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Vector capable of promoting self renewal and proliferation of spermatogonial stem cells of dairy goat and application of vector

A technology of spermatogonial stem cells and cell proliferation, which is applied in the field of self-renewal and proliferation carriers, can solve the problems of adult tissue stem cells, etc., and achieve the effect of increasing self-renewal ability, accelerating cell proliferation, and promoting self-renewal and proliferation

Inactive Publication Date: 2015-03-11
NORTHWEST A & F UNIV
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Problems solved by technology

Although the in vitro culture and expansion of pluripotent ES cells has become increasingly routine, the establishment of in vitro culture techniques for adult tissue stem cells (including mGSCs) is still difficult

Method used

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  • Vector capable of promoting self renewal and proliferation of spermatogonial stem cells of dairy goat and application of vector
  • Vector capable of promoting self renewal and proliferation of spermatogonial stem cells of dairy goat and application of vector
  • Vector capable of promoting self renewal and proliferation of spermatogonial stem cells of dairy goat and application of vector

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Embodiment Construction

[0032] The present invention will be further described in detail below in conjunction with specific embodiments, which are explanations of the present invention rather than limitations.

[0033] The carrier for promoting the self-renewal and proliferation of dairy goat spermatogonial stem cells provided by the present invention uses the Sirt1 gene related to spermatogenesis as the target gene to construct a eukaryotic expression vector, and uses dairy goat spermatogonial stem cells as a host for transduction to promote spermatogenesis. Overexpression or overexpression of the gSirt1 gene in primary stem cells.

[0034] Specifically, the present invention first constructs the eukaryotic expression vector pSirt1-IRES2-AcGFP containing gSirt1 and green fluorescent protein (AcGFP) genes, transfects the vector into male reproductive stem cells of dairy goats, and observes marker gene AcGFP and Sirt1 antibody staining under a fluorescent microscope Its expression was detected, and it...

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Abstract

The invention discloses a vector capable of promoting the self renewal and proliferation of spermatogonial stem cells of a dairy goat and application of the vector. The vector is an eukaryotic expression vector containing a dairy goat gSirt1 gene, wherein the nucleotide sequence of the dairy goat gSirt1 gene is shown as SEQ.ID.NO.1. According to the vector and the application of the vector, over-expression or super expression in the spermatogonial stem cells is realized by the constructed eukaryotic expression vector, and RT-PCR (Reverse Transcription-Polymerase Chain Reaction), Western blot and cell cycle examination show that the gSirt1 gene is helpful for the self renewal, stemness (pluripotency) maintenance and cell proliferation of the spermatogonial stem cells.

Description

technical field [0001] The invention belongs to the technical field of spermatogonial stem cells, and relates to a carrier for promoting self-renewal and proliferation of dairy goat spermatogonial stem cells and an application thereof. Background technique [0002] Spermatogonial stem cells (SSCs), also known as male germline stem cells (mGSCs), are a type of adult stem cells located on the basement membrane of the seminiferous tubules of the testis. SSCs can maintain their own population through self-renewal The number is stable, and can develop and differentiate to form sperm cells. Studies have shown that in the microenvironment of the testis, SSCs, like embryonic stem cells, have the potential to differentiate into all cell types including the three germ layers in vitro and in vivo (Oatley JM, Brinster RL. Regulation of spermatogonial stem cell self-renewal in mammals. Annu Rev Cell Dev Biol 2008;24:263-86.). Spermatogonia stem cells can be used as an ideal in vitro mo...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N5/10
Inventor 华进联吴江牛博文
Owner NORTHWEST A & F UNIV
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