Dental pulp stem cell culture medium

A dental pulp stem cell and culture medium technology, applied in the field of stem cell culture, can solve the problems of dental pulp stem cell culture difficulties, containing heterologous animal protein, and reduced differentiation potential, and achieve high scientific research and medical application value

Inactive Publication Date: 2015-06-17
ANHUI NEW LIFE STEM CELL TECH CO LTD
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AI Technical Summary

Problems solved by technology

[0005] In order to overcome the above-mentioned deficiencies in the prior art, the present invention provides a dental pulp stem cell culture medium, which can solve the difficulties in the cultivation of dental pulp stem cells caused by backward dental pulp stem cell culture methods, reduced differentiation potential, and heterogeneous animal culture medium. protein problem

Method used

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  • Dental pulp stem cell culture medium

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Effect test

Embodiment 1

[0010] Experiment on accelerating the proliferation of dental pulp stem cells

[0011] 1. Test medium: conventional medium components: DMEM minimal medium, 10% fetal bovine serum (FBS); the medium of the present invention includes: SITE100* (sigma) 10ml, ascorbic acid 300mM, PDGF 5ug, hydrogenated Pine 5ug, EGF 3ng, b-FGF, 3ng, PTH 100ng, dexamethasone 10mM, IMDM balance.

[0012] 2. Source of cultured stem cells: dental pulp stem cells derived from dental pulp

[0013] 3. In vitro cell culture experiment: the cell seeding density is 4000 cells / ml, and the cells are seeded in a 12-well plate and placed in an incubator (37 degrees, 5% CO2). The number of cells was measured with a hemocytometer every 24 hours, and it was found that the dental pulp stem cells cultured in the medium of the present invention showed a faster expansion rate than the cells cultured in the conventional medium (data are three independent The average value of the experiment).

[0014]

Embodiment 2

[0016] The effect of analysis on the potential of proliferated dental pulp stem cells to differentiate into osteoblasts, chondroblasts and adipocytes in vitro

[0017] 1. Test medium: the medium of the present invention.

[0018] 2. Source of cultured cells: dental pulp stem cells derived from dental pulp

[0019] 3. In vitro cell differentiation experiment: the cells were passaged and expanded to the P10 generation. According to the instructions, the P10 generation dental pulp stem cells were tested for differentiation using LONZA's adipogenic, osteogenic, and cartilage detection kits. The test results showed that the description of the present invention The ratio of dental pulp stem cells cultured in the culture medium to differentiate into adipocytes, chondroblasts, and osteoblasts is above 80% (data is the average of three independent experiments). Therefore, the culture medium described in the present invention can effectively maintain the multidirectional differentiation abili...

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Abstract

The invention discloses a dental pulp stem cell culture medium. IMDM is taken as a basal culture medium, SITE100* and ascorbic acid 200-400 mM are included in 1 L of culture medium, the SITE serves as an indispensable component for cell growth, the ascorbic acid and fibronectin are helpful to a dental pulp stem cell to form cell epimatrix, and are favorable for the growth of the dental pulp stem cell. The dental pulp stem cell culture medium comprises 1 to 10 ug of PDGF, 1 to 10 ug of hydrocortisone, 1 to 5 ng of EGF, 1 to 5 ng of b-FGF1, 50 to 200 ng of PTH, and 1 to 20 mM of dexamethasone, the combination of all growth factors can effectively enhance the multiplication capacity of the dental pulp stem cell, and can keep the undifferentiated state of the cell. The culture medium provided by the invention can quickly expand and increase the dental pulp stem cell and keep the multi-directional differentiation capability of the dental pulp stem cell, reaches an expanding and increasing speed which is increased by 2 to 3 times by compared with that of a conventional culture medium, and has very high scientific research and clinical application values.

Description

Technical field [0001] The invention relates to the technical field of stem cell culture, in particular to a dental pulp stem cell culture medium. Background technique [0002] Stem Cells (Stem Cells, SC) are a type of self-renewing multipotent cells, which are primitive and unspecialized cells. Under certain conditions, it can differentiate into a variety of functional cells, with the potential to regenerate various tissues and organs and the human body. Stem cells exist in all multicellular tissues, can divide into a variety of specialized cells through mitosis and differentiation, and can use self-renewal to provide more stem cells. There are many sources of stem cells, including bone marrow, umbilical cord, cord blood, teeth and fat. [0003] Dental pulp stem cells were first discovered in 2000. Gronthos et al. found that dental pulp stem cells exist in both isolated deciduous teeth and wisdom teeth. This is a type of cell with high proliferation ability, high self-renewal a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074
Inventor 杨茜董成友张路军
Owner ANHUI NEW LIFE STEM CELL TECH CO LTD
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