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A method for inducing differentiation of umbilical cord mesenchymal stem cells into dopaminergic neurons

A technology for inducing differentiation of MSCs, applied in the field of neurobiology, can solve the problems of directional induction and differentiation of MSCs that are not disclosed, and achieve the effect of improving efficiency and selectivity

Inactive Publication Date: 2017-10-27
THE THIRD MEDICAL CENT OF THE CHINESE PEOPLES LIBERATION ARMY GENERAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this patent document does not disclose how to effectively induce MSCs to differentiate into dopaminergic neurons, and there are very few studies on how to effectively induce MSCs to differentiate into dopaminergic neurons in other prior art

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Acquisition and culture of umbilical cord mesenchymal stem cells

[0043] Neonatal umbilical cords authorized with maternal consent were used as a source of mesenchymal stem cells.

[0044] The neonatal umbilical cord was washed three times in physiological saline containing 1% double antibody to remove the blood stains on the surface, and then cut into small pieces about 1 cm long. Use ophthalmic scissors to cut the umbilical cord longitudinally along the direction parallel to the blood vessels, and peel off the 2 umbilical arteries and 1 umbilical vein from the umbilical cord. Peel off the amniotic membrane on the surface, wash the Huatong glue part with normal saline containing 1% double antibody for 3 times, and cut it into pieces to about 1mm 3 size. Evenly spread the shredded tissue blocks on the 75cm 2 Place in the culture bottle at room temperature for 5-10 minutes to make the tissue pieces stick tightly. Add 5ml of medium containing DMEM / F12, 2% ...

Embodiment 2

[0046] Example 2: Directed differentiation of umbilical cord mesenchymal stem cells in vitro

[0047] Freshly passaged P3-6 umbilical cord mesenchymal stem cells were used in 5×10 4 Inoculate in a 24-well plate with poly-lysine-coated coverslips per ml, 1ml / well, replace the pre-induction culture medium after 3 days of adherent culture, change the medium in half after 3 days, and terminate the induction on the 3rd day .

[0048] The pre-induction culture medium is DMEM as the base medium, which also contains 50nmol vitamin C, 20U / ml human leukocyte inhibitory factor, 8mM L-glutamine, 5mM thioglycerol (MTG), 80U / ml penicillin, 80μg / ml streptomycin.

[0049] The cells obtained from the pre-induction culture were divided into 5×10 4 / ml cell density inoculation for single-layer adherent induction culture, after 2 days of adherent culture, replace the induction medium for induction culture, change the medium after 3 days, stop the induction on the 3rd day, add no inducer to th...

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Abstract

Disclosed is a method for inducing differentiation of umbilical cord mesenchymal stem cells into dopaminergic neurons. The method adopts a two-step combination of pre-induction and induction, and prepares optimized pre-induction medium and induction medium, in which umbilical cord mesenchymal stem cells are successfully induced to differentiate into dopaminergic cells by adding structurally improved glycoside compounds as inducers. Nervous, and a higher proportion of differentiation.

Description

technical field [0001] The invention relates to the field of neurobiology, in particular to a method for inducing differentiation of umbilical cord mesenchymal stem cells into dopaminergic neurons, and the invention also relates to cultured neurons. Background technique [0002] Since Erices et al. reported in 2000 that umbilical cord blood is rich in hematopoietic stem cells and mesenchymal stem cells, umbilical cord blood mesenchymal stem cells have attracted the attention of researchers due to their multi-directional differentiation potential, immunoregulatory effect and no ethical issues. . Subsequently, some scholars reported that umbilical cord blood mesenchymal stem cells could induce the expression of neuronal markers and glial cell markers in vitro. At present, people have been able to successfully isolate and culture umbilical cord blood mesenchymal stem cells in vitro, and have a further understanding of their biological characteristics, surface markers and immun...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0793C12N5/0775
Inventor 安沂华董健伸
Owner THE THIRD MEDICAL CENT OF THE CHINESE PEOPLES LIBERATION ARMY GENERAL HOSPITAL
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