Cell in-vitro culture method for primary culture of bovine mammary epithelial cells

Abstract

The invention discloses a cell in-vitro culture method for primary culture of bovine mammary epithelial cells.The cell in-vitro culture method comprises the following steps: pretreating bovine mammary tissues and then standing and carrying out adherent culture for 30 minutes by laterally placing a culture bottle; carrying out isolated culture by using a tissue block recycling method; and finally purifying by using a time-difference trypsin digestion method and a bottle internal digestion method to obtain the bovine mammary epithelial cells.The cell in-vitro culture method for primary culture of the bovine mammary epithelial cells has the beneficial effects that by the cell in-vitro culture method for primary culture of the bovine mammary epithelial cells, the purified bovine mammary epithelial cells can be obtained successfully, meanwhile, cell climbing-out time is shortened, cell proliferation speed is increased, cell growth state is good, possibility of cell pollution is greatly reduced, and the problem that beestings of mammary tissues during lactation period is not easy to clean and easy to pollute is avoided.

Description

technical field [0001] The invention relates to the technical field of cell engineering, in particular to an in vitro cell culture method for primary culture of dairy cow mammary gland epithelial cells. Background technique [0002] Mammary gland epithelial cells are the only cells with secretory function in the mammary gland. They are the basis for lactation and also an important part of the innate immune barrier of the mammary gland. Primary cells can well reflect their morphology, structure, and physiological and biochemical characteristics. The real situation in the body is suitable for research on physiology, pathology, and pharmacology. Breast epithelial cells have been used to study the mechanism of lactation, the pathogenesis of mastitis, and drug screening. An important tool for screening. With the wide application of tissue culture technology, the in vitro culture technology of dairy cow mammary gland epithelial cells has also been rapidly developed and applied. T...

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Problems solved by technology

The advantages of the tissue block culture method are low cost, simple operation, and strong growth and proliferation ability of the obtained cells; but the disadvantage is that it takes a long time and the subsequent purification work is difficult

The advantages of the mechanical disruption method are simple operation, low cost, high cell yield and rapid growth; the disadvantage is that there is some mechanical damage to the cells and subsequent purification is required

The advantage of the enzymatic digestion method is that the cells proliferate faster and the purity is higher; the disadvantage is that the enzyme will cause certain damage to the cells, so the concentration of the enzyme and the digestion time must be strictly controlled during the operation

The advantage of the milk extraction method is that the obtained cells have high purity, good activity, and a short time. However, there are still some problems in this method, such as the need to correctly grasp the sampling time, high requirements on the quality and quantity of milk, and the number of cells obtained. Less, there will be some immune cell contamination, etc.

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