Human dental pulp stem cell culture medium and preparation method thereof

A dental pulp stem cell and culture medium technology, applied in the field of stem cell culture, can solve problems such as increasing the risk of dental pulp stem cells, and achieve the effects of increasing the expansion speed, increasing proliferation, and promoting proliferation

Inactive Publication Date: 2018-03-23
重庆斯德姆生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the culture system of dental pulp stem cells mainly adopts the medium containing animal-derived fetal bovine serum, whi

Method used

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  • Human dental pulp stem cell culture medium and preparation method thereof
  • Human dental pulp stem cell culture medium and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A culture medium for human dental pulp stem cells, including basal medium and supplements; the basal medium is α-MEM medium; the supplements include: 6% human AB serum by volume, 5 mg / L taurine, Lipoic acid 0.1mg / L, grape seed extract 2mg / L, carboxymethyl chitosan 0.6g / L, cucurbitacin B 0.3mg / L, platelet-derived growth factor 1μg / L, epidermal growth factor 1μg / L, yam Saponin 0.6mg / L and choline chloride 50μM.

[0024] Wherein, the grape seed extract is prepared by the following method:

[0025] (1) Pulverize the grape seeds, add 32% water and 0.03% yeast by weight of the grape seeds, seal and ferment at room temperature for 10 days, and then vacuum-dry until the water content is ≤7%;

[0026] (2) Using absolute ethanol as the extractant, perform subcritical extraction on the resultant of step (1), the extraction temperature is 60° C., the extraction pressure is 3.8 MPa, and the extraction time is 1 h to obtain the extract;

[0027] (3) The extract is concentrated unde...

Embodiment 2

[0030] A culture medium for human dental pulp stem cells, including basal medium and supplements; the basal medium is α-MEM medium; the supplements include: human AB serum with a volume percentage of 12%, taurine 10mg / L, Lipoic acid 0.2mg / L, grape seed extract 5mg / L, carboxymethyl chitosan 1.5g / L, cucurbitacin B 0.8mg / L, platelet-derived growth factor 4μg / L, epidermal growth factor 4μg / L, yam Saponin 1.8mg / L and choline chloride 65μM.

[0031] Wherein, the grape seed extract is prepared by the following method:

[0032] (1) Pulverize the grape seeds, add 35% water and 0.08% yeast by weight of the grape seeds, seal and ferment at room temperature for 12 days, and then vacuum-dry until the water content is ≤7%;

[0033] (2) Using absolute ethanol as the extractant, perform subcritical extraction on the resultant of step (1), the extraction temperature is 62°C, the extraction pressure is 4.8MPa, and the extraction time is 1.5h, to obtain the extract;

[0034] (3) The extract is...

Embodiment 3

[0037] A culture medium for human dental pulp stem cells, including basal medium and supplements; the basal medium is α-MEM medium; the supplements include: human AB serum with a volume percentage of 8%, taurine 6mg / L, sulfur Caprylic acid 0.12mg / L, grape seed extract 3mg / L, carboxymethyl chitosan 1.0g / L, cucurbitacin B 0.4mg / L, platelet-derived growth factor 2μg / L, epidermal growth factor 2μg / L, diosgenin 0.8mg / L, choline chloride 55μM.

[0038] Wherein, the grape seed extract is prepared by the following method:

[0039] (1) Pulverize the grape seeds, add 32% water and 0.03% yeast by weight of the grape seeds, seal and ferment at room temperature for 12 days, and then vacuum-dry until the water content is ≤7%;

[0040] (2) Using absolute ethanol as the extractant, perform subcritical extraction on the resultant of step (1), the extraction temperature is 60° C., the extraction pressure is 3.8 MPa, and the extraction time is 1.5 h to obtain the extract;

[0041](3) The extra...

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Abstract

The invention discloses a human dental pulp stem cell culture medium and a preparation method thereof. The culture medium is prepared from a basic culture medium and additives, wherein the basic culture medium is an alpha-MEM culture medium, and the additives are prepared from human AB serum with the volume percentage as 6 to 12%, 5 to 10mg/L of taurine, 0.1 to 0.2mg/L lipoic acid, 2 to 5mg/L of grape seed extract, 0.6 to 1.5g/L of carboxymethyl chitosan, 0.3 to 0.8mg/l of cucurbitacin B, 1 to 4mu g/L of platelet derived growth factor, 1 to 4mu g/L of epidermal growth factor, 0.6 to 1.8mg/L ofdiosgenin and 50 to 65mu M of choline chloride. The human dental pulp stem cell culture medium disclosed by the invention is prepared from specific ingredients, all the ingredients has an synergisticeffect, a dental pulp cell amplification speed can be remarkably improved, multidirectional differentiation capacity of the dental pulp stem cells is not affected, meanwhile human immune rejection isavoided, and reliable seed cells can be provided for clinical tooth repair and regeneration.

Description

technical field [0001] The invention belongs to the technical field of stem cell culture, and in particular relates to a human dental pulp stem cell culture medium and a preparation method thereof. Background technique [0002] Stem cells are cells with self-replication and multi-directional differentiation capabilities, which can continuously self-renew and differentiate into one or more cells that constitute human tissues or organs under specific conditions, which can restore the tissue's ability to regenerate and restore its functions. Researchers discovered a new type of adult stem cell in the pulp tissue of exfoliated deciduous teeth, and named it deciduous dental pulp stem cells. So far, many experiments have proved that deciduous dental pulp stem cells have a high degree of self-renewal, clone formation and osteogenic differentiation. ability. [0003] Dental pulp stem cells of deciduous teeth have the following advantages: 1) It is convenient to obtain materials, an...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0664C12N2500/33C12N2500/34C12N2500/38C12N2500/76C12N2501/11C12N2501/135C12N2501/999
Inventor 胡小东王健赵翔熊华强宋彩霞王仁杰邱阳黄启程李国焱唐玲叶劲涛曹玉昊
Owner 重庆斯德姆生物技术有限公司
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