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 |
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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
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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.
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