Separation, purification and culture method of rat cornea endothelial cells

Abstract

The invention provides a separation, purification and culture method of rat cornea endothelial cells, and belongs to the field of cell biology. The method comprises the following steps: 1, separating a cornea tissue sample to obtain single cells; 2, preparing immunomagnetic beads for specifically adsorbing cornea endothelial cells, and purifying the cornea endothelial cells from the separated single cells; and 3, culturing the rat cornea endothelial cells with an optimized cell culture liquid. The separation process of the rat cornea endothelial cells has the advantages of simplicity in operation, high cell output and high cell survival rate, the purification process of the rat cornea endothelial cells has the advantages of simplicity in operation, very high yield and very high purification rate, and the culture process of the rat cornea endothelial cells has the advantages of fast cell proliferation, short period and high economic benefit.

Description

technical field [0001] The invention relates to a method for separating, purifying and culturing rat corneal endothelial cells, belonging to the field of cell biology. Background technique [0002] The morphological integrity and functional integrity of corneal endothelial cells are the indicators for clinical evaluation of corneal function and the standard for evaluating the quality of corneal preservation. Various corneal diseases, surgical injuries, and drug toxicity can cause damage to corneal endothelial cells. When the damage exceeds its physiological compensation, corneal lesions will occur, and corneal blindness will occur in severe cases. Therefore, observing the effects of drugs and other factors on the activity of corneal endothelial cells is inseparable from the in vitro experiments of corneal endothelial cells, and a good in vitro corneal endothelial cell isolation and culture method is the first step in in vitro experiments. The study of pharmacology, patholog...

AI Technical Summary

Problems solved by technology

[0004] At present, the in vitro primary culture of corneal endothelial cells includes: tissue block attachment method, enzyme digestion method (trypsin, collagenase and dispase enzyme), microdissection method, etc., but the operation of microdissection method is complicated; enzyme digestion method requires Strictly control enzyme digestion time and enzyme concentration to prevent enzyme damage to cells, while tissue adherence rules slow cell migration and growth rate

In the process of tissue isolation and cell culture, the problem of stromal cell contamination often occurs

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