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Method for inducing pluripotent stem cells by bovine sertoli cells

A technology of Sertoli cells and pluripotent stem cells, applied in the field of stem cells, can solve the problems of incomplete pluripotency and low induction efficiency, and achieve the effect of easy cell acquisition and high reprogramming rate

Inactive Publication Date: 2019-05-03
JILIN UNIV
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Problems solved by technology

[0005] In view of the deficiencies in the prior art, the object of the present invention is to provide a method for inducing pluripotent stem cells from bovine testicular Sertoli cells, which solves the problems of low induction efficiency of bovine induced pluripotent stem cells and incomplete pluripotency of cells, etc.

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  • Method for inducing pluripotent stem cells by bovine sertoli cells
  • Method for inducing pluripotent stem cells by bovine sertoli cells
  • Method for inducing pluripotent stem cells by bovine sertoli cells

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Embodiment 1

[0027] Preparation of Sertoli cells from bovine testis:

[0028] The calf testes retrieved from the slaughterhouse were used for the primary isolation of Sertoli cells. The specific operation was as follows: the seminiferous tubule seminiferous epithelial cell suspension of bovine testis was prepared by two-step enzyme digestion method: under sterile conditions, the The meat calf testicles retrieved from the slaughterhouse were fully stripped, and 5 times the volume of DMEM medium was added and blown vigorously to form a single seminiferous tubule as much as possible. After standing for 5 minutes, the supernatant was discarded, and repeated 5 times to remove the testes. Add 5 times the volume of digestion solution I containing 50 μg / mL deoxyribonuclease I (DNAse I) and 1 mg / mL collagenase IV after the upper layer of liquid is clarified, and incubate on a constant temperature shaker at 32°C for 60 min at a speed of 140 rpm; The DMEM medium containing 10% fetal bovine serum was ...

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Abstract

The invention discloses a method for inducing pluripotent stem cells by bovine sertoli cells, and belongs to the field of stem cells. The method comprises the following steps: step 1, separating bovine sertoli cells; step 2, cultivating the bovine sertoli cells; and step 3, transferring four transcription factors encoding Oct4, Sox2, c-Myc and Klf4 into the bovine sertoli cells so as to produce the pluripotent stem cells. Compared with the prior art, the method fully utilizes the characteristics of differential adherence of the bovine sertoli cells, and cultivates the bovine sertoli cells withuniform morphology. Since the bovine sertoli cells are derived from meat calf, the bovine sertoli cells have a high reprogramming ability. The bovine sertoli cells replace bovine adult fibroblasts and mesenchymal stem cells, and are used as source cells for preparing and inducing pluripotent stem cells, so that not only the cell acquisition is simple, but also the reprogramming rate is high.

Description

technical field [0001] The invention belongs to the field of stem cells, and in particular relates to a method for inducing pluripotent stem cells from bovine testicular Sertoli cells. Background technique [0002] Embryonic stem cells (Embryonic Stem Cells, ESCs) are isolated from the inner cell mass (Inner cell mass, ICM), a kind of pluripotent stem cells that can differentiate into various types of cells of the three germ layers. Significant progress has been made in the establishment, in vitro culture, and molecular studies of mouse, rat, and human ESCs. However, for large livestock, especially bovine ESCs, researchers have tried a variety of in vitro isolation and culture methods on the basis of mouse ESCs. However, due to the large differences between species, the composition of in vitro medium is uncertain, and the source of egg cells is inconvenient. True bovine ESCs have not been reported yet. Since the cloned sheep Dolly was born on July 5, 1996, somatic cell nuc...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074C12N5/071
Inventor 张学明姜禹李子义安星兰蔡宁宁张胜赵欣欣朱文倩唐博
Owner JILIN UNIV
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