Cell culture method for improving hepatocyte proliferation activity

A technology of proliferation activity and cell culture, which is applied in the field of cell culture to improve the proliferation activity of hepatocytes, can solve the problems of large differences in the real structure of the liver, and achieve the effect of good function and activity, and maintenance of function and activity.

Inactive Publication Date: 2016-04-13
FUZHOU HOSPITAL FOR INFECTIOUS DISEASE
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] The three-dimensional in vitro culture of hepatocytes is the main method of liver tissue engineering. Usually, polylactic-co-glycolic acid (PLGA), sodium alginate, chitosan, etc. are used as scaffold materials for three-dimensional culture of hepatocyte spheroids. Although this method It can maintain the long-term function and activity of liver cells in vitro, but it is quite different from the real structure of the liver in vivo

Method used

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  • Cell culture method for improving hepatocyte proliferation activity
  • Cell culture method for improving hepatocyte proliferation activity
  • Cell culture method for improving hepatocyte proliferation activity

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Experimental program
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Effect test

Embodiment 1

[0031] Hepatocytes (LO2 cell line, purchased from the Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) were cultured in complete medium to 80%-90% confluence, removed the complete medium, washed once with 0.01 MPBS, and washed once with 0.25% pancreatic Enzyme digestion for 2 minutes; add an equal volume to stop, after centrifuging at 300g for 5 minutes, collect the cells and add co-cultivation medium to adjust the cell solubility to 2.8×10 6 individual / mL.

Embodiment 2

[0033] Vascular endothelial cells (HUVECs, Basic Medical Cell Center, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences) were cultured in complete medium until they reached 80%-90% confluence, removed the complete medium, washed once with 0.01MPBS, and digested with 0.25% trypsin 2 minutes; add an equal volume of complete medium to stop, after centrifuging at 300g for 5 minutes, collect the cells and add co-cultivation medium to adjust the cell solubility to 0.6×10 6 individual / mL.

Embodiment 3

[0035] Human adipose stem cells (Kunming Cell Bank, Type Culture Collection Committee, Chinese Academy of Sciences) were cultured in complete medium to 80%-90% confluence, removed the complete medium, washed once with 0.01MPBS, and digested with 0.25% trypsin for 2 minutes; Add an equal volume of complete medium to stop, after centrifuging at 300g for 5 minutes, collect the cells and add co-cultivation medium to adjust the cell solubility to 0.6×10 6 individual / mL.

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Abstract

The invention relates to a cell culture method for improving the hepatocyte proliferation activity. The method comprises the following step: co-culturing hepatocyte, vascular endothelial cells and adipose-derived stem cells in a co-culture medium for 3 days or over 3 days according to a ratio of 14: (2 to 4): (2 to 4). The cell culture method disclosed by the invention has the beneficial effect mainly reflected in that a real structure and function of hepatocyte in vivo can be well simulated by adopting the cell culture method, and the activity of the cultured hepatocyte can well maintain the function and activity of the hepatocyte in vitro.

Description

(1) Technical field [0001] The invention relates to a cell culture method for improving the proliferative activity of hepatocytes. (2) Background technology [0002] Liver disease is a common disease in the human body, especially liver failure, which seriously endangers human health. Liver transplantation is an effective method to treat liver failure caused by hepatitis and cirrhosis, but the shortage of donor livers cannot meet the clinical needs. Liver tissue engineering is a discipline gradually formed under this background with the purpose of constructing a transplantable liver, which will bring hope for liver regeneration. The core of liver tissue engineering is to establish a three-dimensional complex of cells and biomaterials with vitality, to reconstruct the shape, structure and function of the damaged liver tissue, and to realize the permanent replacement of the liver. [0003] The three-dimensional in vitro culture of hepatocytes is the main method of liver tissue...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/0775
CPCC12N5/067C12N2502/1382C12N2502/28
Inventor 刘景丰刘小龙廖乃顺曾永毅蔡志雄王英超
Owner FUZHOU HOSPITAL FOR INFECTIOUS DISEASE
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