Stem cell culture medium and preparation method thereof

A culture medium and stem cell technology, applied in the field of stem cell culture medium and its preparation, can solve the problem of slow cell proliferation, shorten time, facilitate product standardization, and eliminate pathogen contamination.

Inactive Publication Date: 2017-05-10
叶宗耀
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, a serum-free stem cell culture medium has been developed, but

Method used

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  • Stem cell culture medium and preparation method thereof
  • Stem cell culture medium and preparation method thereof
  • Stem cell culture medium and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. A stem cell culture medium

[0029] The stem cell culture medium includes a basal medium and additives, and the components of the additives are calculated at the final concentration, as shown in Table 1.

[0030] Table 1: Final concentration of additives added to the basic medium

[0031] additive Final concentration Human albumin 1mg / mL Transferrin 8μg / mL Glucagon 0.12μg / mL Activated carbon 2.5μg / mL Fibroblast growth factor 13ng / mL Epidermal growth factor 7ng / mL Linoleic acid 3μM Quercetin 5μg / mL Platycodin D 20μg / mL Laminine 2mg / mL Adenosine Deaminase 3μg / mL Myristic acid 1.2μM

[0032] The basic medium is DMEM medium.

[0033] Preparation:

[0034] S1 Add human albumin, transferrin, glucagon, fibroblast growth factor, epidermal growth factor, linoleic acid, quercetin, platycodin D, laminin and myristic acid to the basic medium , Stir for 25 minutes, add adenosine deaminase, continue stirring for 30 minutes, add activated carbon, stir evenly, and let ...

Embodiment 2

[0036] Example 2. A stem cell culture medium

[0037] The stem cell culture medium includes a basal medium and additives, and the components of the additives are calculated at the final concentration, as shown in Table 2.

[0038] Table 2: Final concentration of additives added to the basic medium

[0039] additive Final concentration Human albumin 3mg / mL Transferrin 13μg / mL Glucagon 0.15μg / mL Activated carbon 3μg / mL Fibroblast growth factor 18ng / mL Epidermal growth factor 11ng / mL Linoleic acid 6μM Quercetin 9μg / mL Platycodin D 25μg / mL Laminine 5mg / mL Adenosine Deaminase 7μg / mL Myristic acid 1.6μM

[0040] The basic medium is F12 medium.

[0041] Preparation:

[0042] S1 Add human albumin, transferrin, glucagon, fibroblast growth factor, epidermal growth factor, linoleic acid, quercetin, platycodin D, laminin and myristic acid to the basic medium , Stir for 35 minutes, add adenosine deaminase, continue stirring for 40 minutes, add activated carbon, stir evenly, and let s...

Embodiment 3

[0044] Example 3. A stem cell culture medium

[0045] The stem cell culture medium includes a basal medium and additives, and the components of the additives are based on the final concentration, as shown in Table 3.

[0046] Table 3: Final concentration of additives added to the basic medium

[0047] additive Final concentration Human albumin 2mg / mL Transferrin 10μg / mL Glucagon 0.14μg / mL Activated carbon 2.8μg / mL Fibroblast growth factor 15ng / mL Epidermal growth factor 9ng / mL Linoleic acid 5μM Quercetin 6μg / mL Platycodin D 23μg / mL Laminine 4mg / mL Adenosine Deaminase 5μg / mL Myristic acid 1.4μM

[0048] The basic medium is F12 medium.

[0049] Preparation:

[0050] S1 Add human albumin, transferrin, glucagon, fibroblast growth factor, epidermal growth factor, linoleic acid, quercetin, platycodin D, laminin and myristic acid to the basic medium , Stir for 30 minutes, add adenosine deaminase, continue stirring for 36 minutes, add activated carbon, stir evenly, and let stand...

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Abstract

The invention belongs to the technical field of cell culture mediums, and particularly relates to a stem cell culture medium and a preparation method thereof. The stem cell culture medium provided by the invention comprises a basic culture medium and an additive, wherein counted by final concentration, the additive is prepared from 1 to 3mg/mL human serum albumin, 8 to 13mug/mL transferrin, 0.12 to 0.15mug/mL glucagon, 2.5 to 3mug/mL activated carbon, 13 to 18ng/mL fibroblast growth factor, 7 to 11ng/mL epidermal growth factor, 3 to 6 muM linoleic acid, 5 to 9mug/mL quercetin, 20 to 25mug/mL platycodin, 2 to 5mg/mL laminine, 3 to 7mug/mL adenosine deaminase and 1.2 to 1.6muM myristic acid. By adopting the stem cell culture medium provided by the invention, stem cells can be amplified rapidly, and the culture time of the stem cells is shortened greatly.

Description

Technical field [0001] The invention belongs to the technical field of cell culture media, and specifically relates to a stem cell culture medium and a preparation method thereof. Background technique [0002] Stem cells cannot survive in a simple basal medium, and certain trace nutrients and growth factors must be provided in cell culture to enable the cells to grow and maintain their growth state. Therefore, the basic medium used is often supplemented with serum, such as horse serum or fetal bovine serum; because serum is an extremely complex mixture composed of many different sizes of biomolecules, it can provide the growth factors and hormones required by cells in vitro , Binding protein, and provide protection; and the serum is also rich in mitotic factors, which is also conducive to cell line and primary culture. However, the serum culture medium has certain disadvantages, and there is no way to avoid the heterologous body in the animal serum. If the cells cultivated from ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0602C12N2500/05C12N2500/24C12N2500/30C12N2500/36C12N2501/11C12N2501/113C12N2501/115C12N2501/119C12N2501/335C12N2501/73
Inventor 叶宗耀
Owner 叶宗耀
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