Method for establishing lactation model of cow mammary gland epithelial cells
A technology for mammary epithelial cells and cells, which is applied in the field of biological cytology, can solve the problems of immature in vitro culture technology of dairy cow mammary epithelial cells, and achieve the effects of low culture cost, low culture condition requirements and high survival rate.
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Embodiment 1
[0024] Embodiment 1: a kind of method of the present invention establishes cow mammary gland epithelial cell lactation model, the experimental method step is as follows:
[0025] Step 1: Primary culture and identification of dairy cow mammary gland epithelial cells
[0026] Disinfect the middle part of the breast, cut out a small amount of breast tissue according to conventional surgery, immediately wash the tissue with D-Hanks solution and peel off the fat tissue and connective tissue as much as possible, and cut the breast acinar tissue into 1mm 3 Inoculate small pieces of collagen in the cell culture flask pre-coated with collagen at a distance of 0.5 cm. After covering the entire bottle, culture it upside down at 37°C 2 In the incubator for about 3-4 hours, until the tissue block is firmly attached to the collagen, gently add the growth medium to the culture, so as to cover the tissue block, then place it in the incubator to continue the culture, and replace the culture ev...
Embodiment 2
[0033] Example 2: Combining Figure 1-Figure 6 , the experiment done by the present invention:
[0034] Experimental material: healthy Holstein cow mammary gland tissue. The breast tissues sampled were in puberty for 2 months, lactation period for 7 days, lactation period for 280 days, dry period for 3 days, and perinatal period for 10 days. Experimental reagents: rat tail collagen, Hangzhou Shengyou; high-quality fetal bovine serum, DF12 medium, Gibco; cytokeratin 18 antibody, Acris, Germany; PI dye, insulin, Sigma; chromatography grade methanol, Tianjin Guangfu; lactose standard Products, Academy of Military Medical Sciences; β-casein standard, Sigma. Experimental equipment: ZHJH-1112 vertical flow ultra-clean bench, Shanghai Zhicheng Analytical Instrument Manufacturing Co., Ltd.; CO-150 CO 2 Incubator, Japan; SZ-93 automatic double pure water distiller, Shanghai Yarong Biochemical Instrument Factory; LC-10AT high performance liquid chromatography, Shimadzu, Japan, SPD-10...
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