami K.: "Evaluation of coculture aggregation for production of germline chimaeras." Lab.Anim.Sci.45. 601-603 (1995)

Goto Y.、Sugiyama F.、Tanimoto K.、Ishida J.、Syouji M.、Takahashi A.、Murakami K.、Sugiyama Y.、Fukamizu A. 和 Yagami K.：“用于种系生产的共培养聚集的评估

Sugiyama F., Fukamizu A., and Murakami K.: "Transgenic animal. Hypertension (Japanese)" Medical term library. 18-19 (1995)

Sugiyama F.、Fukamizu A. 和 Murakami K.：“转基因动物。高血压（日语）”医学术语库。