Feeder-layer-free culture method for cynomolgus monkey stem cells

Abstract

The invention belongs to the technical field of biology and new medicine, and discloses a feed-layer-free culture method for cynomolgus monkey stem cells. The feed-layer-free culture method comprises the following steps: digesting pluripotent stem cells cultured in a feed layer with collagenase, respinning the digested cells with a feed-layer-free culture medium, transferring the cells into a culture dish coated with matrigel in advance, and putting the culture dish into an incubator; and when the cell density is 80%-90% of the area of the culture dish, digesting the cells with ACCTUSE enzyme, and transferring the cells into the culture dish coated in advance according to the ratio of (1: 5)-(1: 10). According to the method, new culture conditions are developed, so that the cynomolgus monkey pluripotent stem cells can be cultured under the condition of no feed layer, a lot of manpower and material resources are saved without preparation of the feed layer, culture components are definite, and the pluripotency of the cynomolgus monkey pluripotent stem cells can be kept for a long time. The feed-layer-free culture method for cynomolgus monkey stem cells is free of influence of batch detection, the accuracy of experiments can be improved, and a culture system can be clinically used for reference.

Description

technical field [0001] The invention belongs to the technical field of biology and new medicine, and in particular relates to a feeder-free culture method for cynomolgus monkey stem cells. Background technique [0002] At present, the culture of pluripotent stem cells for cultivating cynomolgus monkeys requires a feeder layer to provide some nutrients. The feeder layer is made of 12.5-day-old mouse embryos. After the viscera, limbs and head are removed, the remaining tissue is digested into a single cell culture, and the cells can be used as a feeder layer after the cells proliferate. Therefore, the preparation of the feeder layer needs to spend a lot of manpower and material resources, and because the feeder layer contains animal-derived components, the composition of the medium is uncertain, and the situation of unstable performance is likely to occur, resulting in inconsistent results between batches before and after, and the quality of the medium is unstable and inconsis...

AI Technical Summary

Problems solved by technology

But so far, the culture system for cynomolgus monkey feeder-free stem cell culture has not been well developed. Therefore, a new feeder-free culture method for cynomolgus monkey stem cells is urgently needed

[0003] Through the above analysis, the problems and defects in the prior art are: the preparation of the existing feeder layer requires a lot of manpower and material resources, and the feeder layer contains animal-derived components, and the composition of the medium is uncertain, which is prone to unstable performance. Inconsistencies in the batch results before and after, the unstable and non-standard quality of the culture medium, directly affect the accuracy of the research results

[0004] The difficulty of solving the above problems and defects is: exploring a new training method requires more manpower and material resources, and requires a certain reserve of professional knowledge

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