Method for 3D culture of autologous fat MSCs derived neural precursor cells

Abstract

The invention discloses a method for 3D culture of autologous fat MSCs (mesenchymal stem cells) derived neural precursor cells. The method includes the steps of: S1. preparing a collagen 3D scaffold; S2. collecting adipose tissue and separating adipose cells, using an MSCs medium for culture to harvest P0 generation MSCs, resuspending the P0 generation MSCs with the MSCs medium, and performing culture to harvest P1 generation MSCs; S3. pretreating the collagen 3D scaffold; S4. using the MSCs medium to resuspend the P1 generation MSCs, conducting inoculation into a tissue culture dish containing the collagen 3D scaffold, and using an NPCs (neural precursor cells) medium to conduct further culture for 6 days, thus harvesting NPCs. The method provided by the invention employs laminin and vitronectin to coat collagen sponge so as to construct the collagen 3D scaffold, can promote MSCs adhesion and inhibit neuronal differentiation, then uses the MSCs medium for pretreatment of the collagen 3D scaffold so as to prevent agglomeration of MSCs during adhesion, and maximumly guarantees single cell adhesion. According to the invention, abdominal adipose tissue is collected and separated, MSCs is amplified, and the collagen 3D scaffold is employed to culture NPCs, and the obtained NPCs can meet the purity and preparation demands of autologous fat MSCs derived NPCs.

Description

A 3D method for culturing neural progenitor cells derived from autologous adipose MSCs technical field The invention relates to the technical field of cell culture, in particular to a method for 3D culturing neural precursor cells derived from autologous adipose MSCs. Background technique Neural stem cells and neural progenitor cells are the precursor cells of nerve cells such as neurons and glial cells, and there is no accurate method to distinguish between them. Neural precursor cells (neural precursor cells, NPCs) play an important role in the development of the nervous system and the repair of nerve damage. The difficulty in repairing adult nerve damage is related to the lack or silence of NPCs. Previous studies have shown that transplantation of allogeneic NPCs can alleviate Parkinson's disease, multiple sclerosis, glioma and other diseases, can promote neurodegenerative diseases, trauma and other axonal injury remyelination and repair, and relieve clinical symptoms. ...

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Problems solved by technology

Neurosphere culture is to inoculate MSCs into hydrophobic culture containers to form multi-cell aggregated cell spheres. It belongs to the semi-suspension culture system. Its culture capa...

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