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Stem cell culture medium and application thereof and stem cell cultivation method

A stem cell and culture medium technology, applied in the field of cell culture, can solve the problems of increase and decrease of cell potential passage times, slow growth of stem cells, etc.

Active Publication Date: 2013-04-24
苏州博棠再生医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a stem cell culture medium and its application and stem cell culture method, aiming to solve the problem of slow growth and cell potentiality in the culture of stem cells in the existing culture medium supplemented with bovine serum. The problem of greatly reducing the number of times

Method used

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  • Stem cell culture medium and application thereof and stem cell cultivation method
  • Stem cell culture medium and application thereof and stem cell cultivation method
  • Stem cell culture medium and application thereof and stem cell cultivation method

Examples

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Effect test

Embodiment 1

[0182] Example 1. Acceleration of stem cell proliferation

[0183] The conventional medium formula for culturing stem cells is: DMEM / F12 basic medium, 10% fetal bovine serum (FBS). In order to compare the culture medium described in the present invention and to examine the influence of the culture medium described in the present invention on the expansion speed of human stem cell culture, human stem cells from three kinds of tissue and organ sources: fat, bone marrow and umbilical cord were selected in this example. (ADSC, BMSC, UMBSC) for in vitro cell culture experiments. The cell seeding density is controlled between 10-20%. Before seeding, the culture dish is treated with cell adhesion material at room temperature for 2 hours or incubated overnight at 4°C. After seeding, culture medium is added (37°C, 5% CO 2 ). Cell density was measured every 12 hours by the MTT assay. The test results are shown in Figure 1~ image 3 As shown, the stem cells (solid column) cultured in...

Embodiment 2

[0184] Example 2. Potential analysis of proliferating mesenchymal stem cells into chondrogenic cells in vitro

[0185] In order to examine the influence of stem cells on their differentiation potential after being amplified by the medium described in the present invention for 10 generations, human stem cells derived from fat, bone marrow and umbilical cord were measured in this example according to the method described in the literature ( ADSC, BMSC, UMBSC) after expansion of chondrogenic differentiation assay (Estes BT et al. Nat Protocol 2010; 5 : 1294–1311; Awad HA et al., Biomaterials. 2004 Jul;25(16):3211-22; Cheng NC et al., Tissue Eng Part A. 2009 Feb;15(2):231-41). When the stem cells grow to a suitable density (cell density 20-80%), add chondrogenic induction medium (DMEM / F12 medium with 1% FBS, 10 mg / L TGF-β1, 50 nM vitamin C, 6.25 mg / L insulin , 1% double antibody), the medium was changed once a day, and the total induction was 2-3 weeks. Alcian blue staining w...

Embodiment 3

[0186] Example 3. Potential analysis of adipogenic cells

[0187]In order to further examine the influence of stem cells on their differentiation potential after being amplified by the medium described in the present invention for 10 generations, in this example, we followed the adipogenic induction and oil red O staining methods of adipose-derived stem cells (ADSC) described in the literature. The differentiation experiment of adipocytes after the expansion of human stem cells (ADSC, BMSC, UMBSC) derived from fat, bone marrow and umbilical cord three kinds of tissues and organs was measured according to the method described in the literature (Yu G et al., Methods in Molecular Biology, 2011, Volume 702, Part 3, 193-200; Bunnell BA et al., Methods. Volume 45, Issue 2, June 2008, Pages 115–120; Gimble JM & Guilak F. Cytotherapy. 2003, Vol.5, No. 5, Pages 362-369). The specific method is, when the stem cell density is about 80%, replace it with adipogenic medium for induction. ...

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Abstract

The invention discloses a stem cell culture medium, an application of the stem cell culture medium and a stem cell cultivation method. Blood serum does not exist in the stem cell culture medium. The stem cell culture medium comprises amino acid, vitamin, salt, lipoid, cytokines and egg white polypeptide. The stem cell culture medium is suitable for fast cultivating stem cells which are tissue sources of human and mammal, and comprises but not is limited by fat mesenchyme stem cells, mesenchymal stem cells and umbilical cord blood stem cells. The culture medium enables increasing speed of the cells to be improved by 3-5 times, and differential potentials of the cells can not be affected. Compared with an ordinary stem cell culture medium, stem cells from different sources can be fast expanded, passage number is prolonged, and proficiency properties of the stem cells can be well kept.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a stem cell culture medium, its application and a stem cell culture method. Background technique [0002] Many studies have shown that stem cells can not only self-renew, but also differentiate into other functional cells under suitable conditions. Therefore, stem cells are expected to become an effective means of treating difficult human diseases. However, the content of stem cells in normal adult tissues is very small. How to rapidly expand and cultivate stem cells in vitro is an important technology for studying the mechanism of stem cells and exploring their therapeutic methods in the treatment of human diseases. [0003] The content of stem cells is as small as possible, but they are widely distributed in various tissues and organs of mammals, including but not limited to bone marrow, umbilical cord, adipose tissue, brain tissue, retina, heart, liver, lung and skin. A large numb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/0775C12N5/0789
CPCC12N5/0037C12N5/0663C12N5/0665C12N5/0667C12N2500/32C12N2500/33C12N2500/36C12N2500/38C12N2501/105C12N2501/11C12N2501/113C12N2501/115C12N2501/125C12N2501/135C12N2501/165C12N2501/235C12N2501/39C12N2501/91
Inventor 刘小青应明陈光风郑龙坡
Owner 苏州博棠再生医学科技有限公司
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