Unlock instant, AI-driven research and patent intelligence for your innovation.

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, great stability effect

Active Publication Date: 2015-04-29
QINGDAO RESTORE BIOTECHNOLOGY CO LTD
View PDF6 Cites 0 Cited by
  • 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 for a long time
3. Without a full set of third-party pathogen testing (14 tests), a full set of third-party pathogen testing reports cannot be provided. The cultured cells have the risk of pathogen contamination and have not reached the clinical level.
4. Several serum-free culture media for human mesenchymal stem cells can be purchased in the market, but all of them are imported from abroad, and the price is expensive. The price per 500ml is more than 3,000 yuan, but the culture effect is not ideal. The expression of stem cell surface markers is not ideal
And it is for research application only, not for human diagnosis and treatment

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 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 serum-free complete medium for clinical-grade human mesenchymal stem cells provided by the present invention. The composition and concentration are shown in Table 1. The following components are dissolved according to their respective characteristics, mixed, and HEPES is constant volume, 0.22 μm Sterilize by filtration and store at 4°C.

[0035] Table 1 The 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 are up to standard, as shown in Table 2:

[0039] Table 2 Third-party quality inspection items and results of 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 serum-free complete culture medium for clinical grade human mesenchymal stem cells of the present invention is qualified in all detection indexes. The culture medium that has passed the test can only be used clinically.

Embodiment 3

[0042] Embodiment 3. The 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-derived stem cells as an example).

[0043] (1) Preparation of adipose-derived stem cells (same as described in most literatures): The fat used in the experiment was obtained from patients undergoing abdominal liposuction. Under aseptic conditions, 50 mL of fat saline mixture was obtained, centrifuged, washed twice with PBS to remove anesthetics and blood cells, and fat particles with high purity were obtained. Digest with 0.1% collagenase at 37°C for 60 minutes, centrifuge at 1500r / min for 10 minutes, remove the undigested fat tissue and oil in the upper layer, resuspend the sediment with a 100um filter, and centrifuge again. Wash twice, and culture in the following three media respectively:

[0044] 1. Traditional serum-containing medium (DMEM / F12+10%FBS+8ng / mlbFGF);

[0045] 2...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

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 in particular relates to a culture medium for human mesenchymal stem cells. Background technique [0002] Stem cells (SC) are a kind of pluripotent cells with self-renewing ability. 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 applications. As of December 2012, 4,178 stem cell clinical trials have been carried out around the world, and as many as 602 projects have entered the third phase of clinical trials. 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, etc. As of April 2013, 7 stem cell clinical treatment products hav...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775
Inventor 刘海英王娟陈立材张炳强王清华许评
Owner QINGDAO RESTORE BIOTECHNOLOGY CO LTD