Culture medium, induction culture method of oligodendrocyte progenitor cells and application

Abstract

The invention provides a culture medium, an induction culture method of oligodendrocyte progenitor cells and application. The culture medium combination provided by the invention comprises the following culture medium in the induced differentiation stage: embryonic bodies, neural epithelial cells, Pre-OPCs and hOPCs. In each stage, a specific induction culture medium is adopted for adherent culture. Experimental results show that the induction rate of differentiation of hESC and hiPSC to hOPC can be remarkably increased, the purity of hOPC is obviously improved, and the induction time is greatly shortened compared with that of traditional suspension culture. The induction culture method is suitable for differentiation of human embryonic stem cells and human induced pluripotent stem cells to oligodendrocyte progenitor cells. Experiments show that OPC obtained through induction culture can obviously reduce the loss of injured brain region tissues, can improve and enhance the recovery of the injured brain region tissues, and can be used for cell transplantation to treat cerebral nervous system diseases such as cerebral apoplexy injury.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a culture medium and a method for inducing and culturing oligodendrocyte progenitor cells and its application. Background technique [0002] Stroke is the leading cause of death and disability in the world. At present, there is a lack of effective treatment methods at home and abroad, especially in the recovery stage. White matter damage is an important pathological change in most strokes, manifested as oligodendrocyte (OL) damage, nerve demyelination, and axon loss. During stroke recovery, the brain does not provide sufficient oligodendrocyte progenitor cells (OPCs) to generate OL and form myelin. Therefore, providing exogenous OPCs to generate new OL and nerve myelin for the recovery of axonal structure and function can be a new hope for the treatment of ischemic stroke. We previously used hiPSC-induced OPCs to treat demyelinating lesions in rodents with remarkable effi...

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Problems solved by technology

The disadvantage of this process is that (1) it is cumbersome, time-consuming, difficult to replace the cell culture medium, and the living cells are unstable and difficult to quality control; (2) the cell pellets need to be cut into small cell clumps with a blade, The process is more cumbersome and complicated, and it is not easy to ensure quality control and aseptic operation, and it is difficult to turn it into large-scale production; (3) In the three-dimensional suspension culture stage, the growth status of cells cannot be tracked and tested by immunolabeling or other forms; (4) The state of cell pellets cannot be directly cryopreserved, subcultured, and expanded for culture, so that each induction culture must start from the state of NE cells, prolonging the experimental time and increasing the workload; (5) Finally, during the culture process of hOPCs, use During the process of cutting the cell ball with a sterile blade, it is easy to produce plastic fragments mixed in the cells, which is not conducive to the subsequent transplantation experiments or cell purification experiments, and is especially not conducive to the large-scale industrial production of hOPC in the future

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