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Serum-free culture medium for dental pulp stem cells

A technology of serum-free medium and dental pulp stem cells, which is applied in the field of stem cell culture, can solve problems such as unclear components, and achieve the effects of clear components, cost saving, and easy operation

Inactive Publication Date: 2019-07-12
浙江优牙生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

FBS has unclear components and contains heterogeneous proteins, which may carry the risk of bacteria, viruses, protein infectious diseases or prions and may cause immune reactions, while the serum-free medium of this patent has clear components, no animal-derived components, and can Maintain cell viability for a long time

Method used

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  • Serum-free culture medium for dental pulp stem cells
  • Serum-free culture medium for dental pulp stem cells
  • Serum-free culture medium for dental pulp stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 A kind of serum-free culture medium

[0027] (Homemade) A serum-free medium for dental pulp stem cells includes growth factors, hormones, proteins, vitamins and reducing substances, substances that promote adhesion, enzyme inhibitors and various trace elements, a-MEM and double antibodies (100 U / mL penicillin+100 ug / mL streptomycin);

[0028]Growth factors include PDGF-AB: 21.45-60.49ng / mL, PDGF-BB: 13.57-32.63ng / mL, FGF: 3.76-14.5ng / mL, EGF: 3-6.12ng / mL, TGF-β1: 7.45-13.84 ng / mL, bFGF: 40.71-97.89 ng / mL;

[0029] Hormones include insulin: 0.1-10μg / ml;

[0030] Proteins include globulin: 1.8-2g / dL, albumin: 3.0-3.6g / dL;

[0031] Vitamin amino acids include choline chloride 0.5-0.8 mg / dL, D-calcium pantothenate 1-3 mg / L, folic acid 0.1-3 mg / L, nicotinamide 0.2-0.3 mg / dL, pyridoxine hydrochloride 0.01-0.03 mg / dL, riboflavin 0.1-0.5 mg / L, vitamin B35-10 mg / dL, vitamin B6 0.1-1mg / dL, vitamin B12 0.1-0.8 mg / L, inositol 1-3 mg / dL, glycine 2-4mg / ...

Embodiment 2

[0036] Embodiment 2 A kind of serum-free culture medium

[0037] (TBD-2) A serum-free medium for dental pulp stem cells including 10% serum-free supplement (purchased from Tianjin Haoyang Huake Biotechnology Co., Ltd.), 90% basal medium (purchased from Tianjin Haoyang Huake Biotechnology Co., Ltd. company) and double antibody (100 U / mL penicillin + 100 ug / mL streptomycin)

[0038] Serum-free supplements include recombinant human serum albumin, recombinant human transferrin, recombinant human insulin, rhEGF, rhbFGF, rhPDGF-BB, rhSCF, and fibronectin

[0039] Basal medium includes amino acids, vitamins, inorganic salts, sodium selenite, glucose, phenol red, and sodium pyruvate

[0040] Amino acids include glycine, L-alanine, L-arginine hydrochloride, L-asparagine, L-aspartic acid, L-cystine hydrochloride, L-glutamic acid, L-group Amino acid hydrochloride, L-isoleucine, L-leucine, L-lysine hydrochloride, L-methionine, L-phenylalanine, L-proline, L-serine, L - Threonine, L-Tryp...

Embodiment 3

[0043] Embodiment 3 A kind of serum-free culture medium

[0044] (YC-1) A serum-free medium for dental pulp stem cells including serum-free medium (purchased from Youkang Hengye Biotechnology Co., Ltd.) and double antibody (100 U / mL penicillin + 100 ug / mL streptomycin)

[0045] Serum-free medium includes amino acids: glycine, L-alanine, L-arginine hydrochloride, L-asparagine, L-aspartic acid, L-cysteine ​​hydrochloride, L-glutamine amino acid, L-glutamine, L-histidine hydrochloride, L-isoleucine, L-leucine, L-lysine hydrochloride, L-methionine, L-phenylalanine , L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine and L-valine. Vitamins include choline chloride, calcium D-pantothenate, folic acid, niacinamide, pyridoxine hydrochloride, riboflavin, thiamine hydrochloride, vitamin B12, and inositol. Inorganic salts include: calcium chloride, magnesium chloride, magnesium sulfate (7H2O), potassium chloride, sodium chloride, disodium hydrogen phosphate (7H2O), sodium dihyd...

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Abstract

The invention provides a serum-free culture medium for dental pulp stem cells. The serum-free culture medium for the dental pulp stem cells is characterized in that no animal source exists, the composition is clear, and various nutrient additives can replace FBS, so that the cost is reduced, the source is natural, the culture medium is rich in various growth factors, hormones proteins and the likewhich are required for stem cell growth, the most natural cell growth condition can be provided, adherent treatment is not needed, and the operation is simple.

Description

technical field [0001] The invention relates to the technical field of stem cell culture, in particular to a serum-free medium for dental pulp stem cells. Background technique [0002] In 2000, Gronthos et al [Gronthos SM, Brabim J.Postnatal human dental pulpstem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci U S A, 2000, 97(25): 13625-13630. Adult stem cells with clonal properties are isolated from the pulp, named dental pulp stem cells. Dental pulp stem cells (Dental pulp stem cells, DPSCs) are easy to obtain, rich in sources, without ethical and moral restrictions, low immunogenicity and good multi-directional differentiation potential, and have great application potential in the fields of stem cell therapy and regenerative medicine [N. Nuti, C. Corallo, B.M. Chan, et al. Multipotent differentiation of human dental pulp stem cells: a literature review. StemCell Rev. 12 (2016) 511-523.]. However, a certain proportion of animal-derived serum (such as fetal bovine ...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0664C12N2500/25C12N2500/90C12N2501/11C12N2501/115C12N2501/135C12N2501/15C12N2501/998
Inventor 叶青松郑平平潘婷贺捷
Owner 浙江优牙生物科技有限公司
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