Relationship between tissue plasminogen activator production and specific growth rate in Chinese Hamster Ovary cells cultured in mannose at low temperature

Abstract

Chinese hamster ovary (CHO) cells, producing human recombinant tissue plasminogen activator (tPA), were grown with mannose (5, 20 and 40 mM) instead of glucose at 31, 33 and 37 degrees C. The highest tPA concentration (1.5 mg l(-1) at 144 h of cultivation) and tPA specific production rate (47 ng 10(-6) cell h(-1)) were obtained at 31 degrees C and 40 mM mannose. Regardless of the temperature or mannose concentration used, an inverse relationship between the specific growth rate and tPA specific production rate was observed, suggesting that tPA production rate would be directly controlled by the growth rate.

Simultaneous environmental manipulations in semi-perfusion cultures of CHO cells producing rh-tPA

“…As shown in Table 1, regardless of the carbon source used, a decrease in the temperature of cultivation from 37 to 31ºC increased the culture longevity (defined as the time to reach 85% viability). Similar results have been previously reported for CHO cell cultures (Berrios et al 2009). In the CHO cultures with glucose, the qtPA increased when the cultivation temperature was decreased from 37 to 31ºC (Table 1), indicating that 31ºC was the most suitable condition for rh-tPA production.…”

“…Thus, an increase in qtPA was observed when the temperature was decreased from 37 to 34ºC, whereas the lowest qtPA value was observed at 31ºC. In contrast, Berrios et al (2009) have reported that the qtPA value can be enhanced in batch cultures of CHO cells conducted at 31ºC, using mannose as the carbon source. Furthermore, Berrios et al (2011) reported that replacing glucose by mannose in continuous culture, increased specific productivity of t-PA at high concentrations of the Cell growth and rh-tPA production in semi-perfusion cultures Figure 1 shows the profile of cell growth, cell viability and rh-tPA concentration in semi-perfusion cultures of CHO cells cultured at 37ºC with glucose as the main carbon source (SPC1).…”

“…However, the cultivation of these cells in bioreactors still has several areas to be optimized: cell growth, culture longevity, protein productivity, prevention of apoptosis induction and improve quality production (Mohan et al 2008). In an effort to increase the productivity of recombinant proteins, the effect of various environmental parameters, such as pH (Yoon et al 2005;Trummer et al 2006a), media formulation (Altamirano et al 2000;Berrios et al 2009) and temperature (Yoon et al 2003a;Yoon et al 2003b;Fox et al 2004;Yoon et al 2007), have been extensively investigated. CHO cell cultures that are utilized for the production and investigation of recombinant proteins are typically cultured in vitro at 37ºC.…”

Microarray and proteomics expression profiling identifies several candidates, including the valosin‐containing protein (VCP), involved in regulating high cellular growth rate in production CHO cell lines

“…A number of approaches have been taken to improve the productivity of mammalian cell lines through cell line engineering, including the direct manipulation of productivity (Peng and Fussenegger, 2009;Tigges and Fussenegger, 2006) as well as the suppression of apoptosis (Arden et al, 2007;Ifandi and Al-Rubeai, 2005). An important approach to achieving this aim is via manipulation of the cell-specific growth rate, especially given substantial evidence that the growth rate and specific production rate of CHO are directly linked (Berrios et al, 2009;Lipscomb et al, 2005;Yokota and Tanji, 2008). A highly desirable feature for any production cell line is an ability to grow quickly in the bioreactor, shortening time spent in the log phase of growth.…”

5.3 Nutrient Media for Cell Culture Technology

“…In this regard, 2 approaches have important implications on the culture performance. Changing the culture temperature to a mild hypothermia (33 to 30 °C) has a positive effect on the productivity of a wide range of recombinant proteins [89,90,3] in batch culture. A reduced temperature can positively affect the cellular metabolism, which then has an impact on Brought to you by | New York University Bobst Library Technical Services Authenticated Download Date | 7/6/15 11:59 AM the design of the feed medium in a fed-batch cultivation.…”