Properties of two insect cell lines useful for the baculovirus expression system in serum-free culture

Abstract

The properties of Sf9 and Tn5 insect cells were analyzed comparatively under serum‐free culture conditions. Sf9 cells in SF900II medium apparently utilized sucrose as a primary nutrient both before and after virus infection, yielding small amounts of lactate and ammonia. Tn5 cells in Excell 401 medium consumed all the nutrients examined, including sucrose. The productivity of a recombinant glycoprotein, OSF‐2, by Tn5 cells, was moderate in both monolayer and spinner cultures, but the ability to secrete it was compromised in the former case. Relative to the Tn5 cultures, Sf9 produced 30‐fold more OSF‐2 in either culture mode. © 1996 John Wiley & Sons, Inc.

Metabolomic study of baculovirus infected insect cells to identify metabolite based strategies for improving virus yields

“…Unlike Sf9 cells, HzAM1cells show a preference for asparagine over glutamine (Lua and Reid, 2003), and this is similar for the High Five cells. It has been reported for Sf9 cells that the production levels of lactate and ammonia are minimal even at high substrate concentrations (Bedard et al, 1993;Benslimane et al, 2005;Drews et al, 2000;Ferrance et al, 1993;Ikonomou et al, 2003;Rhiel et al, 1997;Sugiura and Amann, 1996;Wang et al, 1993). In contrast, HzAM1 cells showed a significant level of ammonia production in Sf900™II medium, which is an asparagine and glutamine rich medium (Lua and Reid, 2003).…”

“…Generally, there are some common aspects of insect cell metabolism including the utilization of sugars, peptides, amino acids, organic acids, vitamins, lipids and organic salts (Agathos 1991), and the production of alanine, lactate and ammonium ions as byproducts (Rhiel et al, 1997;Sugiura and Amann, 1996). Glucose is usually the preferred carbon and energy source of insect cells (Bedard et al, 1993;Rhiel et al, 1997).…”

“…The main by-product of insect cell metabolism is alanine (Bedard et al, 1993;Drews et al, 1995;Ohman et al, 1995;Reuveny et al, 1993), but no inhibitory effect related to alanine has been reported (Doverskog et al, 1997). Lactate and ammonia production levels are minimal even at high substrate concentrations and not inhibitory unless other culture conditions become adversely inhibitory (Bedard et al, 1993;Benslimane et al, 2005;Drews et al, 2000;Ferrance et al, 1993;Ikonomou et al, 2003;Rhiel et al, 1997;Sugiura and Amann, 1996;Wang et al, 1993). These are only some 12 common aspects of insect cell metabolism, the specific consumption level and preference reported for Sf9 cells that the production levels of lactate and ammonia are minimal even at high substrate concentrations (Bedard et al, 1993;Benslimane et al, 2005;Drews et al, 2000;Ferrance et al, 1993;Ikonomou et al, 2003;Rhiel et al, 1997;Sugiura and Amann, 1996;Wang et al, 1993).…”

“…In particular, the metabolism of Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tn5B1-4 or High Five) cell lines have been studied thoroughly (Bedard et al, 1993;Benslimane et al, 2005;Drews et al, 1995;Drews et al, 2000;Ferrance et al, 1993;Ohman et al, 1995;Reuveny et al, 1993;Rhiel et al, 1997;Sugiura and Amann, 1996), as these two cell lines have been extensively used for recombinant protein production. In addition to Sf9 and High Five cells, the metabolism of other cell lines including Helicoverpa zea (Lua and Reid, 2003) and Anticarsia gemmatalis (de Rezende et al, 2009;Gioria et al, 2006) have also been investigated to a significant degree.…”

“…In particular, the metabolism of Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tn5B1-4 or High Five) cell lines have been studied thoroughly (Bedard et al, 1993;Benslimane et al, 2005;Drews et al, 1995;Drews et al, 2000;Ferrance et al, 1993;Ohman et al, 1995;Reuveny et al, 1993;Rhiel et al, 1997;Sugiura and Amann, 1996), as these two cell lines have been extensively used for recombinant protein production. In addition to Sf9 and High Five cells, the metabolism of other cell lines including Helicoverpa zea (Lua and Reid, 2003) and Anticarsia gemmatalis (de Rezende et al, 2009;Gioria et al, 2006) have also been investigated to a significant degree.Generally, there are some common aspects of insect cell metabolism including the utilization of sugars, peptides, amino acids, organic acids, vitamins, lipids and organic salts (Agathos 1991), and the production of alanine, lactate and ammonium ions as byproducts (Rhiel et al, 1997;Sugiura and Amann, 1996). Glucose is usually the preferred carbon and energy source of insect cells (Bedard et al, 1993;Rhiel et al, 1997).…”