Method of harvesting cells on three-dimensional microcarrier

Abstract

The invention discloses a method of harvesting cells on a three-dimensional microcarrier. The method comprises the following steps of 1, executing natural sedimentation or centrifugation of a three-dimensional microcarrier adsorbing cultured cells, and removing the supernatant to obtain a three-dimensional microcarrier containing cells; 2, adding the three-dimensional microcarrier containing the cells into lysate, and incubating and lysing the three-dimensional microcarrier; 3, after the three-dimensional microcarrier in step 2 is completely lysed, a stop buffer is added to terminate the lysisprocess or directly executing the step 4 to terminate the lysis; 4, centrifuging the system obtained in step 3, and discarding the supernatant to obtain the cells on the three-dimensional microcarrier. The cells on the carrier can be more gently obtained for subsequent application; by lysing the microcarrier, the microcarrier is dissolved from a solid state to a liquid state, accordingly the cells on the carrier are released to the solution, and the solution and the cells are separated in a centrifugation way so as to obtain the cells.

Description

technical field [0001] The invention relates to a method for harvesting cells on three-dimensional microcarriers, belonging to the technical field of cell culture. Background technique [0002] With the rapid development of interdisciplinary subjects such as biomedicine, materials science, mechanics, and engineering, more and more people are paying attention to the difference between the microenvironment of cells cultured in vitro and the microenvironment in vivo. The traditional two-dimensional cell culture (based on commercial culture dishes or multi-well plates) technology has been developed for more than 100 years, and it has been widely used in basic life science research, medical research and other fields. However, with the rapid development of microscopic imaging technology, researchers have gradually discovered the living state of cells in a two-dimensional culture environment, and the cell morphology is far from the culture conditions in the real environment in vivo...

AI Technical Summary

Problems solved by technology

However, after three-dimensional cell culture, multi-layered or agglomerated cells are difficult to be digested by trypsin, and the long-term digestion of trypsin or its substitutes will cause great damage to the cells themselves. Therefore, a more gentle method is needed. Methods Harvesting cells from the three-dimensional carrier, that is, by cracking the three-dimensional carrier material without damaging the cells, the efficient harvesting of cells from the three-dimensional carrier material can be achieved, thereby meeting the needs of subsequent applications

For cell culture on 3D microcarriers, there is currently no method for more gentle harvesting of cells on this carrier for subsequent applications

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