Searching for culture condition optimal for proliferation of spermatogonial stem cells

寻找最适合精原干细胞增殖的培养条件

基本信息

  • 批准号:
    15591702
  • 负责人:
  • 金额:
    $ 1.34万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
  • 财政年份:
    2003
  • 资助国家:
    日本
  • 起止时间:
    2003 至 2004
  • 项目状态:
    已结题

项目摘要

Culturing of Spermatogonial Stem CellsIn April, 2003, it was reported that male germline stem cells, collected from neonatal mouse testes (strains of DBA/2 and ICR), were cultured for long period on feeder layer of mouse embryonic fibroblasts with 4 growth factors (EGF, bFGF, LIF, GDNF). We immediately repeated and confirmed results of the report and progressed it one step beyond by establishing germline stem cells (GS cells) from not only neonates but also from adult mice. The GS cells expanded exponentially in vitro. They produce sperm when transplanted recipient mouse testes. They did not produce tumor nor showed abnormal proliferation in the body of recipient mice. The GS cells was re-derived from recipient mouse testes, which showed that GS cells in vitro and spermatogonial stem cells in the testis were identical or mutually convertible.Fertilization of recipient mice with spermatogonial transplantationIn order to make recipient mice fertile after spermatogonial transplantation, immature mice (10-15 days old) were treated with irradiation before transplantation. The aim of irradiation was to make empty niche in the testis by removing endogenous germ cells for efficient colonization of donor stem cells. The dose of irradiation turned out to be optimal at around 12Gy for that purpose. One to Three days after the irradiation, the recipient mice received spermatogonial transplantation. Two months after the transplantation onward, the recipient mice were mated with females to test their fertility. It was confirmed that mice became fertile fathered pups of donor germ cell origin at high rate (about 70%). When we used GS cells, the fertility rate of recipient mice became higher, around 80%. The important factors for making recipient fertile with donor germ cells therefore include age of recipient, stem cell concentration of donor cell suspension.
精原干细胞的培养2003年4月,报道了从新生小鼠睾丸(DBA/2株和ICR株)采集的雄性生殖系干细胞,在含4种生长因子(EGF、bFGF、LIF、GDNF)的小鼠胚胎成纤维细胞饲养层上进行长期培养。我们立即重复和确认了报告的结果,并通过建立不仅来自新生儿的生殖系干细胞(GS细胞),也从成年小鼠建立了生殖系干细胞(GS细胞),使其更上一层楼。GS细胞在体外呈指数扩增。它们在移植受体小鼠的睾丸时会产生精子。它们在受体小鼠体内既没有产生肿瘤,也没有表现出异常增殖。从受体小鼠睾丸中分离出GS细胞,表明体外培养的GS细胞与睾丸内的精原干细胞是相同或可相互转化的。受体小鼠精原细胞移植的受精作用为了使受体小鼠在精原移植后生育,在移植前对未成熟小鼠(10-15天)进行照射。照射的目的是通过去除内源性生殖细胞在睾丸中留下空巢,以便有效地定植供体干细胞。为达到这一目的,照射剂量在12Gy时是最佳的。照射后1~3天,受体小鼠接受精原移植。移植两个月后,接受移植的小鼠与雌性交配,以测试它们的生育能力。已证实小鼠成为供体生殖细胞来源的可育父亲幼鼠的比率很高(约70%)。当我们使用GS细胞时,受体小鼠的受精率变得更高,约为80%。因此,受者年龄、供体细胞悬液中干细胞浓度是影响供体生殖细胞受精率的重要因素。

项目成果

期刊论文数量(28)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Increment of murine spermatogonial cell number by gonadotropin-releasing hormone analogue is independent of stem cell factor c-kit signal
  • DOI:
    10.1095/biolreprod.102.013276
  • 发表时间:
    2003-06-01
  • 期刊:
  • 影响因子:
    3.6
  • 作者:
    Ohmura, M;Ogawa, T;Sawada, H
  • 通讯作者:
    Sawada, H
Oligo-astheno-teratozoospemia in mice lacking Cnot7, a regulator of retinoid X receptor beta.
缺乏 Cnot7(视黄醇 X 受体 β 的调节剂)的小鼠出现少发性弱畸胎症。
  • DOI:
  • 发表时间:
    2004
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Nakamura T;Yao R;Ogawa T;et al.
  • 通讯作者:
    et al.
Derivation and morphological characterization of mouse spermatogonial stem cell lines
  • DOI:
    10.1679/aohc.67.297
  • 发表时间:
    2004-11-01
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Ogawa, T;Ohmura, M;Kubota, Y
  • 通讯作者:
    Kubota, Y
Identification and characterization of stem cells in prepubertal spermatogenesis in mice
  • DOI:
    10.1016/s0012-1606(03)00111-8
  • 发表时间:
    2003-06-01
  • 期刊:
  • 影响因子:
    2.7
  • 作者:
    Ohbo, K;Yoshida, S;Suda, T
  • 通讯作者:
    Suda, T
Abnormal sperm morphology caused by defects in sertoli cells of Cnot7 knockout mice
  • DOI:
    10.1679/aohc.67.307
  • 发表时间:
    2004-11-01
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Ogawa, T;Ito, C;Toshimori, K
  • 通讯作者:
    Toshimori, K
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OGAWA Takehiko其他文献

OGAWA Takehiko的其他文献

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{{ truncateString('OGAWA Takehiko', 18)}}的其他基金

In vitro human spermatogenesis using an organ culture method
使用器官培养方法进行体外人类精子发生
  • 批准号:
    24390371
  • 财政年份:
    2012
  • 资助金额:
    $ 1.34万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Solution of ill-posed inverse problems using complex-valuednetwork inversion
使用复值网络反演求解不适定反演问题
  • 批准号:
    21700260
  • 财政年份:
    2009
  • 资助金额:
    $ 1.34万
  • 项目类别:
    Grant-in-Aid for Young Scientists (B)
In vitro system for spermatogenesis from spermatogonial stem cells
精原干细胞产生精子的体外系统
  • 批准号:
    21592080
  • 财政年份:
    2009
  • 资助金额:
    $ 1.34万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Solution and regularization of inverse problems using complex-valued network inversion
使用复值网络反演的反演问题的求解和正则化
  • 批准号:
    18700231
  • 财政年份:
    2006
  • 资助金额:
    $ 1.34万
  • 项目类别:
    Grant-in-Aid for Young Scientists (B)
ex vivo spermatogenesis from cultured spermatogonial stem cells
培养精原干细胞的离体精子发生
  • 批准号:
    18591783
  • 财政年份:
    2006
  • 资助金额:
    $ 1.34万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Searching of factors facilitate spermatogonial stem cell proliferation (Investigation with transplantation and in vitro analysis)
寻找促进精原干细胞增殖的因素(移植研究和体外分析)
  • 批准号:
    13671663
  • 财政年份:
    2001
  • 资助金额:
    $ 1.34万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Analysis of spermatogonial proliferation with technique of spermatogonial transplantation-A trial for improving spermatogenic activity
精原细胞移植技术分析精原细胞增殖-提高生精活性的尝试
  • 批准号:
    11671569
  • 财政年份:
    1999
  • 资助金额:
    $ 1.34万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)

相似海外基金

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雄性生殖干细胞培养和基因改造
  • 批准号:
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  • 项目类别:
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  • 批准号:
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  • 财政年份:
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  • 批准号:
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  • 财政年份:
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  • 资助金额:
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  • 批准号:
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  • 财政年份:
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SPERMATOGONIAL STEM CELL CULTURE AND TRANSPLANTATION
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