Male germline stem cell culture and genetic modification
雄性生殖干细胞培养和基因改造
基本信息
- 批准号:7228882
- 负责人:
- 金额:$ 33.81万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2003
- 资助国家:美国
- 起止时间:2003-07-01 至 2009-04-30
- 项目状态:已结题
- 来源:
- 关键词:ActinsAdultAffectAnimalsBiochemicalBiological AssayBiologyCell CycleCell LineCell SeparationCell TransplantationCellsCellular biologyChickensDependovirusDevelopmentDoxycyclineElongation FactorFluorescence-Activated Cell SortingGalactosidaseGene MutationGenesGeneticGenetic ProgrammingGerm CellsGlycoproteinsGreen Fluorescent ProteinsIn VitroLacZ GenesMagnetismMethodsModificationMolecularMoloney Leukemia VirusMusPhosphoglycerate KinasePopulationProcessReporterScientistSeminiferous tubule structureSeriesSpermatogenesisSpermatogoniaStagingStem cellsSystemTechniquesTestisTetanus Helper PeptideTetracyclineTetracyclinesTimeTransplantationVesicular stomatitis Indiana virusVirusanalogcell typeimmortalized cellmalemature animalpromoterpupresearch studysertoli cell
项目摘要
DESCRIPTION (provided by applicant): Development of the testis cell transplantation technique established a system to study the biology of spermatogonial stem cells (SSCs) as well as the differentiation processes that occur during spermatogenesis. Currently there are no biochemical, molecular or morphological criteria by which the SSC can be identified, and only a functional assay can establish the presence of the SSC in any cell population. During the past eight years the transplantation technique has demonstrated that the stem cell can be cryopreserved, transplanted from many species, maintained in vitro, and subjected to genetic modification. The technique also has been used to establish the cell type (germ cell or Sertoli cell) responsible for both natural and induced genetic mutations, and to demonstrate that cell cycle timing during spermatogenesis is controlled by the genetic program of the germ cell. Thus, the transplantation system has provided a powerful approach to study the stem cell, its niche in the seminiferous tubule, and the process of spermatogenesis.
Despite these advances, it is not possible to obtain and study pure populations of SSCs from testes of mature animals, which greatly impedes studies on the biology of these cells. We propose a series of experiments in the mouse to further enrich testis cell populations for stem cell content and then to use these purified populations for introduction of genetic modifications into the SSCs. An important group of genes we plan to examine for their effect are those that have been shown to immortalize stem cells. This approach will provide us with two populations of germ cells: 1) testis cell populations isolated directly from the animal and greatly enriched for stem cells, and 2) immortalized cell lines that can be propagated in vitro as stem cells or early stage spermatogonia. The ability of these cells to colonize the seminiferous tubules of recipient animals and generate spermatogenesis as well as their ability to differentiate in vitro will be used to characterize the original cell populations and to study the differentiation process. The transplantation technique has been a productive approach in our studies over the past eight years and has proven increasingly valuable to other scientists. The studies outlined will continue progress toward our objective of understanding the biology of SSCs and spermatogenesis, and many of our findings will be applicable to other species.
描述(由申请人提供):睾丸细胞移植技术的发展建立了一个系统来研究精原干细胞(SSC)的生物学以及精子发生过程中发生的分化过程。目前,尚无可鉴定 SSC 的生化、分子或形态学标准,只有功能测定才能确定任何细胞群中是否存在 SSC。在过去的八年中,移植技术已经证明干细胞可以冷冻保存、从许多物种移植、在体外保存并进行基因改造。该技术还被用来建立负责自然和诱导基因突变的细胞类型(生殖细胞或支持细胞),并证明精子发生过程中的细胞周期时间是由生殖细胞的遗传程序控制的。因此,移植系统为研究干细胞、其在生精小管中的生态位以及精子发生过程提供了强有力的方法。
尽管取得了这些进展,但不可能从成熟动物的睾丸中获得并研究纯的 SSC 群体,这极大地阻碍了对这些细胞的生物学研究。我们建议在小鼠中进行一系列实验,以进一步富集睾丸细胞群的干细胞含量,然后使用这些纯化的细胞群将遗传修饰引入 SSC。我们计划检查其作用的一组重要基因是那些已被证明可以使干细胞永生的基因。这种方法将为我们提供两种生殖细胞群:1)直接从动物中分离出的睾丸细胞群,并且富含干细胞;2)可以作为干细胞或早期精原细胞在体外繁殖的永生化细胞系。这些细胞定殖于受体动物的生精小管并产生精子发生的能力以及它们在体外分化的能力将用于表征原始细胞群并研究分化过程。在过去的八年里,移植技术在我们的研究中一直是一种富有成效的方法,并且事实证明对其他科学家来说越来越有价值。概述的研究将继续朝着我们了解 SSC 生物学和精子发生的目标取得进展,并且我们的许多发现将适用于其他物种。
项目成果
期刊论文数量(15)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Ralph Lawrence Brinster其他文献
Ralph Lawrence Brinster的其他文献
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{{ truncateString('Ralph Lawrence Brinster', 18)}}的其他基金
microRNA regulation of spermatogonial stem cell self-renewal and differentiation
microRNA对精原干细胞自我更新和分化的调控
- 批准号:
8532010 - 财政年份:2012
- 资助金额:
$ 33.81万 - 项目类别:
microRNA regulation of spermatogonial stem cell self-renewal and differentiation
microRNA对精原干细胞自我更新和分化的调控
- 批准号:
8214793 - 财政年份:2012
- 资助金额:
$ 33.81万 - 项目类别:
Regulation of mouse spermatogonial stem cell self-renewal
小鼠精原干细胞自我更新的调控
- 批准号:
7933520 - 财政年份:2009
- 资助金额:
$ 33.81万 - 项目类别:
Regulation of mouse spermatogonial stem cell self-renewal
小鼠精原干细胞自我更新的调控
- 批准号:
7647221 - 财政年份:2007
- 资助金额:
$ 33.81万 - 项目类别:
Regulation of mouse spermatogonial stem cell self-renewal
小鼠精原干细胞自我更新的调控
- 批准号:
8090465 - 财政年份:2007
- 资助金额:
$ 33.81万 - 项目类别:
Regulation of mouse spermatogonial stem cell self-renewal
小鼠精原干细胞自我更新的调控
- 批准号:
7305510 - 财政年份:2007
- 资助金额:
$ 33.81万 - 项目类别:
Regulation of mouse spermatogonial stem cell self-renewal
小鼠精原干细胞自我更新的调控
- 批准号:
7485603 - 财政年份:2007
- 资助金额:
$ 33.81万 - 项目类别:
Regulation of mouse spermatogonial stem cell self-renewal
小鼠精原干细胞自我更新的调控
- 批准号:
7874705 - 财政年份:2007
- 资助金额:
$ 33.81万 - 项目类别:
Male germline stem cell culture and genetic modification
雄性生殖干细胞培养和基因改造
- 批准号:
6873662 - 财政年份:2003
- 资助金额:
$ 33.81万 - 项目类别:
Male germline stem cell culture and genetic modification
雄性生殖干细胞培养和基因改造
- 批准号:
6755916 - 财政年份:2003
- 资助金额:
$ 33.81万 - 项目类别:
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