Autor: |
Núñez EG; Laboratório de Células Animais, Departamento de Engenharia Química, Escola Politécnica, Universidade de São Paulo, Av. Prof. Luciano Gualberto, Trav. 3, 380, Butantã, P.O. Box 61548, São Paulo, SP, 05508-900, Brazil, eutimiocu@yahoo.com., Leme J, de Almeida Parizotto L, Chagas WA, de Rezende AG, da Costa BL, Monteiro DC, Boldorini VL, Jorge SA, Astray RM, Pereira CA, Caricati CP, Tonso A |
Jazyk: |
angličtina |
Zdroj: |
Cytotechnology [Cytotechnology] 2014 Aug; Vol. 66 (4), pp. 605-17. Date of Electronic Publication: 2013 Jul 12. |
DOI: |
10.1007/s10616-013-9612-0 |
Abstrakt: |
This work focused on determining the effect of dissolved oxygen concentration (DO) on growth and metabolism of BHK-21 cell line (host cell for recombinant proteins manufacturing and viral vaccines) cultured in two stirred tank bioreactors with different aeration-homogenization systems, as well as pH control mode. BHK-21 cell line adapted to single-cell suspension was cultured in Celligen without aeration cage (rotating gas-sparger) and Bioflo 110, at 10, 30 and 50 % air saturation (impeller for gas dispersion from sparger-ring). The pH was controlled at 7.2 as far as it was possible with gas mixtures. In other runs, at 30 and 50 % (DO) in Bioflo 110, the cells grew at pH controlled with CO2 and NaHCO3 solution. Glucose, lactate, glutamine, and ammonium were quantified by enzymatic methods. Cell concentration, size and specific oxygen consumption were also determined. When NaHCO3 solution was not used, the optimal DOs were 10 and 50 % air saturation for Celligen and Bioflo 110, respectively. In this condition maximum cell concentrations were higher than 4 × 10(6) cell/mL. An increase in maximum cell concentration of 36 % was observed in batch carried out at 30 % air saturation in a classical stirred tank bioreactor (Bioflo 110) with base solution addition. The optimal parameters defined in this work allow for bioprocess developing of viral vaccines, transient protein expression and viral vector for gene therapy based on BHK-21 cell line in two stirred tank bioreactors with different agitation-aeration systems. |
Databáze: |
MEDLINE |
Externí odkaz: |
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