Method for inductively differentiating umbilical cord mesenchymal stem cells into oligodendroglia cells

A technology of oligodendrocytes and mesenchymal stem cells, applied in the field of neurobiology, can solve the problems of undisclosed MSC directional induction and differentiation, and achieve the effect of improving efficiency and selectivity

Inactive Publication Date: 2015-07-15
中国人民解放军总医院第三医学中心
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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 oligodendrocytes, and there are very few studies on how to effectively induce MSCs to differentiate into oligodendrocytes in other prior art

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  • Method for inductively differentiating umbilical cord mesenchymal stem cells into oligodendroglia cells

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Embodiment 1

[0044] Example 1: Acquisition and culture of umbilical cord mesenchymal stem cells

[0045] Neonatal umbilical cords authorized with maternal consent were used as a source of mesenchymal stem cells.

[0046] 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

[0048] Example 2: Directed differentiation of umbilical cord mesenchymal stem cells in vitro

[0049] 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 .

[0050] The pre-induction medium is DMEM as the basal medium, which also contains 50nmol vitamin C, 20U / ml human leukocyte inhibitory factor, 20μg / L bFGF, 15μg / L EGF, 8mM L-glutamine, 5mM thioglycerol (MTG), 80 U / ml penicillin, 80 μg / ml streptomycin, and the pre-induction medium additionally contains 0.01 M phosphate for buffering.

[0051] The cells obtained from the pre-induction culture were divided into 5×10 4 Inoculate at a cell density of / ml for single-layer adherent induction culture. After 2 days of adherent culture, replace the induction medium ...

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Abstract

The invention discloses a method for inductively differentiating umbilical cord mesenchymal stem cells into oligodendroglia cells. A pre-inducing and inducing two-step combination method is adopted in the method, an optimized pre-inducing culture solution and an optimized inducing culture solution are prepared, saponins compounds are added to serve as an inducing object, and therefore the umbilical cord mesenchymal stem cells are successfully induced to be differentiated into the oligodendroglia cells; in addition, the differentiating proportion is higher.

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 oligodendrocytes, and the invention also relates to cultured oligodendrocytes. 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, glial cell markers, and oligodendrocytes 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, s...

Claims

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Application Information

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IPC IPC(8): C12N5/079C12N5/0775
Inventor 安沂华董健伸
Owner 中国人民解放军总医院第三医学中心
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