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Regeneration method of cartilage.

A technology of cartilage and chondrocytes, applied in the field of regenerative medicine, can solve problems such as transient, tumorigenic, and short-term effects of growth factors, and achieve the effect of enhancing ability, reducing side effects, and lasting effects

Pending Publication Date: 2020-03-17
SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Abstract
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  • Application Information

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Problems solved by technology

[0002] At present, the method of cartilage regeneration mainly achieves the purpose of regeneration by adding growth factors in vitro to promote its growth and expansion, but the growth factors are uncertain and have risks such as tumorigenesis; the added growth factors need to be varied and must reach a certain concentration to meet the needs of cell growth ; There are many uncertainties in exogenous growth factors, and there are risks of tumorigenesis; the effect of growth factors is short-lived and transient; growth factors may also act on other cells at the same time, resulting in uncontrollable results

Method used

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  • Regeneration method of cartilage.
  • Regeneration method of cartilage.

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Embodiment 1

[0029] This embodiment provides a cartilage regeneration method, which is a cartilage regeneration method based on gene transfection. The regeneration method includes the step of transfecting the target gene hsa-miR-193b-3p into chondrocytes.

[0030] In the step of transfecting the target gene hsa-miR-193b-3p into the chondrocytes, the target gene is transfected into the chondrocytes by the lentiviral vector, and at 35-39° C. and 4-6% CO 2 Incubate for 48-72 hours under the incubator conditions.

[0031] The regeneration method also includes: the step of culturing the transfected chondrocytes in vitro. In the step of culturing the transfected chondrocytes in vitro, the chondrocytes after successful transfection are routinely cultured in vitro to form a perichondrium sheet, that is, to obtain a cartilage regeneration material.

[0032] The acquisition of the chondrocytes comprises the following steps:

[0033] Take the residual ear cartilage discarded during the operation, c...

Embodiment 2

[0036] This embodiment provides a cartilage regeneration method, which includes the step of transfecting the target gene hsa-miR-193b-3p into chondrocytes.

[0037] In the step of transfecting the target gene hsa-miR-193b-3p into chondrocytes, the target gene is transfected into human chondrocytes by a lentiviral vector, and at 37° C. and 5% CO 2 Incubate for 60 hours in an incubator.

[0038] The regeneration method also includes: the step of culturing the transfected chondrocytes in vitro.

[0039] In the step of culturing the transfected chondrocytes in vitro, the chondrocytes after successful transfection are routinely cultured in vitro for 4 weeks to form a perichondrium sheet, that is, to obtain cartilage regeneration material.

[0040] The target gene contains GFP fluorescence, which is used to detect whether the target gene is successfully transfected into the chondrocytes.

[0041] The acquisition of the chondrocytes comprises the following steps:

[0042] Take the...

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Abstract

The invention discloses a regeneration method of cartilage. The regeneration method comprises the step: transfecting a target gene hsa-miR-193b-3p into cartilage cells. According to the disclosed method, the target gene hsa-miR-193b-3p is successfully transfected into human cartilage cells and intervention is carried out from the cell source required by regeneration, so that cell amplification andextracellular matrix secretion capacity is enhanced and the side reactions are reduced as much as possible. Therefore, the cartilage cell characteristics are improved from the source and the purposeof efficiently promoting cartilage regeneration is achieved.

Description

technical field [0001] The invention relates to the technical field of regenerative medicine, in particular to a cartilage regeneration method based on gene transfection. Background technique [0002] At present, the method of cartilage regeneration mainly achieves the purpose of regeneration by adding growth factors in vitro to promote its growth and expansion, but the growth factors are uncertain and have risks such as tumorigenesis; the added growth factors need to be varied and must reach a certain concentration to meet the needs of cell growth ; There are many uncertainties in exogenous growth factors, and there are risks of tumorigenesis; the effect of growth factors is short-lived and transient; growth factors may also act on other cells at the same time, resulting in uncontrollable results. [0003] Therefore, there is an urgent need to provide a cartilage regeneration method based on gene transfection to solve the above problems and achieve the purpose of efficientl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/867
CPCC12N5/0654C12N15/86C12N2510/00C12N2740/15043C12N2500/84
Inventor 陈霞张如鸿
Owner SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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