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Human renal tubular epithelial cell separation and culture method

A technology of epithelial cells and culture methods, applied in the field of cell biology, can solve problems affecting experimental results, cell variation, and long time of cell strains in vitro, so as to simplify operation steps, improve stability, reliability, and repeatability strong effect

Inactive Publication Date: 2019-03-01
JIANGYIN CHI SCI
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  • Abstract
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  • Claims
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Problems solved by technology

At present, the research on renal tubular epithelial cells mostly adopts cell lines with low cultivation difficulty and short preparation time, but the cell lines take a long time in vitro, the cells will mutate and affect the experimental results, and the primary culture has the advantage of being closer to the state in vivo

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  • Human renal tubular epithelial cell separation and culture method

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Embodiment Construction

[0009] In order to demonstrate the purpose and advantages of the present invention more clearly, specific embodiments are now further described. The specific embodiments described here are only for explaining the present invention, and are not intended to limit the present invention.

[0010] The invention selects fresh human kidney tissue and separates human renal tubular epithelial cells. The specific operation is as follows:

[0011] 1. Take out the kidney tissue under aseptic conditions, place it in a petri dish containing pre-cooled saline and double antibodies, wash it repeatedly with sterile PBS for 3-4 times, and separate the renal cortex and renal medulla;

[0012] 2. Use sterile ophthalmic scissors to cut the renal cortex to about 1mm 3 The small pieces were washed 2-3 times with sterile PBS, centrifuged at 1500 rpm / min for 5 min, and the supernatant was discarded;

[0013] 3. Place the tissue block on overlapping stainless steel sieves with apertures of 80 mesh a...

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Abstract

The invention provides a human renal tubular epithelial cell separation and culture method which comprises the following steps: (a) cleaning a tissue specimen with a precooling PBS (phosphate buffer solution) containing double antibodies, (b) shearing and grinding a tissue, and collecting ground tissue blocks, (c) performing mixed enzyme digestion and filtration through a screen, and collecting filtrate for centrifugation, (d) performing Percoll separation, (e) resuspending a cell sediment with a complete medium and inoculating into a culture flask, and (f) after culture for several days, replacing with a mixed complete medium for continuous culture. The method is simple to operate and efficiently obtains high activity and quick growing human renal tubular epithelial cells.

Description

technical field [0001] The invention belongs to the field of cell biology, in particular to a method for separating and culturing human renal tubular epithelial cells. Background technique [0002] Renal tubular epithelial cells are of scientific value for studying the pathogenesis of renal tubular-interstitial diseases, screening suitable therapeutic drugs, and fully understanding the biological characteristics and pathophysiological mechanisms of the cells. Therefore, providing a good human renal tubular epithelial cell model is very important. Since there are at least 15 to 20 types of cells in the mammalian kidney, the culture of renal tubular epithelial cells is relatively difficult compared to other tissue cells, mainly because it is difficult to separate and purify the same type of cells, and they are prone to degenerative development during subculture. At present, the research on renal tubular epithelial cells mostly adopts cell lines with low cultivation difficulty...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0686C12N2509/00
Inventor 不公告发明人
Owner JIANGYIN CHI SCI
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