Human orbit preadipocyte cell line, construction method and application

A technology of fat precursor cells and cell lines, which is applied in the field of human orbital fat precursor cell line and construction, and can solve the problem of lack of fat cell model and so on.

Pending Publication Date: 2021-10-12
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Abstract
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Problems solved by technology

[0004] The current adipocyte models mainly include 3T3-L1 or subcutaneous adipocytes, and there is a lack of adipocyte models for thyroid-related ophthalmopathy

Method used

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  • Human orbit preadipocyte cell line, construction method and application
  • Human orbit preadipocyte cell line, construction method and application
  • Human orbit preadipocyte cell line, construction method and application

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Embodiment

[0029] Such as figure 1 Shown is a flowchart of the method for constructing a human orbital adipose precursor cell line of the present invention. The human orbital adipose precursor cell line of the present invention is constructed by the following method.

[0030] 1) Isolation of primary human orbital adipocytes

[0031] Orbital adipose tissue comes from healthy people and patients with thyroid exophthalmos. Rinse the obtained tissue with normal saline for 1-2 times to remove blood, cut the tissue fully with sterilized scissors, and put collagenase in a 37-degree water bath Digest for 30 minutes. After digestion, pass through a 100um cell sieve, centrifuge at 600rpm for 5min, spread in a culture dish, and culture to obtain human orbital adipose precursor cells. In the present invention, the orbital adipose tissue derived from healthy people and patients with thyroid exophthalmos can be used to construct the human orbital adipose precursor cell line of the present invention...

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Abstract

The invention discloses a human orbit preadipocyte cell line, a construction method and application. The cell line is obtained by immortalizing human orbit preadipocytes. The construction method of the cell line comprises the following steps of step 1, separating and culturing human orbit preadipocytes from human orbit adipose tissues; step 2, transfecting human orbit preadipocytes with the vector connected with hTERT gene, so that the cells overexpress hTERT; and step 3, screening out human orbit preadipocytes with negative CD90 expression. The cell line can be used as a cell research model for drug screening.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a human orbital fat precursor cell line, a construction method and application. Background technique [0002] Thyroid associated ophthalmopathy (TAO) is an organ-specific autoimmune disease mainly caused by hyperthyroidism (hyperthyroidism, also known as Graves’ Disease, GD) with exophthalmos as an important sign. The pathogenesis of TAO is closely related to the inflammation and hyperplasia of orbital adipose tissue. The pathological changes of TAO cause elevated posterior ocular pressure, oppressing the optic nerve and causing optic neuropathy; protruding eyeballs expose the cornea and are vulnerable to damage. These lesions not only cause visual dysfunction and even blindness, but also cause deformed appearance and affect the social and psychological health of patients. The quality of life of patients with TAO is severely affected, comparable to that of patients with diabetes or...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/85C12Q1/02
CPCC12N5/0653C12N15/85G01N33/5005C12N2510/04
Inventor 张芳金其煌张玲
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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