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Method for building dairy cattle breast acinus lactation model in vitro

A construction method and mammary gland technology, applied in the field of in vitro construction of lactation models, can solve the problems of low authenticity of simulated in vivo environment, and achieve the effect of increasing intuition and high authenticity

Inactive Publication Date: 2010-10-13
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to solve the problem of low authenticity of the simulated in vivo environment with mammary epithelial cells as the research carrier when studying the lactating function of the dairy cow mammary gland in vitro, and to provide an in vitro construction method of the milk cow mammary gland acinar lactation model

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  • Method for building dairy cattle breast acinus lactation model in vitro
  • Method for building dairy cattle breast acinus lactation model in vitro
  • Method for building dairy cattle breast acinus lactation model in vitro

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specific Embodiment approach 1

[0013] Specific embodiment one: the in vitro construction method of dairy cow mammary gland acinar lactation model according to the following steps in this embodiment: one, matrix (Growthfactor-reduced Matrigel TM ) was melted at 4°C, and spread on a pre-cooled cell culture plate at 4°C, the matrix thickness of each well was 0.5 mm, and then placed in a cell culture incubator at 37°C for 30-60 minutes to obtain a matrix gel cell culture plate ; 2. Using trypsin with a mass concentration of 0.25% and EDTA with a mass concentration of 0.02% to digest primary cultured milk cow mammary gland epithelial cells at 4 months of pregnancy for 5 minutes, and then centrifuged at 1500r / min for 5 minutes to collect the cells; 3. The collected cells were suspended in 4ml of assay medium to prepare a cell suspension, and then diluted with assay medium to a density of 1×10 4 ; 4. Add 2ml of matrix gel to the cell culture plate with a density of 1×10 4 The cell suspension was placed in an incu...

specific Embodiment approach 2

[0027] Embodiment 2: This embodiment differs from Embodiment 1 in that in step 1, place it in a cell culture incubator at 37° C. for 40 to 50 minutes to obtain a matrix gel cell culture plate. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0028] Embodiment 3: This embodiment differs from Embodiment 1 in that in Step 1, place it in a cell culture incubator at 37° C. for 45 minutes to obtain a matrix gel cell culture plate. Other steps and parameters are the same as those in Embodiment 1.

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Abstract

The invention discloses a method for building a dairy cattle breast acinus lactation model in vitro, and relates to a method for building a lactation model in vitro. The method solves the problem of low authenticity of in-vivo environment caused by taking a mammary epithelial cell as a research carrier in the existing in-vitro study of the lactation function of the dairy cattle breast. The method comprises the following steps: 1, preparing a tissue culture plate of matrix gel; 2, digesting primary cultured dairy cattle mammary epithelial cell gestated for 4 months by adopting pancreatin and EDTA, and collecting cells after centrifugalization; 3, preparing cell suspension and diluting; and 4, adding the cell suspension into the tissue culture plate of the matrix gel, culturing for 15 days in an incubator to finish the building. The artificial building process of the dairy cattle breast acinus lactation model built by the invention is a simulation process of in-vitro simulation of a galactemia protective screen; and a three-dimensional acinus structure thereof can truly simulate in-vivo situation, has the same lactation function and biological characteristics as the breast acinus in a ruminant, and has high authenticity of in-vivo environment simulation.

Description

technical field [0001] The invention relates to a method for constructing a lactation model in vitro. Background technique [0002] The mammary gland is where milk is synthesized and secreted. In the past, mammary gland epithelial cells cultured on a two-dimensional platform were used as the research carrier to study the function and regulation of mammary gland lactation in vitro. However, studies have shown that once mammary epithelial cells are isolated and placed in two-dimensional (planar) conditions for culture, they will soon lose their tissue-specific functions. Although prolactin can activate STAT5 (signal transducer and activator of transcription 5) in the two-dimensional culture of mammary epithelial cells, the activation of STAT5 is transient and cannot induce the expression of milk protein genes. However, the mammary acinus, as the smallest functional unit of lactation, is different. The acinar structure can not only express prolactin receptors on the basal sid...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 李庆章林叶侯晓明高学军
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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