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The extended serial subculture of human diploid fibroblasts on microcarriers using a new medium supplement formulation
Author(s) -
Forestell Sean P.,
Kalogerakis Nicolas,
Behie Leo A.
Publication year - 1992
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260400907
Subject(s) - microcarrier , subculture (biology) , laboratory flask , cell culture , microbiology and biotechnology , cell , biology , chemistry , cell growth , andrology , immunology , biochemistry , medicine , genetics
Abstract Human diploid fibroblasts serially passaged on microcarriers exhibit a decrease in their proliferative capacity with each transfer from microcarrier‐to‐microcarrier. This phenomenon, which does not occur in the same time scale with cells cultured in T‐flasks, has been a serious barrier to the systematic utilization of microcarriers in the scale‐up of anchorage‐dependent human diploid cell cultures. This decreases in cell growth with each passage is shown to be related to the serum content of the medium, with high serum concentrations resulting in a more rapid decrease in cell growth with each serial transfer. As a result, methods for reducing the serum requirement of the cells were investigated. A new medium supplement mixture, PPRF92, has been developed, which allows the serial passaging of MRC5 cells on Cytodex 1 microcarriers through as many as 13 microcarrier‐to‐microcarrier tranfers, and at a serum levels as low as 1%, with no decrease in the proliferative capacity of the cells until they approach their reported population doubling limit. This new supplement mixture is a significant improvement to microcarrier technology in that it enables the use of microcarriers in the early stages of inocculum build‐up for the production purposes. © 1992 John Wiley & Sons, Inc.