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Retina pigment epithelial cell culture medium and application thereof

A retinal pigment and epithelial cell technology, applied in the field of retinal pigment epithelial cell culture medium and culture medium, can solve the problems of limited number of RPE, slow RPE expansion speed, and inability to satisfy patients, so as to enhance vitality and prevent irregular aggregation , the effect of preventing oxidative damage

Inactive Publication Date: 2014-12-03
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although induced pluripotent stem cells (iPSC) do not involve ethical issues such as destroying embryonic cells, and avoid immune rejection, the possible tumorigenicity of iPSC limits its application in the treatment of AMD
[0004] At present, the number of fetal and adult RPE used to transplant RPE is extremely limited, which cannot meet the needs of patients
At the same time, under the previous RPE culture conditions, not only the use of expensive consumables such as transwell, but the RPE expansion speed is slow, and RPE monolayers with polarity and biological functions cannot be quickly formed.
These have seriously constrained the application of RPE cell replacement therapy

Method used

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  • Retina pigment epithelial cell culture medium and application thereof
  • Retina pigment epithelial cell culture medium and application thereof
  • Retina pigment epithelial cell culture medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The sources of reagents required in this embodiment are as follows:

[0044]

[0045] A culture medium for rapid in vitro expansion of embryonic retinal pigment epithelial cells is composed of a first RPE cell culture medium and a second RPE cell culture medium. Each component of the first RPE cell culture medium or the second RPE cell culture medium is filtered through a filter medium with a pore size of 0.22 μM before preparing the medium, and is used to remove possible contamination by filtering through a filter medium with a pore size of 0.22 μM bacteria. Its formula and preparation method are as follows:

[0046] 1) The first RPE cell culture medium (filtered through 0.22μM):

[0047]

[0048] 2) Second RPE cell culture medium (filtered through 0.22 μM):

[0049]

[0050] The above reagents should be mixed at room temperature and filtered through 0.22 μM. The above medium can be stored stably at 4°C for 3 months.

[0051] A kind of purposes of above-m...

Embodiment 2

[0060] A kind of retinal pigment epithelial cell culture medium, comprises the first RPE cell culture medium and the second RPE cell culture medium:

[0061] The first RPE cell culture medium comprises the following components, in 500ml, the consumption relationship of each component is as follows:

[0062] α Modified Minimal Essential Medium: 380ml, N1 Supplement: 5ml, Glutamine: 5mL, Penicillin-Streptomycin: 5mL, Non-Essential Amino Acids: 5mL, Taurine: 100mg, Hydrocortisone: 10μg, Triiodine Thyronine: 0.008μg, heat-inactivated fetal bovine serum: 100ml;

[0063] The second RPE cell culture medium comprises the following components, in 500ml, the consumption relationship of each component is as follows:

[0064] α Modified Minimal Essential Medium: 430ml, N1 Supplement: 5ml, Glutamine: 5mL, Penicillin-Streptomycin: 5mL, Non-Essential Amino Acids: 5mL, Taurine: 75mg, Hydrocortisone: 5μg, Triiodide Thyronine: 0.003μg, heat-inactivated fetal bovine serum: 50ml.

[0065] Each...

Embodiment 3

[0075] A retinal pigment epithelial cell culture medium, including the first RPE cell culture medium and the second RPE cell culture medium;

[0076] The first RPE cell culture medium comprises the following components, in 500ml, the consumption relationship of each component is as follows:

[0077] α Modified Minimal Essential Medium: 405ml, N1 Supplement: 5ml, Glutamine: 5mL, Penicillin-Streptomycin: 5mL, Non-Essential Amino Acids: 5mL, Taurine: 150mg, Hydrocortisone: 15μg, Triiodine Thyronine: 0.012μg, heat-inactivated fetal bovine serum: 75ml;

[0078] The second RPE cell culture medium comprises the following components, in 500ml, the consumption relationship of each component is as follows:

[0079] α Modified Minimal Essential Medium: 455ml, N1 Supplement: 5ml, Glutamine: 5mL, Penicillin-Streptomycin: 5mL, Non-Essential Amino Acids: 5mL, Taurine: 100mg, Hydrocortisone: 10μg, Triiodine Thyronine: 0.008μg, heat-inactivated fetal bovine serum: 25ml.

[0080] Each compon...

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Abstract

The invention relates to a retina pigment epithelial cell culture medium and an application thereof. The retina pigment epithelial cell culture medium comprises a first RPE (Retinal Pigment Epithelium) cell culture medium and a second RPE cell culture medium, wherein the first RPE cell culture medium and the second RPE cell culture medium are respectively used in 1.5 weeks, a RPE monolayer cell needed for cell replacement therapy can be quickly cultured; and the cultured RPE is polygonal, embedded and polarly-arranged monolayer cell and has the normal biological functions. Compared with the prior art, the RPE monolayer cell needed for cell replacement therapy can be quickly cultured in 1.5 weeks under the precondition that the conventional laboratory consumables are used; and the retina pigment epithelial cell culture medium is relatively low in cost and high in proliferation speed.

Description

technical field [0001] The invention relates to a culture medium and its application, in particular to a retinal pigment epithelial cell culture medium and its application, belonging to the field of biomedicine. Background technique [0002] Age-related macular degeneration (AMD) is a fundus disease that occurs frequently in the elderly. The clinical manifestations of AMD are progressive loss of central vision, which causes reading and other behavioral disturbances. Previous studies have found that the formation of AMD is related to the decrease in the ability of retinal pigment epithelial cells (RPE) to phagocytize and digest the outer segmental disc membrane of the retinal pigment epithelium (RPE). Efflux is deposited on Bruch's membrane, forming drusen. Apart from cell replacement therapy, there is currently no effective way to reverse this degenerative process and restore vision. Cell replacement therapy is to peel off the diseased RPE cells of patients through surgic...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 薛志刚范国平
Owner TONGJI UNIV
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