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Method used for inducing differentiation of human multipotential stem cells into retinal progenitor cells

A technology of human pluripotent stem cells and retinal cells, applied in the field of biomedicine, can solve the problems of long cell differentiation cycle and low cell differentiation efficiency, and achieve the effect of improving adhesion efficiency and differentiation rate

Inactive Publication Date: 2014-03-12
FUDAN UNIV
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Problems solved by technology

This method can effectively solve the problems caused by defects such as low cell differentiation efficiency and long cell differentiation cycle in retinal cell transplantation therapy

Method used

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  • Method used for inducing differentiation of human multipotential stem cells into retinal progenitor cells
  • Method used for inducing differentiation of human multipotential stem cells into retinal progenitor cells
  • Method used for inducing differentiation of human multipotential stem cells into retinal progenitor cells

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Embodiment 1

[0036] (1) Maintenance and passage of human pluripotent stem cells:

[0037] The culture medium of human pluripotent stem cells was replaced every day, and passaged every 5 to 7 days; before passage, a layer of mouse fibroblast feeder layer should be laid in a 6-well plate to maintain the pluripotent differentiation characteristics of stem cells. When passaging, suck out the culture medium in the well plate, wash it once with DMEM / F12 culture medium, add Dispase (1U / ml) digestion solution 1ml / well after aspirating the culture medium, digest in a 37°C incubator for 2 minutes, and quickly absorb In addition to the digestion solution, add 2ml / well of stem cell culture medium, scrape off the cell clones in the well plate with the tip of a yellow pipette, to ensure that the size of the cell fragments is basically the same, and pipette the cell fragment suspensions of 1 hole and 5 holes into two 15ml In the centrifuge tube, after it settles naturally, suck off the supernatant, add f...

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Abstract

The invention belongs to the field of biomedicine, and specifically relates to a method used for inducing differentiation of human multipotential stem cells into retinal progenitor cells. According to the method, human embryonic stem cells or human induced multipotential stem cells are induced in a medium containing N2, and then are delivered into a retina induction medium containing neural basis culture medium, knockout serum replacement and nicotinamide so as to obtain the retinal progenitor cells. The method is capable of increasing differentiation rate of human multipotential stem cells into retinal progenitor cells; and the retinal progenitor cells obtained via differentiation can be used for transplantation therapy of irreversible degenerative diseases of retinal neurons, such as age-related macular degeneration and retinitis pigmentosa, and also can be used for iPS cells sourced from patients as cell models in drug screening and gene interference experiments. Compared with existing technology, small molecule compounds used in the method are less, so that influences caused by non-anthropogenic factors are avoided effectively, and yield of obtained optic cup-like cells is obviously higher than that of existing induced differentiation methods.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a cell induction method, in particular to a method for inducing human pluripotent stem cells to differentiate into retinal precursor cells. In particular, a method for inducing differentiation of human pluripotent stem cells into retinal precursor cells including neural retinal precursor cells and pigment epithelial precursor cells. Background technique [0002] Among various clinical diseases, central nervous system diseases have always been the focus of attention. There is a consensus in the field that research on embryonic stem cells is of great significance to the treatment of central nervous system diseases. The retina is a part of the central nervous system located in the orbit. In view of its easy access, and with the development of imaging technology and the in-depth understanding of retinal structure and function, people are exploring the process of cell therapy for the treatmen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61K35/44A61P27/02C12Q1/02A61K35/30
Inventor 王晓冰周国民张素春
Owner FUDAN UNIV
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