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Culture medium for accelerating in vitro differentiation of mammary epithelial cells of dairy cattle and application thereof

A breast epithelial cell and breast epithelial technology, applied in the field of cell biology, can solve the problems of low expression level and ineffective effect

Inactive Publication Date: 2013-06-12
INNER MONGOLIA AGRICULTURAL UNIVERSITY +1
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Problems solved by technology

[0002] With the development of biological technology, cow mammary epithelial cells cultured in vitro can be used as an ideal model to study the expression of milk protein genes. The problem is that most researchers currently use DMEM / F12 medium when cultivating dairy cow mammary epithelial cells. Under the condition of using DMEM / F12 medium, the expression levels of CSN1S1 and CSN3 genes in dairy cow mammary epithelial cells are low, indicating that this medium promotes In vitro differentiation and culture of dairy cow mammary gland epithelial cells is not obvious

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  • Culture medium for accelerating in vitro differentiation of mammary epithelial cells of dairy cattle and application thereof
  • Culture medium for accelerating in vitro differentiation of mammary epithelial cells of dairy cattle and application thereof
  • Culture medium for accelerating in vitro differentiation of mammary epithelial cells of dairy cattle and application thereof

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Embodiment Construction

[0016] 1.1 Main reagents

[0017] Fetal bovine serum (Gibco10099-141), 0.25% Trypsin-EDTA (Gibco25200-056), DMSO (Sigma D2650), double antibody (Gibco15140-122), insulin transferrin (Gibco51500-056), collagenase II (Gibco17101- 015), EGF (Gibco PHG-0313), DPBS (Hyclone SH30028-01B), MTT (AMRES10), prolactin (Sigma L6520), hydrocortisone (Sigma H0135), L-glutamine (Sigma G-8540 ), keratin 18 mouse monoclonal antibody, goat anti-mouse IgG-FITC (Shanghai Sangong), SYBR Premix Ex Taq TM II (TaKaRa DRR081), total cellular RNA extraction kit (TIANGEN DP430).

[0018] 1.2 Culture of primary mammary epithelial cells

[0019] Select healthy Chinese Holstein dairy cows in the mid-lactation stage, collect appropriate amount of mammary gland deep tissue (pay attention to avoid connective tissue and adipose tissue), put it in DMEM / F12 medium with double antibody, put it in an incubator and bring it back to the laboratory quickly . Rinse the mammary gland tissue block 3 times with DPBS s...

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Abstract

The invention discloses a culture medium for accelerating in vitro differentiation of mammary epithelial cells of dairy cattle and application thereof. Based on a DMEM (Dulbeccos Modified Eagles Medium) / F12 formula released by the Invitrogen company, composition of amino acids in the culture medium can be improved. The activity of the culture medium to mammary epithelial cells is remarkably higher than that of a DMEM / F12 culture medium. Genetic expression of mammary epithelial cells CSN1S1 and CSN3 is remarkably up-regulated (P is less than 0.05) compared with that of the commercial DMEM / F12 culture medium. The up-regulated amounts respectively reach 3.57 and 3.27 times. In addition, the culture medium provided by the invention is free from negative effect to proliferation of mammary epithelial cells of dairy cattle, lactation related genetic expression and casein synthesis.

Description

technical field [0001] The invention belongs to the field of cell biology, and in particular relates to a culture medium for promoting the differentiation of dairy cow mammary gland epithelial cells in vitro and application thereof. Background technique [0002] With the development of biological technology, cow mammary epithelial cells cultured in vitro can be used as an ideal model to study the expression of milk protein genes. The problem is that most researchers currently use DMEM / F12 medium when cultivating dairy cow mammary epithelial cells. Under the condition of using DMEM / F12 medium, the expression levels of CSN1S1 and CSN3 genes in dairy cow mammary epithelial cells are low, indicating that this medium promotes The effect of in vitro differentiation culture of dairy cow mammary epithelial cells was not obvious. According to the DMEM / F12 formula published by Invitrogen, the present invention improves the culture medium, thereby achieving the purpose of promoting th...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 张兴夫
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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