Perfusion culture apparatus for suspended mammalian cells

Abstract

A variety of processes have been proposed for mammalian cell culture in the commercial production of useful substances (e.g., monoclonal antibodies, therapeutic and diagnostic proteins). Among them, the perfusion culture of suspended non-immobilized cells is the most advantageous. Perfusion culture can be classified by the separation process of suspended cells from the culture mixture into three types, namely filtration, gravitational settling and centrifugation. From a commercial point of view, the present situation and technical problems of suspended cell perfusion culture will be reviewed based on the three types. The recent development of perfusion culture has been carried out mainly on the filtration separation process, but the centrifugation process seems to have a promising future because of operation stability and scale-up feasibility. The reasons will be explained in details.

Apoptosis-resistant E1B-19K-expressing NS/0 myeloma cells exhibit increased viability and chimeric antibody productivity under perfusion culture conditions

“…This has been observed for cells cultivated in batch (Ramirez and Mutharasan, 1990), fed-batch (de Tremblay et al, 1993;Robinson et al, 1994), chemostat (Boraston et al, 1984), and perfusion modes (Al-Rubeai et al, 1992;Banik and Heath, 1995;Batt et al, 1990;de la Broise et al, 1991;Johnson et al, 1996;Hansen et al, 1993;Park and Ryu, 1994;Tokashiki and Takamatsu, 1993;Trampler et al, 1994), as well as in cells treated with cell-cycle blocking agents (Al-Rubeai and Emery, 1990;Jayme, 1991;Mercille and Massie, 1998;Oh et al, 1995;Øyaas et al, 1994a,b;Reddy and Miller, 1994;Suzuki and Ollis, 1990;Takahachi et al, 1994). Indeed, specific antibody production has been shown to be cell cycle dependent and optimum in the G 1 and/or G 2 /M phase (Cazzador and Mariani, 1993;Kromenaker and Srienc, 1991;Linardos et al, 1992;Richieri et al, 1991;Suzuki and Ollis, 1989).…”

“…The significant reduction in the growth rate of E1B-19K-expressing cells was also reflected in an increase in the perfusion to batch q MAb ratio. Indeed, many mammalian cell lines cultivated in perfusion exhibit an increase in specific MAb productivity under such slow growth conditions (Al-Rubeai et al, 1992;Banik and Heath, 1995;Batt et al, 1990;de la Broise et al, 1991de la Broise et al, , 1992Johnson et al, 1996;Hansen et al, 1993;Mercille et al, 1994a-c;Shi et al, 1993;Tokashiki and Takamatsu, 1993;Trampler et al, 1994). The q MAb has been shown to be cell cycle dependent and optimum in the G 1 (Byars and Kidson, 1970;Cazzador and Mariani, 1993;Garatun-Tjeldsto et al, 1976;Linardos et al, 1992;Mercille and Massie, 1998;Ramirez and Mutharasan, 1990;Richieri et al, 1991;Suzuki and Ollis, 1989) or G 2 /M phase (Cherlet et al, 1995;Kromenaker and Srienc, 1991;Mercille and Massie, 1998;Watanabe et al, 1973).…”

“…Perfusion presents a number of advantages over other modes of cultivation including increased volumetric productivity, reduced labor and costs, and rapid removal of easily inactivated products from the culture environment (Prior et al, 1989). However, although high density cultures of hybridoma or recombinant myeloma cells can be achieved, decreasing viabilities are observed in a number of different perfusion systems (Al-Rubeai et al, 1992;Avgerinos et al, 1990;Banik and Heath, 1994;de la Broise et al, 1991de la Broise et al, , 1992Hansen et al, 1993;Hiller et al, 1993;Johnson et al, 1996;Kitano et al, 1986;Mercille et al, 1994c;Reuveny et al, 1986;Schmid et al, 1992;Tokashiki and Takamatsu, 1993;Trampler et al, 1994;Van Erp et al, 1991). Under these conditions, an equilibrium is established in which a significant loss of cell viability is matched by a high level of proliferation, leading to a significant waste of metabolic energy that could be more appropriately diverted to recombinant protein production (de la Broise et al, 1991).…”

Potential of cell retention techniques for large‐scale high‐density perfusion culture of suspended mammalian cells

“…A single publication reports of a measured average residence time of 1.5 h (Searles et al, 1994). Settlers were reviewed earlier and compared with filters and centrifuges as retention devices for animal cell perfusion cultures (Tokashiki and Takamatsu, 1993). They were also used for other biological systems like yeast and plant cells (Maia and Nelson, 1993;Su et al, 1996).…”

“…The topic of mammalian cell retention techniques for perfusion cultures has been reviewed periodically (Scheirer, 1988;Tokashiki and Takamatsu, 1993;Woodside et al, 1998;Castilho and Medronho, 2002). The objective of this review is to focus on the development of large-scale bioreactor systems for suspended mammalian cells and to the applicable cell retention strategies.…”