Premium
Establishment of Higher Passage PER.C6 Cells for Adenovirus Manufacture
Author(s) -
Berdichevsky Marina,
Gentile MariePierre,
Hughes Benjamin,
Meis Peter,
Peltier Joseph,
Blumentals Ilse,
Auniņš John,
Altaras Nedim Emil
Publication year - 2008
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp070258u
Subject(s) - cell , cell growth , doubling time , population , biology , cell culture , cell division , inoculation , immunology , biochemistry , genetics , medicine , environmental health
PER.C6 cells, an industrially relevant cell line for adenovirus manufacture, were extensively passaged in serum‐free suspension cell culture to better adapt them to process conditions. The changes in cell physiology that occurred during this passaging were characterized by investigating cell growth, cell size, metabolism, and cultivation of replication‐deficient adenovirus. The changes in cell physiology occurred gradually as the population doubling level, the number of times the cell population had doubled, increased. Higher passage PER.C6 (HP PER.C6) proliferated at a specific growth rate of 0.043 h ‐1 , 2‐fold faster than lower passage PER.C6, and were capable of proliferation from lower inoculation cell densities. HP PER.C6 cell volume was 16% greater, and cellular yields on glucose, lactate, oxygen, and amino acids were greater as well. In batch cultures, HP PER.C6 cells volumetrically produced 3‐fold more adenovirus, confirmed with three different constructs. The increase in productivity was also seen on a cell‐specific basis. Although HP PER.C6 were more sensitive to the “cell density effect”, requiring lower infection cell densities for optimal specific productivity, they proliferated more after infection than lower passage PER.C6, increasing the number of cells available for virus production. The extensive passaging established HP PER.C6 cells with several desirable attributes for adenovirus manufacture.