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Cell Anti-Stress Gene Fragments and Modified Hepg2 Cells

A technology of gene fragments and cells, which is applied in the field of cell anti-stress gene fragments and transformed HepG2 cells, can solve the problems of cell nucleus proliferation, cell dehydration and apoptosis, and achieve the effect of improving cell stress resistance

Active Publication Date: 2022-07-05
WUHAN TOGO MEDITECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] When the existing HepG2 cells are used for blood purification, in the environment of high-concentration toxin plasma, it is easy to proliferate the nucleus, and the cells will have the defect of dehydration and apoptosis

Method used

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  • Cell Anti-Stress Gene Fragments and Modified Hepg2 Cells
  • Cell Anti-Stress Gene Fragments and Modified Hepg2 Cells
  • Cell Anti-Stress Gene Fragments and Modified Hepg2 Cells

Examples

Experimental program
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Effect test

Embodiment

[0028] Example: Protocol for HepG2 cell line transformation:

[0029] 1) The whole genome was assembled into pTBSCH3-CPS1-OATP1B3-G418 and linearized, the sequence is shown in SEQ No. 2.

[0030] 2) Digest HepG2 (ATCC, American Type Bacteria Collection Center) that night, and its sequence can be found in (NCBIGenBank). Cells were seeded into T25 at a density of 40%-50%, and after overnight adherence, the next day Transfection was performed in the morning.

[0031] 3) Transfection reagents and materials

[0032]

[0033] (plasmid:transposon = 10:1)

[0034] Mix well, let stand for 5 min, add to T25, and change the medium according to the cell state.

[0035] 4) Five days after the cells were transfected, they were digested and counted, and about 1000-2000 cells per well were seeded into a 96-well plate.

[0036] 5) After the cells grow enough to a certain number, continue to seed plate screening. After groping for negative cells (HepG2) in the early stage, screening was...

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Abstract

The invention discloses a cell anti-stress gene fragment and a modified HepG2 cell, wherein the cell anti-stress gene fragment is composed of two gene fragments, CPS1 and OATP1B3 in sequence. The HepG2 cells transformed with this gene fragment increased the secretion of the cell growth factor ALR by 5%, the secretion of the hepatocyte growth factor HGF doubled, and the recovery speed was increased by 10 times after the reaction to promote liver injury, and the cell resistance to stress was greatly improved.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a cell anti-stress gene fragment and a modified HepG2 cell. Background technique [0002] HepG2 cells are a type of liver cancer cells. The cells secrete a variety of plasma proteins: albumin, α2-macroglobulin, plasminogen, transferrin, etc. [0003] When the existing HepG2 cells are used for blood purification, they are prone to proliferation of nuclei in the environment of high concentration of toxin plasma, and the cells will have defects of dehydration and apoptosis. SUMMARY OF THE INVENTION [0004] The purpose of the present invention is to overcome the defects of the existing HepG2 cells, and to provide a cell anti-stress gene fragment and a modified HepG2 cell. [0005] In order to achieve the above purpose, the present invention first provides a cell anti-stress gene fragment, which is composed of two gene fragments, CPS1 and OATP1B3, in sequence, wherein the secre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/62C12N15/85C12N15/65C12N5/10C12R1/91
CPCC12N9/93C12N15/85C12N15/65C07K14/47C12Y603/05005C07K2319/00
Inventor 望威王柯丰明乾
Owner WUHAN TOGO MEDITECH CO LTD
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