Neonatal pig islet cell culture medium and application method thereof

Abstract

The invention discloses a neonatal pig islet cell culture medium and an application method thereof. The method for preparing the neonatal pig islet culture medium includes adding 10 micromoles to 50 micromoles of a cell apoptotic inhibitor Z-VAD-FMK, human albumin accounting for 15% to 25% of the culture medium and pig serum accounting for 5% to 15% of the culture medium to an HAM'S / F-10 culture medium, and adding 6 micromoles to 10 micromoles of sodium pyruvate ro each liter of the culture medium. The neonatal pig islet culture medium is an improved neonatal pig islet culture medium system which can provide a large number of active and stable yields.

Description

technical field [0001] The invention relates to a newborn pig islet cell culture medium and a using method thereof. Background technique [0002] The current porcine islet cell culture system is to remove the pancreas from the animal body to separate the islet cells, and culture them in CMRL1066, RPMI and other media to maintain their growth or induce their directional differentiation. The cultured islet cells are generally used for transplantation. Surgery for diabetes. Nowadays, there are many methods for culturing neonatal pig islet cells. Generally, neonatal pigs that are 1-5 days old are used to dissect the pancreas, which is broken and digested with collagenase to form a cell mass, which is placed in a cell culture medium for 3-5 days. The amount of active and functional islet cells varies (2000IEQ / g-4000IEQ / g), and the purity of cultured islet cells also varies (70-90%). Islet cells will be lost due to apoptosis even in low temperature culture, especially β cells, a...

AI Technical Summary

Problems solved by technology

Islet cells will be lost due to apoptosis even in low temperature culture, especially β cells, and the length of the culture process is difficult to choose. If the culture time is short, the β cells will not mature, and if the culture time is too long, a large number of islet cells will be lost due to Apoptosis is lost, the general recovery rate is only about 60%, and the same amount of islet cell insulin secretion is reduced due to β cell apoptosis (insulin / DNA value is 49.00±1.95mIU / g)

It is difficult to remove contamination during the culture process, and it is easy to bring inflammatory reactions to the subsequent transplantation process

At present, there is no islet cell culture method that can provide high-purity, active and stable yield in large quantities, so how to establish an optimized medium composition and culture method has become the most urgent issue for islet cell culture