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Method for inducing human placenta chorionic mesenchymal stem cells to be differentiated into neural stem cells

A kind of stem cell, human placenta technology, applied in animal cells, nervous system cells, vertebrate cells, etc., can solve the problem of nerve cells unable to regenerate, achieve self-renewal and multi-directional differentiation characteristics, easy to induce differentiation and maturation, cells The effect of strong proliferative ability

Inactive Publication Date: 2017-05-10
GUANGZHOU STEMOVE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention provides a method for inducing human placental chorionic mesenchymal stem cells to differentiate into neural-like stem cells, the method does not add chemical induction factors, and solves the problem that existing nerve cells cannot regenerate

Method used

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  • Method for inducing human placenta chorionic mesenchymal stem cells to be differentiated into neural stem cells
  • Method for inducing human placenta chorionic mesenchymal stem cells to be differentiated into neural stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] (1) Obtain mesenchymal stem cells derived from human placental chorion in vitro.

[0025] Take 5 g of placental chorionic tissue and cut it into pieces with sterile surgical scissors or a knife to obtain tissue pieces with a side length of less than 3 mm. Collect the obtained tissue pieces in a 15ml centrifuge tube, add an equal volume of phosphate buffer, shake vigorously and let it stand at room temperature for 5 minutes. After the tissue pieces are divided into two layers, carefully collect the upper layer. The collected upper layer tissue pieces use phosphate Wash with buffer three times; add an equal volume of 0.075% collagenase I to the collected upper tissue block, and bathe in water at 37°C for 30 minutes; add an equal volume of DMEM medium containing 10% fetal bovine serum to stop digestion, and mix thoroughly Let stand at room temperature for 10 minutes to separate layers; remove the upper layer (lipid / debris), centrifuge at 20°C and 280g for 5 minutes to coll...

Embodiment 2

[0032] (1) Obtaining mesenchymal stem cells derived from human placental chorion in vitro: Same as Example 1.

[0033] (2) Induce the differentiation of human placental chorionic mesenchymal stem cells into neural-like stem cells.

[0034] Polymerization of three-dimensional scaffolds: Add 2.5 mg / mL polylactic acid solution to a 12-well culture plate containing three-dimensional materials, 25 μL / well, and polymerize in a 37°C incubator for 30 minutes to obtain a three-dimensional porous polymerized gel. stand.

[0035] Add the mixed human placental chorionic mesenchymal stem cell suspension to the three-dimensional scaffold that has been polymerized to make the cell density 5×10 5 cells / hole. Add 500ul of human placental chorionic mesenchymal stem cell culture medium (L-DMEM+10%FBS+100u / L penicillin+100u / L streptomycin), place in an incubator at 37°C, 5%CO 2 Culture was carried out, and the culture medium was replaced every other day.

[0036] (3) Obtain neural-like stem c...

Embodiment 3

[0038] (1) Obtaining mesenchymal stem cells derived from human placental chorion in vitro: Same as Example 1.

[0039] (2) Induce the differentiation of human placental chorionic mesenchymal stem cells into neural-like stem cells.

[0040] Polymerization of three-dimensional scaffolds: Add 5 mg / mL silk fibroin solution to a 12-well culture plate containing three-dimensional materials, 25 μL / well, and polymerize in a 37°C incubator for 30 minutes to obtain a porous polymerized gel. 3D bracket.

[0041] Add the mixed human placental chorionic mesenchymal stem cell suspension to the three-dimensional scaffold that has been polymerized to make the cell density 5×10 5 cells / hole. Add 500ul of human placental chorionic mesenchymal stem cell culture medium (L-DMEM+10%FBS+100u / L penicillin+100u / L streptomycin), place in an incubator at 37°C, 5%CO 2 Culture was carried out, and the culture medium was changed every other day.

[0042] (3) Obtain neural-like stem cells.

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Abstract

The invention discloses a method for inducing human placenta chorionic mesenchymal stem cells to be differentiated into neural stem cells through three-dimensional culture. The method comprises the following steps of mixing three-dimensional materials with a coagel solution; performing coagel treatment for 30 minutes at 37 DEG C; then, adding the human placenta chorionic mesenchymal stem cells; performing culture at 37 DEG C by 5-percent CO<2>; and replacing the culture solution every other day, wherein the coagel solution is a type I collagen solution, a polylactic acid solution or a silk fibroin solution. An in-vivo three-dimensional growth environment is provided for the neural differentiation by using a three-dimensional culture technology; the relation among cells is promoted, so that the cells from multilayer growth; meanwhile, the self updating and multi-directional differentiation features of the stem cells can be better maintained; no chemical inducing factor effect exists, so that the differentiated neural stem cells can be directly used for subsequent clinic tests; and the safety is higher.

Description

technical field [0001] The invention relates to a method for inducing human placental chorionic mesenchymal stem cells to differentiate into neural-like stem cells, in particular to a method for differentiating human placental chorionic mesenchymal stem cells into neural-like stem cells in a three-dimensional culture method without chemical induction factors. Background technique [0002] Neural stem cells (neural stem cells, NSCs) are a kind of mother cells with division potential and self-renewal ability, which can produce various types of cells of nerve tissue through asymmetric division. For a long time, it has been believed that the nerve cells in the adult mammalian brain do not have the ability to regenerate, and cannot regenerate once damaged or even dead. This point of view has greatly limited the treatment of central nervous system diseases. Although traditional medicine, surgery and rehabilitation have made some progress, they still cannot achieve satisfactory re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797C12N5/0775
Inventor 冯文峰黄鹏昌芒
Owner GUANGZHOU STEMOVE BIOTECH CO LTD
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