온도감응성 마이크로캐리어를 이용한 세포 계대배양

Abstract

Microcarrier culture systems are frequently used for large-scale culture of anchorage-dependent mammalian cells. Mass production of a large amount of cells needs serial propagation in microcarrier culture. This requires enzyme treatment which could be a complex step in large-scale cell culture and cause cell damage. Thus, we have developed thermo-sensitive microcarriers by incorporating poly-N-isopropylacrylamide (pNIPAAm) onto Cytodex-3 (R) (pNIPAAm-Cytodex). In this study, we tested the feasibility of serial propagation of Vero cells cultured on pNIPAAm-Cytodex microcarriers in three-dimensional bioreactors. Cells cultured on pNIPAAm-Cytodex were detached from pNIPAAm-Cytodex by a moderate change in temperature without enzyme treatment.. This Would simplify the complex serial propagation steps, avoid cell damage possibly caused by enzyme treatment, and enable one to easily harvest cells, because cells can be detached from microcarriers without enzyme treatment. Vero cells adhered, spread, and grew Successfully on the thermo-sensitive microcarriers similarly to Cytodex microcarriers, and were harvested by simple temperature changes, thereby avoiding the use of proteolytic enzymes and laborious and time-consuming processes. Moreover, Vero cells detached by temperature change reattached and grew on pNIPAAm-Cytodex after scale-Lip in a manner similar to Culture on Cytodex. The pNIPAAm-Cytodex microcarrier Culture method would be useful for the mass production of biologicals through large-scale Suspension Culture of anchorage-dependent mammalian cells.