Performance evaluation of CHO-K1 cell in culture medium supplemented with hemolymph

Raffoul, Tássia; Swiech, Kamilla; Arantes, Mabel Karina; Sousa, Álvaro Paiva Braga de; Mendonça, Ronaldo Zucatelli; Pereira, Carlos Augusto; Suazo, Cláudio Alberto Torres

doi:10.1590/s1516-89132005000400011

Cited by 16 publications

(20 citation statements)

References 8 publications

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“…Stirred culture systems, such as the rotary RCCS™ bioreactor used by Chen et al (8) and the spinner flasks employed by Fok and Zandstra (9), represent a significant improvement in culture techniques compared to static systems, as they offer several advantages in the scale-up process, includ-A.M. Fernandes et al www.bjournal.com.br ing: 1) a controlled environment leading to a homogeneous culture, 2) simplicity of handling, and 3) less susceptibility to contamination (10). Indeed, bioreactors have been used in a variety of bioprocesses, such as cell expansion (11)(12)(13)(14) and protein and virus production (15)(16)(17).…”

Section: Introductionmentioning

confidence: 99%

Successful scale-up of human embryonic stem cell production in a stirred microcarrier culture system

Fernandes

Marinho

Sartore

et al. 2009

Braz J Med Biol Res

106

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Future clinical applications of human embryonic stem (hES) cells will require high-yield culture protocols. Currently, hES cells are mainly cultured in static tissue plates, which offer a limited surface and require repeated sub-culturing. Here we describe a stirred system with commercial dextran-based microcarriers coated with denatured collagen to scale-up hES cell production. Maintenance of pluripotency in the microcarrier-based stirred system was shown by immunocytochemical and flow cytometry analyses for pluripotency-associated markers. The formation of cavitated embryoid bodies expressing markers of endoderm, ectoderm and mesoderm was further evidence of maintenance of differentiation capability. Cell yield per volume of medium spent was more than 2-fold higher than in static plates, resulting in a significant decrease in cultivation costs. A total of 10 8 karyotypically stable hES cells were obtained from a unitary small vessel that needed virtually no manipulation during cell proliferation, decreasing risks of contamination. Spinner flasks are available up to working volumes in the range of several liters. If desired, samples from the homogenous suspension can be withdrawn to allow process validation needed in the last expansion steps prior to transplantation. Especially when thinking about clinical trials involving from dozens to hundreds of patients, the use of a small number of larger spinners instead of hundreds of plates or flasks will be beneficial. To our knowledge, this is the first description of successful scale-up of feeder-and Matrigel™-free production of undifferentiated hES cells under continuous agitation, which makes this system a promising alternative for both therapy and research needs.

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Section: Introductionmentioning

confidence: 99%

Successful scale-up of human embryonic stem cell production in a stirred microcarrier culture system

Fernandes

Marinho

Sartore

et al. 2009

Braz J Med Biol Res

106

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“…However, baculovirus infection induces oxidative stress and cell death, which limits recombinant protein production (Vieira et al 2006). Recently some studies have demonstrated the presence of pharmacologically active substances in hemolymph of Lonomia obliqua, which can enhance Sf-9 insect cell growth and culture longevity (Maranga et al 2003;Raffoul et al 2005;Souza et al 2005;Mendonça et al 2008;Greco et al 2009). In the present work, hemolymph (whole extract or a semi-purified fraction) was applied to improve BEVS/IC system by increasing rotavirus recombinant protein.…”

Section: Introductionmentioning

confidence: 99%

Improvement of recombinant protein production by an anti-apoptotic protein from hemolymph of Lonomia obliqua

et al. 2010

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Apoptosis is a major problem in animal cell culture during production of biopharmaceuticals, such as recombinant proteins or viral particles. In the present work baculovirus-insect cell expression system (BEVS/IC) is used as model to produce rotavirus like-particles, composed by three layers of three different viral proteins (VP2, VP6 and VP7). In this model baculovirus infection also induces host cell death. Herein a new strategy to enhance cell life span and to increase recombinant rotavirus protein production of BEVS/IC system was developed. This strategy relies on hemolymph from Lonomia oblique (total extracts or a semi-purified fraction) medium supplementation. The total extract and a purified fraction from hemolymph of Lonomia obliqua were able to protect Sf-9 cell culture against apoptosis triggered by oxidative stress (using the pro-oxidant agents tert butylhydroperoxide and hydrogen peroxide) and by baculovirus infection. Furthermore, hemolymph enhance final recombinant protein production, as it was observed by the increased amounts of VP6 and VP7, which were measured by the semi-quantitative western blot method. In conclusion, hemolymph medium supplementation can be a promising strategy to improve cell viability and productivity of recombinant protein in BEVS/IC system.

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“…These procedures aim to allow high cellular densities at low cost and with reduced operation complexity. Several studies have reported the presence of substances with pharmacological activity in the insect hemolymph (Shiotsuki et al 2000;Yamamoto et al 1999;Guerrero et al 1999;Jiang et al 1999;Rosenfeld and Vanderberg 1998;Hamdaoui et al 1998;Maranga et al 2003;Huberman et al 1979;Lin et al 1998;Kurata et al 1994;Jones et al 1993;Zhu et al 2000;Lowenberger et al 1999;Lamberty et al 1999;Gross et al 1998;Johns et al 1998;Lanz-Mendoza et al 1996;Peters et al 1993;Souza et al 2005;Raffoul et al 2005; Kanaya and Kobayashi 2000;Mendonça et al 2008). Kanaya and Kobayashi (2000) isolated and characterized a protein from silkworm hemolymph able to increase the activity of a recombinant protein (luciferase) by approximately 6,000 times.…”

Section: Introductionmentioning

confidence: 99%

Study of kinetic parameters for the production of recombinant rabies virus glycoprotein

et al. 2009

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Gene expression in insect cells is an advantageous system for recombinant protein production, mainly because of its capacity to produce complex proteins with correct post-translational modifications. Recently, we identified and purified a protein from Lonomia obliqua hemolymph able to increase the production of rabies virus glycoprotein, expressed in Drosophila melanogaster cells, by about 60%. In this work, the kinetic parameters for cell growth and recombinant rabies virus glycoprotein production were determined in cultures of transfected Drosophila melanogaster Schneider 2 (S2) cells expressing recombinant rabies virus glycoprotein (rRVGP), enriched and non-enriched with the hemolymph of Lonomia obliqua (Hb). The highest concentration of rRVGP was achieved at the beginning of the culture enriched with Hb, indicating that the cells produce greater amounts of rRVGP per cell (specific rRVGP concentration) at the early exponential growth phase. After day 8, a decrease in the concentration of rRVGP (ng/mL) was observed, probably due to protein decomposition. The average specific rRVGP production rate (mu(rRVGP)) was 30 ng rRVGP/10(7)cell.day, higher than that observed in the non-enriched culture.

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Performance evaluation of CHO-K1 cell in culture medium supplemented with hemolymph

Abstract: ABSTRACT

Cited by 16 publications

References 8 publications

Successful scale-up of human embryonic stem cell production in a stirred microcarrier culture system

Successful scale-up of human embryonic stem cell production in a stirred microcarrier culture system

Improvement of recombinant protein production by an anti-apoptotic protein from hemolymph of Lonomia obliqua

Study of kinetic parameters for the production of recombinant rabies virus glycoprotein

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