Characterization of growth and metabolism of Drosophila melanogaster cells transfected with the rabies-virus glycoprotein gene.
In the present study, the growth and key metabolic features of a gene-transfected Drosophila melanogaster (fruitfly) S2 (Schneider 2) cell population (S2AcRVGP cells), cultured in Sf900-II medium, have been evaluated to provide substantial support for the development of a bioprocess to produce RVGP (rabies-virus glycoprotein). Experimental cultures were grown both in a 100 ml Schott flask incubated in a shaker at 28 degrees C and 100 rev./min and in a 3 litre stirred-tank bioreactor at 28 degrees C, with increasing agitation. In small-scale culture, S2AcRVGP cells reached a maximum cell concentration of 1.13 x 10(7) cell/ml, presented a mu(max) (maximum specific growth rate) of 0.037 h(-1) and the growth was limited by oxygen deprivation. An early and remarkably long stationary phase was observed under hypoxia. Cell cultures grown in the bioreactor without oxygen limitation exhibited a maximum cell concentration of 2.2 x 10(7) cells/ml and mu(max) values as high as 0.048 h(-1). The main substrate consumed in order to reach such a high growth rate was the amino acid proline, which seems to play an important role as a source of metabolic energy in the culture of S2AcRVGP cells. Under conditions of hypoxia, the cells were able to survive for 15 h without apparent damage, recovering their previous metabolic activity.
Swiech
Kamilla
K
Departamento de Engenharia Química, Universidade Federal de São Carlos, Via Washington Luis, Km 235, CEP 13565-905, São Carlos-SP, Brazil.
da Silva
Clóvis S
CS
Arantes
Mabel K
MK
dos Santos
Alexandra S
AS
Astray
Renato M
RM
Pereira
Carlos A
CA
Suazo
Cláudio A T
CA
D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D003001:Cloning, Molecular; D004331:Drosophila melanogaster / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D011820:Rabies virus / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*
D006023:Glycoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins