Clinical-grade human mesenchymal stem cell serum-free complete medium

A complete culture medium and mesenchymal stem cell technology, applied in animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve the problems of cell pollution, increase of pathogens, high price, etc., to promote cell growth, promote cell proliferation, Effect of reducing toxicity

Active Publication Date: 2013-08-14
QINGDAO RESTORE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is a risk of potential virus infection in animal-derived serum, and the serum components are complex, it is not easy to qualitatively study the effect of each component on the growth and differentiation of stem cells, and it is easy to cause allergic reactions
[0004] At present, there are many mesenchymal stem cell culture medium products on the market. Disadvantages: 1. The use of animal-derived serum increases pathogens, heterologous antigens, and instability between batches
The US FDA is considering banning the use of fetal bovine serum for cell therapy; 2. Stem cells are easy to differentiate and cannot maintain stemness f

Method used

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  • Clinical-grade human mesenchymal stem cell serum-free complete medium
  • Clinical-grade human mesenchymal stem cell serum-free complete medium
  • Clinical-grade human mesenchymal stem cell serum-free complete medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 A clinical-grade human mesenchymal stem cell serum-free complete medium provided by the present invention. The composition and concentration are shown in Table 1. The following components are dissolved and mixed according to their respective characteristics, and the volume of HEPES is fixed, 0.22μm Sterilize through filtration membrane and store at 4°C.

[0035] Table 1 Composition and concentration of the clinical grade human mesenchymal stem cell serum-free complete medium of the present invention

[0036]

[0037]

Embodiment 2

[0038] Example 2 The clinical grade human mesenchymal stem cell serum-free complete medium of the present invention has been tested by a third-party, and all indicators have reached the standard, as shown in Table 2:

[0039] Table 2 The third-party quality test items and results of the clinical grade human mesenchymal stem cell serum-free complete medium of the present invention

[0040]

[0041] It can be seen from Table 2 that the clinical grade human mesenchymal stem cell serum-free complete medium of the present invention has all the test indicators qualified. Only qualified media can be used in clinical practice.

Embodiment 3

[0042] Example 3. Experiment of culturing human mesenchymal stem cells using the clinical grade human mesenchymal stem cell serum-free complete medium of the present invention (taking adipose stem cells as an example).

[0043] (1) Preparation of adipose stem cells (same as described in most documents): The fat used in the experiment was obtained from abdominal liposuction patients. Under aseptic conditions, 50 mL of a fat-physiological saline mixture was obtained, centrifuged, and washed twice with PBS to remove anesthetics and blood cells to obtain fat particles with higher purity. 0.1% collagenase digestion in a constant temperature shaker at 37°C for 60min, 1500r / min, centrifugation for 10min, remove the upper undigested adipose tissue and grease, resuspend the precipitate and filter on a 100um filter, centrifuge again, red blood cell lysate lysis red blood cells for 5min, phosphate buffer Wash twice and use the following three kinds of culture media:

[0044] 1. Traditional s...

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Abstract

The invention relates to a human mesenchymal stem cell culture medium. According to the culture medium, the basal culture medium comprises the following components based on the final concentration: 1-2g/L of human serum albumin, 5-10mg/L of transferring, 2-8mg/L of fibronectin, 1-4mg/L of laminin, 50g/L of Fe(NO3)3.9H2O, 417g/L of FeSO4.7H2O, 1-3mu g/L of estradiol, 2-5mu g/L of testosterone, 1-3mu g/L of progesterone, 39.25-117.74 mu g/L of dexamethasone, 5-10mg/L of insulin, 376.36mg/L of riboflavin, 80.96-242.87mg/L of coenzyme A, 4.41-6.17mg/L of butanediamine, 1-2mg/L of taurine, 0.61-1.85mg/L of aminoethanol, 8.81-26.42mg/L of pyruvic acid, 3.78-7.56mu g/L of sodium selenate, 292.3-584.6mg/L of L-glutamine, 2-8mu g/L of vascular endothelial growth factor, 4-10mu g/L of epidermal growth factor, 4-10mu g/L of basic fibroblast growth factor, 1-5mu g/L of leukaemia inhibitory factor, 1-5mu g/L of insulin-like growth factor-I and 2-8mu g/L of stem cell factor. The culture medium does not contain the animal serum, the potential animal endogenous endotoxin or virus of the animal serum is eliminated, and the culture medium is conveniently applied to clinics.

Description

Technical field [0001] The invention belongs to the technical field of cell culture, and specifically relates to a human mesenchymal stem cell culture medium. Background technique [0002] Stem cells (SC) are a type of self-renewing pluripotent cells. Under certain conditions, they can differentiate into a variety of functional cells, which can be used to treat leukemia, congenital metabolic diseases, certain Various diseases such as solid tumors, diabetes, heart disease and cerebral palsy have very broad medical uses. As of December 2012, 4178 stem cell clinical experimental studies have been carried out globally, and as many as 602 projects have entered clinical phase III. Mesenchymal stem cells (MSC) are currently the safest, most effective and widely used adult stem cells in treatment, mainly derived from bone marrow, fat, umbilical cord, placenta, amniotic membrane and so on. As of April 2013, 7 kinds of stem cell clinical treatment products have been approved for marketing...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 孙洪喜张高峰于丽范友金陈云燕
Owner QINGDAO RESTORE BIOTECHNOLOGY CO LTD
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