The effects of microcarrier culture on recombinant CHO cells under biphasic hypothermic culture conditions

Abstract

A Chinese hamster ovary (CHO) cell line, producing recombinant secreted human placental alkaline phosphatase (SEAP) was investigated under three different culture conditions (suspension cells, cells attached to Cytodex 3 and Cytopore 1 microcarriers) in a biphasic culture mode using a temperature shift to mild hypothermic conditions (33 degrees C) in a fed-batch bioreactor. The cell viability in both the suspension and the Cytodex 3 cultures was maintained for significantly longer periods under hypothermic conditions than in the single-temperature cultures, leading to higher integrated viable cell densities. For all culture conditions, the specific productivity of SEAP increased after the temperature reduction; the specific productivities of the microcarrier cultures increased approximately threefold while the specific productivity of the suspension culture increased nearly eightfold. The glucose and glutamine consumption rates and lactate and ammonia production rates were significantly lowered after the temperature reduction, as were the yields of lactate from glucose. However, the yield of ammonia from glutamine increased in response to the temperature shift.

Enhanced Production of Human Recombinant Proteins from CHO cells Grown to High Densities in Macroporous Microcarriers

“…The range of Q glc values measured (0.4-0.7 pg/ cell/day) compares with other reports for CHO cells in suspension as 0.54 pg/cell/day [42] or 0.99 pg/cell/day [43], and 1.21 pg/cell/day or 1.04 pg/cell/day for Cytopore 1 cultures [44]. Similarly, the measured Q lac values (0.22-0.58 pg/cell/day) are in range of comparable rates reported previously; 0.2 pg/cell/day [42], 0.84 pg/cell/day [43], 1.10 pg/cell/day in CHO suspension cultures and 1.02 pg/cell/day in a Cytopore 1 culture [44].…”

“…Although Nam et al [44] reported no apparent difference in metabolic rates between suspension and microcarrier cultures, we report significantly higher specific glucose consumption and lactate production rates measured in all the Cytopore cultures compared to suspension cultures. Furthermore, the Q glc values show a positive correlation with the cell densities within the microcarriers.…”

Improved Bioprocess with CHO-hTSH Cells on Higher Microcarrier Concentration Provides Higher Overall Biomass and Productivity for rhTSH

“…The CHO cells have been the cell substrate of choice for rhTSH preparation [4,7,9,10]. CHO cell lines are capable of growing as adherent cells on a large variety of supports as well as free in suspension [11][12][13][14]. These characteristics of CHO cell lines (adherent × suspension) determine quite specific and particular bioprocesses, which show advantages and disadvantages.…”

“…Since CHO cells have been the cell of choice to transfect and synthesize rhTSH [4,9,10], basically two main bioprocess technologies are available. They rely on a particular characteristic of these cells: growing attached to a surface or free in suspension [11][12][13][14]. These two technologies have been used and show advantages and disadvantages [15].…”

5.3 Nutrient Media for Cell Culture Technology

“…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. Applying a biphasic culture strategy which deploys temperature reduction in a fed-batch cultivation achieved significant improvements by extending the lifespan of the culture, increasing the integral of viable cell density (IVCD) also called the integral cumulative cell time by a factor of 1.6 and increasing the specific productivity of the recombinant protein by 2.3 times (see Figure 5.3.3) [91], resulting in a more efficient and productive process [84].…”