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Serum-free medium for rapid generation of induced pluripotent stem cells

A serum-free medium and pluripotent stem cell technology, applied in the field of cell culture medium, can solve the problems of reduced application value and safety of induced pluripotent stem cells, inability to avoid animal serum heterogeneity, and slow cell proliferation, etc., to achieve Shorten the time, avoid the rejection phenomenon, and promote the effect of proliferation

Active Publication Date: 2017-10-20
GUANGZHOU RAINHOME PHARM&TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, culture medium containing fetal bovine serum is mostly used for the culture of induced pluripotent stem cells. The medium containing serum cannot avoid the heterogeneity in animal serum. Body rejection and conflict, which makes induced pluripotent stem cells in the clinical application value and safety reduced
Although a serum-free medium for induced pluripotent stem cells has been developed, there are problems such as slow cell proliferation

Method used

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  • Serum-free medium for rapid generation of induced pluripotent stem cells

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1, a kind of serum-free culture medium that induced pluripotent stem cells produce rapidly

[0032] The serum-free medium for rapid production of induced pluripotent stem cells includes basal medium and additives, and the components of the additives include: cell growth factor 20ng / mL, transferrin 21 μg / mL, fibronectin 6 μg in terms of final concentration / mL, polyvinylpyrrolidone 10 μg / mL, genistein 1 μg / mL, mannitol 4 μg / mL, insulin 5 μg / mL, L-glutamine 11 μg / mL, palmitic acid 12 μg / mL and hesperidin 8 μg / mL mL.

[0033] The basal medium is F12 medium.

[0034] The cell growth factor is composed of fibroblast growth factor, epidermal growth factor and transforming growth factor in a weight ratio of 8:5:3.

[0035] Preparation:

[0036] S1 Add cell growth factors, insulin, L-glutamine and palmitic acid to the basal medium, stir at a stirring speed of 80r / min for 35min, add transferrin, fibronectin, genistein and orange Dermoside, continue to stir for 40m...

Embodiment 2

[0038] Embodiment 2, a kind of serum-free culture medium that induced pluripotent stem cells produce rapidly

[0039] The serum-free medium for the rapid production of induced pluripotent stem cells includes basal medium and additives, and the components of the additives include: cell growth factor 40ng / mL, transferrin 30μg / mL, fibronectin 9μg in terms of final concentration / mL, polyvinylpyrrolidone 15 μg / mL, genistein 3 μg / mL, mannitol 7 μg / mL, insulin 8 μg / mL, L-glutamine 15 μg / mL, palmitic acid 16 μg / mL and hesperidin 15 μg / mL mL.

[0040] The basal medium is DMEM medium.

[0041] The cell growth factor is composed of fibroblast growth factor, epidermal growth factor and transforming growth factor in a weight ratio of 10:3:1.

[0042] Preparation:

[0043] S1 Add cell growth factors, insulin, L-glutamine and palmitic acid to the basal medium, stir at a stirring speed of 120r / min for 50min, add transferrin, fibronectin, genistein and orange Dermoside, continue to stir f...

Embodiment 3

[0045] Embodiment 3, a kind of serum-free culture medium that induced pluripotent stem cells produce rapidly

[0046] The serum-free medium for rapid production of induced pluripotent stem cells includes basal medium and additives, and the components of the additives include: cell growth factor 26 ng / mL, transferrin 24 μg / mL, fibronectin 7 μg in terms of final concentration / mL, polyvinylpyrrolidone 13 μg / mL, genistein 2 μg / mL, mannitol 5 μg / mL, insulin 7 μg / mL, L-glutamine 12 μg / mL, palmitic acid 14 μg / mL and hesperidin 11 μg / mL mL.

[0047] The basal medium is F12 medium.

[0048] The cell growth factor is composed of fibroblast growth factor, epidermal growth factor and transforming growth factor in a weight ratio of 9:4:2.

[0049] Preparation:

[0050] S1 Add cell growth factors, insulin, L-glutamine and palmitic acid to the basal medium, stir, the stirring speed is 90r / min, the stirring time is 44min, add transferrin, fibronectin, genistein and orange Dermoside, cont...

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Abstract

Belonging to the technical field of cell mediums, the invention in particular relates to a serum-free medium for rapid generation of induced pluripotent stem cells. The serum-free medium for rapid generation of induced pluripotent stem cells includes a basic medium and an additive, and the additive comprises the following components in terms of final concentration: 20-40ng / mL cell growth factor, 21-30microg / mL transferrin, 6-9microg / mL fibronectin, 10-15microg / mL polyvinylpyrrolidone, 1-3microg / mL genistein, 4-7microg / mL mannitol, 5-8microg / mL insulin, 11-15microg / mL L-glutamine, 12-16microg / mL palmitic acid, and 8-15microg / mL hesperidin. The serum-free medium for rapid generation of induced pluripotent stem cells provided by the invention has the advantages of clear components, no serum, high safety, and rapid generation of induced pluripotent stem cells.

Description

Technical field [0001] The invention belongs to the technical field of cell culture media, and specifically relates to a serum-free culture medium for rapid production of induced pluripotent stem cells. Background technique [0002] Induced pluripotent stem cells (IPS) were originally developed by Japanese scientist Shinya Yamanaka in 2006 by using a viral vector to transfer a combination of four transcription factors into differentiated somatic cells to reprogram them. They are similar to embryonic stem cells and embryonic APSC pluripotent cells. of a cell type. By introducing foreign genes, somatic cells are dedifferentiated into pluripotent stem cells, which we call induced pluripotent stem cells. Since the emergence of induced pluripotent stem cell technology, researchers around the world have been researching and developing this technology. It not only has the pluripotency of embryonic stem cells, but also does not involve human ethical issues. It is of immeasurable va...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10
CPCC12N5/0696C12N2500/25C12N2500/32C12N2500/35C12N2500/36C12N2500/50C12N2500/90C12N2501/11C12N2501/113C12N2501/115C12N2501/117C12N2501/119C12N2501/148C12N2501/15C12N2501/998C12N2501/999C12N2510/00
Inventor 车七石
Owner GUANGZHOU RAINHOME PHARM&TECH CO LTD
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