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A method for promoting the directed differentiation and proliferation of mesenchymal stem cells to neural precursor cells

A technology of neural precursor cells and stem cells, applied in the field of regenerative medicine, to achieve high cell activity, improve neural differentiation efficiency, and promote clustering and proliferation.

Active Publication Date: 2019-11-12
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the movement and proliferation of cells require enough space. The porosity of hydrogels is generally only a dozen microns, which greatly limits the expansion of differentiated neural precursor cells. Therefore, it is necessary to increase the porosity or space inside the hydrogel. Achieving massive proliferation of cells

Method used

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  • A method for promoting the directed differentiation and proliferation of mesenchymal stem cells to neural precursor cells
  • A method for promoting the directed differentiation and proliferation of mesenchymal stem cells to neural precursor cells
  • A method for promoting the directed differentiation and proliferation of mesenchymal stem cells to neural precursor cells

Examples

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Embodiment 1

[0031] Example 1: Promoting directional differentiation and proliferation of human umbilical cord-derived mesenchymal stem cells to neural precursor cells in vitro

[0032]First, prepare arginyl-glycyl-aspartic acid (RGD), isoleucine-lysine-valine-alanine-valine (IKVAV) and tyrosine-isoleucine respectively Sodium alginate modified with amino acid-glycine-serine-arginine (YIGSR). The specific method is to dissolve sodium alginate (molecular weight 500kDa, ratio of guluronic acid to mannuronic acid 2) in 0.1M 2-(N-morpholino)ethanesulfonic acid (MES) buffer containing 0.5M NaCl (pH value 6.5) to obtain a 1% (W / V) sodium alginate solution. Then add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), N-hydroxysulfosuccinimide (sulfo-NHS) and RGD, IKVAV or YIGSR polypeptide, room temperature The reaction was stirred for 24 hours. The molar ratio of EDC to sodium alginate is 1:20, the molar ratio of EDC to sulfo-NHS is 2:1, and the mass ratio of the three polypeptides to sodium ...

Embodiment 2

[0037] Example 2: Promoting the directional differentiation and proliferation of human bone marrow-derived mesenchymal stem cells to neural precursor cells in vitro

[0038] The preparation of sodium alginate modified by RGD, IKVAV and YIGSR was the same as in Example 1. The three kinds of polypeptide-modified sodium alginate powders were dissolved in physiological saline according to the mass ratio of 1:1:1 to obtain a mixed sodium alginate solution with a total concentration of 3% (W / V).

[0039] Thereafter, chondroitin sulfate (average molecular weight: 40 kDa) powder was dissolved in physiological saline to obtain a 10% (W / V) solution. Mix 3% (W / V) modified sodium alginate, 10% (W / V) chondroitin sulfate solution and physiological saline to obtain a final concentration of sodium alginate of 2% (W / V), sodium alginate / chondroitin sulfate A mixed solution with a prime mass ratio of 2.3.

[0040] Then, 10% (W / V) acidic gelatin solution was added dropwise into sterile ethyl ol...

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Abstract

The invention provides a method for promoting directional differentiation and proliferation of mesenchymal stem cells to neural precursor cells. First, the mesenchymal stem cells were embedded in calcium alginate microgel beads containing gelatin microspheres and chondroitin sulfate prepared under the condition of calcium chloride solution, and then the neural induction differentiation medium was added, and in the rotary reactor Culture is carried out to promote the directional differentiation of mesenchymal stem cells to neural precursor cells in vitro and realize their proliferation. After the differentiation is completed, the calcium alginate microgel beads are dissolved with sodium citrate solution, and the differentiated cells are harvested. The invention is simple in operation and low in cost. The gelatin microspheres can provide the necessary growth space for the cells inside the scaffold, the chondroitin sulfate can simulate the biochemical function of the extracellular matrix of the nerve tissue, and the calcium alginate microgel beads provide a structure similar to the extracellular matrix. The three-dimensional structural support and the rotating reactor provide the necessary microgravity environment to further enhance mass transfer and realize its proliferation.

Description

technical field [0001] The invention relates to the field of regenerative medicine, and relates to a method for promoting directional differentiation and proliferation of mesenchymal stem cells to neural precursor cells. Background technique [0002] Neural precursor cells are ideal seed cells for repairing nerve tissue damage due to their ability to differentiate into neurons. However, the source of human neural precursor cells is very difficult, which limits its clinical application. Studies have shown that human mesenchymal stem cells can differentiate into neural precursor cells, which provides a new way for neural tissue repair. However, currently, the induction of mesenchymal stem cells to differentiate into neural precursor cells is carried out under two-dimensional conditions. Studies have found that there is a huge difference between the two-dimensional culture conditions and the three-dimensional growth microenvironment of tissues in vivo, and the cells in two-di...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0797
Inventor 马小军刘洋孙广炜周楠孙东升王淑君廖捍斯肖晶
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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