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Method for promoting directional differentiation and proliferation of mesenchymal stem cells towards neural precursor cells

A technology of neural precursor cells and stem cells, which is applied in the field of regenerative medicine, achieves the effects of simple method, improved neural differentiation efficiency, and avoidance of expensive materials and processes

Active Publication Date: 2016-07-06
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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  • Method for promoting directional differentiation and proliferation of mesenchymal stem cells towards neural precursor cells
  • Method for promoting directional differentiation and proliferation of mesenchymal stem cells towards neural precursor cells
  • Method for promoting directional differentiation and proliferation of mesenchymal stem cells towards neural precursor cells

Examples

Experimental program
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Effect test

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, guluronic acid and mannuronic acid ratio 2) in 0.1M 2-(N-morpholino)ethanesulfonic acid (MES) buffer containing 0.5M NaCl ( pH value 6.5), to obtain 1% (W / V) (unit: g / mL) 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...

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) (unit: g / mL).

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

[0040] Then, 10% (W / V) (unit: g / mL) ac...

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Abstract

The invention provides a method for promoting directional differentiation and proliferation of mesenchymal stem cells towards neural precursor cells. The method comprises the following steps: firstly, embedding the mesenchymal stem cells in calcium alginate microbeads which are prepared with the presence of a calcium chloride solution and contains gelatin microspheres and chondroitin sulfate; then, adding a neural induction differentiation medium, and cultivating in a rotary reactor, so as to promote the in vitro directional differentiation and proliferation of the mesenchymal stem cells towards the neural precursor cells; and after differentiation, dissolving the calcium alginate microbeads by virtue of a sodium citrate solution, so that differentiated cells are obtained. The method disclosed by the invention is simple to operate and low in cost; by virtue of the gelatin microspheres, a necessary growth space is provided for internal cells of a stent; by virtue of the chondroitin, the biochemical functions of a neural tissue extracellular matrix can be simulated; by virtue of the calcium alginate microbeads, three-dimensional support, which is similar to the extracellular matrix, is provided; and by virtue of the rotary reactor, a necessary micro-gravity environment is provided, so that mass transfer is further enhanced and proliferation of the cells is achieved.

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 Applications(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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