Process Biotechnology

Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 10 No. 2, Issue of April 15, 2007
© 2007 by Pontificia Universidad Católica de Valparaíso -- Chile Received June 15, 2006 / Accepted December 20, 2006
DOI: 10.2225/vol10-issue2-fulltext-15
RESEARCH ARTICLE

Evaluating kinetic and physiological features of rCHO-K1 cells cultured on microcarriers for production of a recombinant metalloprotease/disintegrin

Kamilla Swiech*
Department of Chemical Engineering
Federal University of Sao Carlos
Via Washington Luiz, Km 235
Zip Code 13565-905
São Carlos-SP, Brazil
Tel: 55 16 3351 8264
Fax: 55 16 3351 8266
E-mail: kamillaswiech@hotmail.com

Gracinda M.C. da Silva
Department of Chemical Engineering
Federal University of Sao Carlos
Via Washington Luiz, Km 235
Zip Code 13565-905,
São Carlos-SP, Brazil
Tel: 55 16 3351 8264
Fax: 55 16 3351 8266
E-mail:castelo1@ig.com.br

Teresa C. Zangirolami
Department of Chemical Engineering
Federal University of Sao Carlos
Via Washington Luiz, Km 235
Zip Code 13565-905
São Carlos-SP, Brazil
Tel: 55 16 3351 8264
Fax: 55 16 3351 8266
E-mail: teresacz@power.ufscar.br 

Mônica R.C. Iemma
Department of Physiological Sciences
Federal University of Sao Carlos
Via Washington Luiz, Km 235
Zip Code 13565-905
São Carlos-SP, Brazil
Tel: 55 16 3351 8264
Fax: 55 16 3351 8266
E-mail: costaiemma@hotmail.com

Heloísa S. Selistre-de-Araújo
Department of Physiological Sciences
Federal University of Sao Carlos
Via Washington Luiz, Km 235
Zip Code 13565-905
São Carlos-SP, Brazil
Tel: 55 16 33518333
Fax: 55 16 33518401;
E-mail: hsaraujo@power.ufscar.br

Cláudio A.T. Suazo
Department of Chemical Engineering
Federal University of Sao Carlos
Via Washington Luiz, Km 235
Zip Code 13565-905
São Carlos-SP, Brazil
Tel: 55 16 3351 8264
Fax: 55 16 3351 8266
E-mail: claudio@power.ufscar.br

*Corresponding author

Financial support: Work supported by Fapesp (Proc. 01/02068-4) and CNPq scholarships (Proc.131241/2001-0).

Keywords: adhesion, animal cell, CHO-K1, growth, microcarrier, recombinant disintegrin.

Abbreviations:

ACLD: Agkistrodon contortrix lacticinctus
ADC: anchorage-dependent cells
SE: standard error

Abstract   Full Text

We present kinetic and physiological data regarding the culturing of rCHO-K1 cells on various microcarriers, to evaluate the potential of this culture strategy for mass production of these cells and expression of a recombinant disintegrin. Cultures were performed in 500 mL spinner flasks in DMEM culture medium with 10% v/v fetal calf serum, gently shaken at 37ºC, pH 7.4, in a 10% v/v CO2 atmosphere. The following values were obtained, respectively, for the adhesion time-constant Ka (h) and specific growth rate μmax (d-1) on each microcarrier: Cytodex 1 (0.91, 0.45), Cultispher S (0.28, 0.34), Immobasil FS (0.85, 0.52) and Pronectin F (5.12, 0.67). Metabolic characteristics showed some variation among the cultures with the four microcarriers, the most significant being the higher production of ammonia with microcarriers coated with adhesive molecules (Cultispher S and Pronectin F) relative to the uncoated carriers (Cytodex 1 and Immobasil FS). Experiments where the DMEM medium was gradually replaced by the serum-free medium (CHO-SFM-II) revealed important advantages over media containing serum, not only for assay purposes, but also for purification of the disintegrin. Altogether these results demonstrate that cultures on microcarriers, especially on Pronectin F, show good potential for larger scale cultures of rCHO-K1 cell.

Supported by UNESCO / MIRCEN network