Method for differentiating bone cells by oriented inducing chick embryo stem spermatogonium

A spermatogonial stem cell and osteoblast technology, which is applied in the field of directional induction of chicken embryo spermatogonial stem cells to differentiate into osteoblasts, and achieves the effect of strong operability, advanced and simple technology

Inactive Publication Date: 2007-03-28
YANGZHOU UNIV
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Problems solved by technology

[0004] However, the research on poultry SSCs is still in its infancy. The research on chicken embryo SSCs is more common in isolation methods, extraction and purification, obtaining chicken spermatogonial stem cells, in vitro culture of various cytokines, cryopreservation of SSCs, and transplantation of SSCs or testicular tissues. However, there is no report on its ability to induce differentiation into various cells in vitro

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  • Method for differentiating bone cells by oriented inducing chick embryo stem spermatogonium

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Embodiment Construction

[0011] Step 1: Isolation and purification of chicken embryo SSCs

[0012] Take chicken embryos hatched for 16-19 days, and obtain chicken testes aseptically. Digested with collagenase (1 mg / ml) and then digested with trypsin (0.25%) to obtain SSCs, Sertoli cells and mesenchymal cells on the seminiferous epithelium, and then filtered through a 350-mesh filter, percoll gradient centrifugation, and paste Differential wall velocity to obtain higher purity spermatogonial stem cells, stain with trypan blue and count, adjust the cell density to 1x10 5 cells / ml, inoculated in 6-well culture plate, placed at 38°C, 5% CO 2 cultured in an incubator.

[0013] Step 2: Induction culture of chick embryonic SSCs in vitro

[0014] After SSCs were cultured for 3-4 days, preheated conditional induction medium (basal medium+10 -7 mol / L dexamethasone + 0.01mol / L β-sodium glycerophosphate + 0.05g / L vitamin C) for half-quantity fluid change. After that, observe every 24 hours, and change the li...

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Abstract

The invention discloses the method used to directionally induce chicken embryo stem spermatogonium to differentiate into bone cell. Its features are that inoculating the chicken stem spermatogonium to culture bottle without feeder layer; using conditioned medium to induce culturing which is that the DMEM high glucose contains 10% calf blood serum, 2% chicken blood serum, 2mmol/L L-glutamine, 1mmol/L sodium pyruvate, 5.5*10-15ng/ml beta-mercaptoethanol, 10 mul/ml non-essential amino acid, 10-7mol/L dexamethasone, 0.01mol/L beta-phospho glycerol, 0.05g/L vitamin C. The invention has the advantages of advanced technology, simple operation, strong repeatability, providing possible for realizing tissue organ rebirthing and replacing. Thereby hopefully to find out the approach for build cell platform in large scale production, further a new study prospect can be exploited in developmental biology, medicine, genetics fields etc.

Description

technical field [0001] The invention relates to a method for differentiating chicken embryo spermatogonial stem cells into osteoblasts, which belongs to cell engineering [0002] technology field. Background technique [0003] Chicken spermatogonial stem cells (SSCs) have the ability to differentiate into pluripotency, and adding appropriate differentiation-inducing agents during in vitro culture can induce the required special types of cells and tissues, which has attracted great attention. At present, people's research on the induction of stem cells into osteoblasts is mostly limited to mammals (rats, mice). Gopalakrish.V et al. used mouse bone marrow stromal stem cells (BMSCs) to induce osteoblasts. Study the internal mechanism of BMSCs induction of osteoblasts; Zhang Weicheng, Ding Lianghua, etc. respectively used rat and human bone marrow stromal cells to induce osteoblasts, and they all achieved success. [0004] However, the research on poultry SSCs is still in its ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/077
Inventor 李碧春孙国波陈国宏吴信生赵文明徐琪葛剑辉魏彩霞戴建明孙鹏翔
Owner YANGZHOU UNIV
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