Research on manipulation of mammalian fetal germ cells and gene transfection
哺乳动物胎儿生殖细胞操作及基因转染研究
基本信息
- 批准号:07558290
- 负责人:
- 金额:$ 4.1万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (A)
- 财政年份:1995
- 资助国家:日本
- 起止时间:1995 至 1997
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
To develop novel methods of reproductive and genetical manipulation of mammalian fetal germ cells, we carried out the in vitro culture, gene transfection, and micromanipulation of mouse fetal germ cells. Firstly, we improved culture conditions of primordial germ cells. We then tested various methods of gene transfection into cultured fetal germ cells. We observed inhibition of apoptosis by transfection of bcl-x gene and adenovirus E1b 19k gene. We also tested differentiation capacity of EG cells established by addition of forskolin and retinoic acid in culture of primordial germ cells. After transplantation into the W mutant mouse testis, they showed differentiation capacity of ES or teratoma cells but not the germ cells.Also, new micromanipulation methods were developed to produce offspring from germ cells in various artificial fertilization procedures. We first improved the method of using round spermatid for production of fertilized eggs by micromanipulation. We also succeeded to produce normal diploid embryos by using primary spermatocytes and ovum. During the microsurgical fertilization using spermatid, a GFP gene solution was injected at the same time. Such developed blastocytes showed expression of the GFP fluorescence, thus indicating possibility of the gene transfection using such procedure.
为了开发哺乳动物胎儿生殖细胞的生殖和遗传操作的新方法,我们对小鼠胎儿生殖细胞进行了体外培养、基因转染和显微操作。首先,我们改善了原始生殖细胞的培养条件。然后我们测试了将基因转染到培养的胎儿生殖细胞中的各种方法。我们观察到转染bcl-x基因和腺病毒E1b 19k基因对细胞凋亡的抑制。我们还测试了在原始生殖细胞培养物中添加毛喉素和视黄酸建立的EG细胞的分化能力。移植到W突变小鼠睾丸后,它们显示出ES或畸胎瘤细胞的分化能力,但不显示生殖细胞的分化能力。此外,还开发了新的显微操作方法,以在各种人工受精过程中从生殖细胞产生后代。我们首先改进了通过显微操作利用圆形精子细胞产生受精卵的方法。我们还利用原代精母细胞和卵子成功地产生了正常的二倍体胚胎。在使用精子细胞进行显微外科受精过程中,同时注射GFP基因溶液。这种发育的胚细胞显示出GFP荧光的表达,从而表明使用这种程序进行基因转染的可能性。
项目成果
期刊论文数量(31)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Ogura A, Wakayama T, Suzuki O, Shin T-Y, Matsuda J, Kobayashi Y.: "Chromosomes of mouse primary spermatocytes undergo meiotic divisions after incorporation into homologous immature oocytes." Zygote. 5. 177-182 (1997)
Ogura A、Wakayama T、Suzuki O、Shin T-Y、Matsuda J、Kobayashi Y.:“小鼠初级精母细胞的染色体在并入同源未成熟卵母细胞后经历减数分裂。”
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- 影响因子:0
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Nohara,M: "Development of a successful in vitro fertilization procedure and partial characterization of the gamete in the Mastomys(Praomys coucha):a new species for laboratory research in reproductive biology" Biol.Reprod. 58. 226-233 (1998)
Nohara,M:“成功的体外受精程序的开发和 Mastomys(Praomys coucha)配子的部分表征:生殖生物学实验室研究的新物种”Biol.Reprod。
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Koshimizu,U.,他: "Functional requirement of gp 130-mediated signaling for growth and survival of mouse primordial germ cells in vitro and derivation of embryonic germ (EG) cells." Development. 122. 1235-1242 (1996)
Koshimizu, U., et al.:“gp 130 介导的信号对小鼠原始生殖细胞体外生长和存活以及胚胎生殖 (EG) 细胞的衍生的功能要求。” 122. 1235-1242 (1996)。
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Wakayam T,Hayashi Y,Ogura A.: "Participation of the female pronucleus derived from the second polar body in full embryonic development of mice." J.Reprod.Fertil.110. 263-266 (1997)
Wakayam T、Hayashi Y、Ogura A.:“来自第二极体的雌性原核参与小鼠的完整胚胎发育。”
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- 期刊:
- 影响因子:0
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- 通讯作者:
Ogura,A: "Chromosomes of mouse primary spermatocytes undergo meiotic divisions after incorporation into homologous immature oocytes" Zygote. 5. 177-182 (1997)
Ogura,A:“小鼠初级精母细胞的染色体在并入同源未成熟卵母细胞后经历减数分裂”合子。
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NAKATSUJI Norio的其他文献
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{{ truncateString('NAKATSUJI Norio', 18)}}的其他基金
Establishment of a isolation and maturation system for human adult hepatic stem cells
人成体肝干细胞分离成熟体系的建立
- 批准号:
14370384 - 财政年份:2002
- 资助金额:
$ 4.1万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Regulative Mechanisms and Manipulation of the Germ Cell Lineage
生殖细胞谱系的调节机制和操纵
- 批准号:
11234101 - 财政年份:1999
- 资助金额:
$ 4.1万 - 项目类别:
Grant-in-Aid for Scientific Research on Priority Areas
Research on Development and Sex-Differentiation of Mouse Germ Cells
小鼠生殖细胞发育及性别分化的研究
- 批准号:
11480223 - 财政年份:1999
- 资助金额:
$ 4.1万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Research on development and sex differentiation of mouse fetal germ cells
小鼠胎儿生殖细胞发育及性别分化的研究
- 批准号:
11234201 - 财政年份:1999
- 资助金额:
$ 4.1万 - 项目类别:
Grant-in-Aid for Scientific Research on Priority Areas
Research on regulation mechanisms of development and differentiation of mouse primordial germ cells
小鼠原始生殖细胞发育分化调控机制研究
- 批准号:
08458241 - 财政年份:1996
- 资助金额:
$ 4.1万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Research on control mechanisms of growth and differentiation of mouse primordial germ cells and their developmental manipulation
小鼠原始生殖细胞生长分化调控机制及其发育调控研究
- 批准号:
05454655 - 财政年份:1993
- 资助金额:
$ 4.1万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
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