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Micro-needle based in vitro culture method for testicular tissue and applications thereof

A technology of in vitro culture and tissue culture, applied in tissue culture, culture process, cell culture active agent, etc., can solve the problem of inability to provide

Active Publication Date: 2021-08-31
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In order to solve the defects that the existing SSCs expansion method and the method for culturing testicular tissue in vitro cannot provide the required microenvironment for SSCs, the in vitro tissue culture center is prone to death, and the defects that the whole testis cannot be cultured, the present invention provides an in vitro cultured testicular tissue promoting A method for proliferating spermatogonial stem cells, said method being a non-therapeutic method, comprising the steps of:

Method used

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  • Micro-needle based in vitro culture method for testicular tissue and applications thereof
  • Micro-needle based in vitro culture method for testicular tissue and applications thereof
  • Micro-needle based in vitro culture method for testicular tissue and applications thereof

Examples

Experimental program
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Embodiment 1

[0062] Example 1: A method for culturing testicular tissue in vitro to promote the proliferation of spermatogonial stem cells

[0063] This embodiment provides a method for cultivating testicular tissue in vitro to promote the proliferation of spermatogonial stem cells. The method is not a therapeutic method, comprising the following steps:

[0064] Establishment of the culture system: soak the PEGDA microneedles in the culture medium for replacement to obtain a culture system containing the replaced PEGDA microneedles and the culture medium;

[0065] Culture of testicular tissue: add the isolated testicular tissue after removing the capsule to the culture system for cultivation, so that the spermatogonial stem cells in the isolated testicular tissue proliferate;

[0066] The PEGDA microneedle is a PEGDA microneedle prepared by using PEGDA with a volume fraction of 20%;

[0067] The soaking is: soaking at a temperature of 34°C for 24 hours;

[0068] The culture medium is a c...

Embodiment 2

[0073] Example 2: A method for culturing testicular tissue in vitro to promote the proliferation of spermatogonial stem cells

[0074] This embodiment provides a method for cultivating testicular tissue in vitro to promote the proliferation of spermatogonial stem cells. The method is not a therapeutic method, comprising the following steps:

[0075] Establishment of the culture system: soak the PEGDA microneedles in the culture medium for replacement to obtain a culture system containing the replaced PEGDA microneedles and the culture medium;

[0076] Culture of testicular tissue: add the isolated testicular tissue after removing the capsule to the culture system for cultivation, so that the spermatogonial stem cells in the isolated testicular tissue proliferate;

[0077] The PEGDA microneedle is a PEGDA microneedle prepared by using PEGDA with a volume fraction of 40%;

[0078] The soaking is: soaking for 12 hours at a temperature of 37°C;

[0079] The medium is SSC medium;...

Embodiment 3

[0084] Example 3: A method for culturing testicular tissue in vitro to promote the proliferation of spermatogonial stem cells

[0085] This embodiment provides a method for cultivating testicular tissue in vitro to promote the proliferation of spermatogonial stem cells. The method is not a therapeutic method, comprising the following steps:

[0086] Establishment of the culture system: soak the PEGDA microneedles in the culture medium for replacement to obtain a culture system containing the replaced PEGDA microneedles and the culture medium;

[0087] Culture of testicular tissue: add the isolated testicular tissue after removing the capsule to the culture system for cultivation, so that the spermatogonial stem cells in the isolated testicular tissue proliferate;

[0088] The PEGDA microneedle is a PEGDA microneedle prepared by using PEGDA with a volume fraction of 70%;

[0089] The soaking is: soaking at a temperature of 32°C for 20 hours;

[0090] The culture medium is a c...

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Abstract

The invention relates to a micro-needle based in vitro culture method for testicular tissue and applications thereof, and belongs to the technical field of biology. The method for in vitro culturing testicular tissue to promote proliferation of spermatogonial stem cells is provided. The method includes: soaking PEGDA micro-needles in a culture medium for replacement, so as to obtain a culture system including replaced PEGDA micro-needles and the culture medium; and adding isolated testicular tissue in the culture system to culture, so as to proliferate the spermatogonial stem cells in the isolated testicular tissue. The method improves situations of death of tissue centers during tissue culture in vitro, so that large tissue culture can be realized. Therefore, a physiological structure of the tissue can be maintained to the greatest extent, which is more in line with in vivo situations. In addition, the method can promote the proliferation of the spermatogonial stem cells in vitro, the self-renewal of the spermatogonial stem cells (SSCs) is supported, and the dryness of the spermatogonial stem cells can be maintained.

Description

technical field [0001] The invention relates to a microneedle-based method for culturing testicular tissue in vitro and its application, belonging to the field of biotechnology. Background technique [0002] Spermatogenesis originates from spermatogonial stem cells (SSCs). Proliferation and differentiation of spermatogonial stem cells is an orderly and continuous process, starting from puberty and continuing until death. Spermatogonial stem cells are adult tissue stem cells in the testes that underlie ongoing spermatogenesis and are critical for male fertility. SSCs mainly have the following two characteristics: [0003] 1) capable of self-renewal to maintain the stem cell pool; [0004] 2) Capable of differentiating to maintain the continuation of male / male spermatogenesis after puberty. [0005] Under normal circumstances, potential spermatogonial stem cells can rapidly divide into differentiated spermatogonia, and can be transformed into SSCs with self-renewal ability...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/076
CPCC12N5/061C12N2501/13C12N2501/115C12N2501/11C12N2501/16C12N2501/125C12N2501/155C12N2501/392C12N2501/31C12N2500/84C12N2501/998C12N2500/25C12N2500/46C12N2500/34C12N2500/38C12N2500/30C12N2500/44C12N2500/32
Inventor 沙家豪赵远锦袁艳郭雪江夏宇杨磊李来花朱云飞
Owner NANJING MEDICAL UNIV
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