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Development of oligodendrocyte progenitor cells prepared by differentiating mesenchymal stem cells through eleutheroside and kit, and medicine for nerve system disease

A technology of precursor cells and stem cells, which is used in nervous system diseases, nervous system cells, medical preparations containing active ingredients, etc. Complex and other problems, to achieve the effect of improved safety, strong selectivity and good safety

Inactive Publication Date: 2012-02-01
北京弘润天源基因生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to overcome the existing methods for inducing and differentiating human mesenchymal stem cells into oligodendrocytes with complicated operation, high cost, low purity of differentiated oligodendrocytes, and the induction method used in the separation and expansion process. The drug contains harmful drugs to the human body, which makes the oligodendrocytes obtained by induction and differentiation poor in safety when prepared into a pharmaceutical composition, and provides a differentiation medium for inducing differentiation of human mesenchymal stem cells into oligodendrocytes , the method of separating and expanding oligodendrocytes using this medium is simple to operate, low in cost, high in differentiation purity, and stable in nature

Method used

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  • Development of oligodendrocyte progenitor cells prepared by differentiating mesenchymal stem cells through eleutheroside and kit, and medicine for nerve system disease
  • Development of oligodendrocyte progenitor cells prepared by differentiating mesenchymal stem cells through eleutheroside and kit, and medicine for nerve system disease
  • Development of oligodendrocyte progenitor cells prepared by differentiating mesenchymal stem cells through eleutheroside and kit, and medicine for nerve system disease

Examples

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Effect test

Embodiment 1

[0040] This example is used to illustrate the method of using the differentiation medium of the present invention to differentiate human bone marrow mesenchymal stem cells into oligodendrocyte precursor cells.

[0041] Human bone marrow was obtained from a 31-year-old donor, and an informed consent was signed with the donor and the hospital. The human bone marrow was diluted with 1×PBS equal volume to obtain the human bone marrow dilution, and the volume ratio of human bone marrow dilution: 1.077 kg / m3 Ficoll-Hypaque (Ficoll-Hypaque) was added to the In a 50ml centrifuge tube, perform density gradient centrifugation at 1500rpm, 25 degrees for 10 minutes, collect the middle layer cells after centrifugation, and use 1×PBS 1000rpm centrifugation for 10 minutes, wash the obtained cells three times, add complete medium containing 10% fetal bovine α-MEM medium (Gibco, USA) of serum (Hyclone, USA). After cell counting, press 3×10 5 cells / cm 2 The density is inoculated at 75cm 2 A...

Embodiment 2

[0045] This example is used to illustrate the method of differentiating umbilical cord-derived mesenchymal stem cells into oligodendrocyte precursor cells using the differentiation medium of the present invention.

[0046] The umbilical cords were collected from 26-year-old healthy pregnant women after delivery. An informed consent was signed with the pregnant woman and the hospital to collect her umbilical cord after delivery.

[0047]Wash the isolated tissue umbilical cord three times with 1×PBS containing 2% double antibodies (penicillin and streptomycin), and then cut it into pieces of about 1mm with ophthalmic scissors 3 of small pieces. The resulting shredded tissue pieces were digested with 0.25% trypsin solution at a weight-to-volume ratio of 1 g: 0.5 mL for 5 minutes, then added fetal bovine serum that was one-fifth of the volume of trypsin solution to terminate the digestion, and centrifuged at 1000 rpm for 5 minutes Collect the digested tissue pieces, pour off the...

Embodiment 3

[0052] This example is used to illustrate the method of using the differentiation medium of the present invention to subculture and expand culture of oligodendrocyte precursor cells in vitro, and the method of cryopreservation of the obtained oligodendrocyte precursor cells.

[0053] Take the oligodendrocyte precursor cells obtained in Examples 1 and 2, when the adherent growth reaches 90% of the bottom surface of the culture bottle, the cells are digested with trypsin, centrifuged at 800rpm for 5 minutes, and then collected in 50ug / mL eleutheroside, 10% fetal bovine serum, 20ng / mL EGF and 20ng / mL bFGF α-MEM medium for subculture. Similarly, when the adherent growth of the oligodendrocyte precursor cells reached 90% of the bottom surface of the culture bottle, the above subculture process was repeated. When the oligodendrocyte precursor cells were amplified to the 5th passage, a bottle of oligodendrocyte precursor cells was digested with trypsin, centrifuged at 800rpm for 5 m...

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Abstract

The invention provides a differential medium for obtaining oligodendrocyte progenitor cells and proliferation thereof by processing mesenchymal stem cells through eleutheroside. The differential medium comprises a liquid basal medium, and also comprises 1-100ug / mL eleutheroside, 1 to 20 percent of fetal calf serum, 5 to 300ng / mL human epidermal growth factor (EGF) and 5 to 300ng / mL basic fibroblast growth factor (bFGF) based on the volume of the liquid basal medium. The invention also provides a method for preparing an eleutheroside processing kit by using the differential medium, and also provides a medicinal composition comprising the oligodendrocyte progenitor cells prepared by the method and used for treating nerve system disease. By using the differential medium, the method has the advantages that: the mesenchymal stem cells can be efficiently differentiated and the oligodendrocyte progenitor cells are obtained. The medicinal composition for treating diseases such as traumatic spinal cord injury, multiple sclerosis and the like can repair injured nerve cells in vivo and eliminate neurologic disorder.

Description

technical field [0001] The present invention relates to a kind of human mesenchymal stem cells treated with eleutheroside to obtain oligodendrocyte precursor cells (Oligodendrocyte Progenitor cells, OPC) and differentiation medium for proliferation thereof, and a method for developing a kit thereof, as well as obtaining A pharmaceutical composition of precursor cells for treating traumatic spinal cord injury, multiple sclerosis and other diseases. More specifically, the present invention relates to a differentiation medium for treating human mesenchymal stem cells with eleutheroside to obtain oligodendrocyte precursor cells and their proliferation, using the medium to selectively obtain oligodendrocyte precursor cells method, and a pharmaceutical composition comprising the oligodendrocyte precursor cells obtained by the method for treating traumatic spinal cord injury, multiple sclerosis and other diseases. Background technique [0002] Oligodendrocytes are a type of macrog...

Claims

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Application Information

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IPC IPC(8): C12N5/0797A61K35/14A61K35/24A61K35/44A61P25/00A61P25/28A61K35/30
Inventor 沈丽黄启明郝丽敏王振光
Owner 北京弘润天源基因生物技术有限公司
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