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Method for testing that SLC can maintain homeostasis of Leydig and muscle-like cells in adult testis

A homeostasis and cell body technology, applied in the field of biochemical experiments, can solve problems such as unverified SLC

Pending Publication Date: 2020-12-22
THE SECOND HOSPITAL AFFILIATED TO WENZHOU MEDICAL COLLEGE
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  • Abstract
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  • Claims
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Problems solved by technology

But the hypothesis that SLCs may in fact be pluripotent, which can form Leydig or myoid lineages, depending on local regulatory factors including PDGF ligands, cannot currently be tested

Method used

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  • Method for testing that SLC can maintain homeostasis of Leydig and muscle-like cells in adult testis
  • Method for testing that SLC can maintain homeostasis of Leydig and muscle-like cells in adult testis
  • Method for testing that SLC can maintain homeostasis of Leydig and muscle-like cells in adult testis

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Embodiment Construction

[0052] This embodiment discloses a method for testing that SLC can maintain the homeostasis of Leydig and muscle-like myoid cells in adult testis, and the specific steps are as follows:

[0053] Step 1, the treatment of experimental animals: 90-day-old male SD rats were selected, the temperature of the SD rats was kept at 22°C, and light and dark were switched every 12 hours, and the SD rats were provided with free water. and food. Intraperitoneal injection of EDS (80mg / kg body weight) to the SD rats, after four days of EDS treatment, the EDS was prepared by dissolving 80mg / kg body weight in DMSO:PBS as a solvent of 1:3, by decapitation Kill SD rats;

[0054] Step 2. Isolation and culture of seminiferous tubules: put the testis tissue without albuginea in DMEM / F-12 medium, mechanically separate the seminiferous tubules from the interstitial tissue under a transmitted illumination dissecting microscope, and harvest them at equal lengths 5 cm isolated seminiferous tubules were...

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Abstract

The invention discloses a method for testing that SLC can maintain homeostasis of Leydig and muscle-like cells in adult testis. Stem cells related to testicular seminiferous tubules of adult SD rats are called as Leydig stem cells (SLC) and can be differentiated into Leydig cells in vitro and in vivo. Proliferation and differentiation of the SLC are regulated by locally generated factors, including PDGF family members PDGF-AA and PDGF-BB. Both members are able to stimulate proliferation of SLC, while only PDGF-AA is able to stimulate differentiation of SLC into Leydig cells producing testosterone (T), while PDGF-BB inhibits the process. In the method, it is verified that the SLC can actually form Leydig cells or myocyte-like cells under different differentiation conditions. PDGF-AA + LH can enable the SLC to be differentiated into Leydig cells capable of generating T, and PDGF-BB + TGFB induces the cells to become muscle-like cells for expressing smooth muscle actin. The method can effectively verify pluripotency of seminiferous tubule-associated stem cells (SLC), and test important role of SLC in maintaining homeostasis of Leydig and Myoid populations in vivo.

Description

technical field [0001] The invention relates to the technical field of biochemical experiments, in particular to a method for testing that SLC can maintain the homeostasis of Leydig and myoid cells in adult testes. Background technique [0002] The testes of mammals contain both germ cells and somatic cells. The germ cell population produces sperm, while the somatic cell population produces the male hormone testosterone (T) and supports sperm production. Leydig cells in the interstitium and myoid myoid cells around the seminiferous tubules are two important components of the testis. The main function of Leydig cells is to produce T, which is essential for maintaining sperm production in the testis and the male sex characteristics of the body. Defects in the development or maintenance of Leydig cells may lead to androgen deficiency (hypogonadism), thereby affecting the reproductive function of the individual as well as the physical and mental health of the individual. Hypo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02G01N33/68
CPCG01N33/502G01N33/5073G01N33/68
Inventor 陈浩林管小菊林函段萍吴越赵星星温馨季敏鹏陈丹赵星仪黄福
Owner THE SECOND HOSPITAL AFFILIATED TO WENZHOU MEDICAL COLLEGE
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