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A kind of in vitro culture method of minipig islet cells

A technology of islet cell and in vitro culture, applied in cell culture active agents, pancreatic cells, artificial cell constructs, etc., can solve the problems of high apoptosis rate, poor stability, low cell viability, etc. rate, significant market value and social value

Active Publication Date: 2022-03-29
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing in vitro culture technology of minipig islet cells has the defects of high apoptosis rate, low cell viability and poor stability

Method used

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  • A kind of in vitro culture method of minipig islet cells

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Embodiment 1

[0033] A method for culturing minipig islet cells in vitro, the steps are as follows:

[0034] 1) Prepare a cell growth solution, the cell growth solution includes the first base solution composed of calf serum, F12 culture medium and DMEM culture medium in a volume ratio of 5:100:25, and add Apoptosis inhibitor Z-VAD-FMK 8μM (based on the total molar mass of all substances in the cell growth medium), insulin-like growth factor 12μg / L, vascular endothelial growth factor 5μg / L, fibroblast growth factor 2μg / L, TGF- β 6 μg / L, insulin 12 μg / L, penicillin 90 U / mL, streptomycin 90 U / mL, transferrin 3 μg / L, first additive 25 μg / L;

[0035] 2) Prepare a cell culture solution, the cell culture solution includes a second base solution consisting of calf serum, F12 culture medium and DMEM culture medium in a volume ratio of 3:90:30, and add Apoptosis inhibitor Z-VAD-FMK13μM, insulin-like growth factor 6μg / L, vascular endothelial growth factor 2μg / L, fibroblast growth factor 2μg / L, TGF-β...

Embodiment 2

[0050] A method for culturing minipig islet cells in vitro, the steps are as follows:

[0051] 1) Prepare a cell growth solution, the cell growth solution includes the first base solution composed of calf serum, F12 culture medium and DMEM culture medium in a volume ratio of 5:100:25, and add Apoptosis inhibitor Z-VAD-FMK9 μM, insulin-like growth factor 13 μg / L, vascular endothelial growth factor 6 μg / L, fibroblast growth factor 4 μg / L, TGF-β 8 μg / L, insulin 13 μg / L, penicillin 90 U / mL, streptomycin 90U / mL, transferrin 3.5μg / L, first additive 28μg / L;

[0052] 2) Prepare a cell culture solution, the cell culture solution includes a second base solution consisting of calf serum, F12 culture medium and DMEM culture medium in a volume ratio of 3:90:30, and add Apoptosis inhibitor Z-VAD-FMK 14 μM, insulin-like growth factor 8 μg / L, vascular endothelial growth factor 3 μg / L, fibroblast growth factor 4 μg / L, TGF-β 13 μg / L, insulin 9 μg / L, penicillin 90 U / mL, streptomycin 90U / mL, ...

Embodiment 3

[0067] A method for culturing minipig islet cells in vitro, the steps are as follows:

[0068] 1) Prepare a cell growth solution, the cell growth solution includes the first base solution composed of calf serum, F12 culture medium and DMEM culture medium in a volume ratio of 5:100:25, and add Apoptosis inhibitor Z-VAD-FMK 10 μM, insulin-like growth factor 15 μg / L, vascular endothelial growth factor 8 μg / L, fibroblast growth factor 5 μg / L, TGF-β 10 μg / L, insulin 15 μg / L, penicillin 90 U / mL, streptomycin 90U / mL, transferrin 4μg / L, first additive 30μg / L;

[0069] 2) Prepare a cell culture solution, the cell culture solution includes a second base solution consisting of calf serum, F12 culture medium and DMEM culture medium in a volume ratio of 3:90:30, and add Apoptosis inhibitor Z-VAD-FMK16 μM, insulin-like growth factor 10 μg / L, vascular endothelial growth factor 4 μg / L, fibroblast growth factor 5 μg / L, TGF-β 15 μg / L, insulin 10 μg / L, penicillin 90 U / mL, streptomycin 90U / mL...

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Abstract

The invention discloses a method for culturing miniature pig islet cells in vitro. The method comprises the following steps: configuring cell growth liquid; tube, centrifuge, discard the supernatant, use the cell growth medium to suspend the cells at the bottom of the centrifuge tube, then inhale into the first petri dish containing the cell growth medium, and put it into the incubator for cultivation; after culturing, Replace the culture dish and medium, and use the second culture dish containing cell culture medium for cultivation, and then replace the culture dish and medium every 1-3 days, and use cell culture medium as the medium. The invention can reduce the apoptosis rate of cells, improve the vitality of pancreatic islet β cells, and the cultured cells have sensitive glucose stimulation response, can meet the requirements of researching human pancreatic islet diseases, and have important market value and social value.

Description

technical field [0001] The invention relates to the technical field of animal cell culture, in particular to a method for culturing minipig islet cells in vitro. Background technique [0002] As the earliest animals raised by humans, pigs have the same omnivorous behavior as humans, but are quite different from dogs (carnivorous) and monkeys (vegetarian). Its physiological function, material metabolism, organ morphology and disease mechanism are very similar to human beings. Pigs and humans have a similar cytochrome oxidase P450 system. The important metabolic enzymatic characteristics determine that the biotransformation of drugs in the body is very similar to that of humans. It is an ideal model animal for drug safety evaluation and screening, and human diseases, especially in anti- Oncology drugs, cardiovascular system drugs, skin-penetrating drugs, non-steroidal antibiotics, and digestive system drugs have shown obvious advantages over dogs and non-human primates. Coup...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0676C12N2500/25C12N2500/76C12N2501/115C12N2501/105C12N2501/734C12N2501/165C12N2501/15
Inventor 谢水林黄黎珍邹芬
Owner SOUTH CHINA UNIV OF TECH
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