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Livestock spermatogonial stem cell medium

A spermatogonial stem cell and culture medium technology, applied to the field of livestock spermatogonial stem cell culture medium, can solve the problems of low success rate and the like

Active Publication Date: 2013-05-01
INNER MONGOLIA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Not to mention its low success rate, the impact of epigenetic changes in the process of animal cloning on the genetic phenotype of offspring has not yet been resolved

Method used

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  • Livestock spermatogonial stem cell medium
  • Livestock spermatogonial stem cell medium
  • Livestock spermatogonial stem cell medium

Examples

Experimental program
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Effect test

Embodiment 1

[0050] Embodiment 1, livestock spermatogonia stem cell culture medium (SSCM)

[0051] A. Components of SSCM

[0052] DMEM / F12 (high glucose) powder, sodium pyruvate, L-glutamin, BSA, Biotin, DL AlphaTocopherol Acetate, DL Alpha-Tocopherol (vit.E), catalase, human recombinant insulin, human transferrin, peroxidase Oxygen dismutase, corticosterone (corticosteroid), galactose, ethanolamine / ethanolamine-C1, glutathione, L-carnitine, linoleic acid, linolenic acid, progesterone, putrescine / putrescine-2HCl, Sodium selenite, triiodothyronine, selenomethionine, GDNF, bFGF, Gfral, MEM NON-ESSENTIAL AMINO ACIDS SOLUTION(100X), BME AMINO ACIDS SOLUTION(50X), penicillin stock solution, streptomycin stock solution, Fungizone stock solution.

[0053] The preparation method of the basal culture medium SSCB of B.SSCM

[0054] B1. Prepare 1 liter of SSCB solution according to the formula in the table below, and strictly follow the technical requirements for aseptic operation of cell culture....

Embodiment 2

[0067] Embodiment 2 Application of Livestock Spermatogonia Stem Cell Culture Fluid

[0068] A. Preparation steps of trophoblast cells

[0069] 1. STO cells were cultured with STO cell culture medium (DMEM / F12, supplemented with 7% standard fetal bovine serum, penicillin 100 U / ml, streptomycin 100 μg / ml). Passaging at a ratio of 1:4 each time, the confluence of the cells reaches 70%-80% in about 2 days, ready for mitomycin treatment.

[0070] 2. Cultivate the STO cells to a confluence of about 80%, add mitomycin (calculated at a final concentration of 15 μg / ml), put them in a cell culture incubator at 37°C, 5% CO 2 , cultivated for 2 h under the condition of saturated humidity.

[0071] 3. Remove the culture solution containing mitomycin, and wash with 10ml PBS three times.

[0072] 4. Use 1ml Trypsin / EDTA to detach the cells, and use 4ml culture solution D to stop the digestion of Trypsin, collect the digested cells, and centrifuge at 2000rpm for 5 minutes.

[0073] 5. Res...

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Abstract

The invention discloses a livestock spermatogonial stem cell medium (SSCM). The preparation method of the livestock SSCM comprises a step of uniformly mixing 1ml of SSCB, 1mul of each of three cytokine stock solutions, 1mul of a penicillin stock solution, 1mul of a streptomycin stock solution and 1mul of a Fungizone stock solution before use. The livestock SSCM is suitable for the long-term culture of livestock SSCs. The invention simultaneously discloses a livestock SSC long-term culture method, a livestock SSC immunofluorescence dyeing identification method, a livestock SSC cryopreservation method and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a culture solution of livestock spermatogonial stem cells. Background technique [0002] Spermatogonia stem cells (SSCs) are one of the most important adult stem cells. Spermatogonial stem cells not only have the genetic function of procreation, recent studies have shown that spermatogonial stem cells can easily produce pluripotent stem cells through dedifferentiation. This makes spermatogonial stem cell manipulation technology favored by experts in life sciences, agricultural sciences and medicine [1]. Since the establishment of spermatogonial stem cell transplantation technology [2,3], research on spermatogonial stem cells has been unprecedentedly developed. In recent years, long-term in vitro culture methods of mouse and rat spermatogonial stem cells have been established [4,5]. To operate spermatogonial stem cells in vitro, the spermatogonial stem cells must first be isolated ...

Claims

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Application Information

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IPC IPC(8): C12N5/076
Inventor 吴应积罗奋华张岩萨初拉苏慧敏刘陶迪于泊洋刘林洪包佳婧孔群芳刘代艳刘燕
Owner INNER MONGOLIA UNIVERSITY
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