Serum- and protein-free media formulations for the Chinese hamster ovary cell line DUKXB11

Schröder, Martin; Matischak, Kathrin; Friedl, Peter

doi:10.1016/j.jbiotec.2003.12.005

Cited by 48 publications

(45 citation statements)

References 63 publications

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“…This effect was cell line specific however, as CHO-K1 grew in insulin-free medium with zinc (marginally slower than with insulin) but the recombinant clone CHO-IFN could not. An alternative strategy to remove the requirement of insulin was used by Schröder et al (2004). By weaning CHO-DU-KXB11 cells from SFM with reducing insulin concentrations, a PFM was generated.…”

Section: Insulinsupporting

confidence: 81%

The role of recombinant proteins in the development of serum-free media

2006

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Early developments in serum-free media led to a variety of formulations in which components normally provided in serum and required for growth (insulin, transferrin, lipid supplements, trace elements) and poorly defined components (extracts, hydrolysates) were added to defined basal media. These additives were mostly animal-derived. Given recent concerns about TSEs (transmissible spongiform encephalopathies) and other adventitious agents, the drive in media formulations must be towards elimination of animal-origin materials while maintaining cell line productivity. The progress made towards removing animal-derived components and the use of recombinant proteins in serum-free media for mammalian cells is reviewed.

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Section: Insulinsupporting

confidence: 81%

The role of recombinant proteins in the development of serum-free media

2006

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“…As the shear protective effect of PF-68 has been reported to be proportional to its concentration (Ma et al, 2004;Zhang et al, 1992), it is important to know if this uptake phenomenon can significantly affect the Pluronic content in the culture medium. For CHO cells, a 0.2% (w/v) PF-68 concentration is commonly used (Keane et al, 2003;Schroder et al, 2004) and we showed that the presence of cells only slightly affects this concentration. At 0.2% (w/v) with 4 Â 10 6 cells/mL, only a 2.4% decrease of the initial concentration was observed, after 18 h. At the end of a classical bioreactor culture, cell densities can reach up to 10 Â 10 6 cells.…”

Section: Discussionmentioning

confidence: 99%

“…Each Pluronic fits for a specific application, from drug solubilization to bioprocess applications. For instance, Pluronic F-68 (PF-68, (EO) 75 -(PO) 30 -(EO) 75 ) is used as a shear protective agent which limits cell death in mechanically agitated gas-sparged bioreactors (Murhammer and Goochee, 1990) and is particularly useful in serum-free medium culture (Keane et al, 2003;Schroder et al, 2004). The protective effect of PF-68 has been demonstrated for suspension culture of mammalian cells (Jordan et al, 1994;Ramirez and Mutharasan, 1990;Zhang et al, 1992), insect cells (Goldblum et al, 1990;Murhammer and Goochee, 1990), and plant cells (Sowana et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

The fate of Pluronic F‐68 in chondrocytes and CHO cells

Gigout

Buschmann

Jolicœur

2008

Biotech & Bioengineering

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The surfactant Pluronic F-68 (PF-68) is widely used in large-scale mammalian cell culture to protect cells from shear stress that arises from agitation and gas sparging. Several studies suggested that PF-68 is incorporated into the cell plasma membrane and could enter the cells, but without providing any direct evidence. The current study has examined this question for two cell types, one of pharmaceutical interest (CHO cells) and the other of biomedical interest (chondrocytes or cartilage cells). A fluorescent derivative of PF-68 was synthesized to detect and localize internalized Pluronic with culture time. PF-68 uptake by the cells was quantified and characterized. We clearly demonstrate that PF-68 enters the cells, and possibly accumulates in the endocytic pathway. CHO cells showed an average uptake of 11.7 +/- 6.7 (SEM) microg PF-68/10(6) cells while the uptake of chondrocytes was 56.0 +/- 10.9 (SEM) microg PF-68/10(6) cells, independently of the initial PF-68 concentration (between 0.01 and 0.2%, w/v) and of cell concentration (from 1 x 10(6) to 4 x 10(6) cells/mL). These uptake values were identical for both static and agitated culture conditions. Finally, we found that CHO cells are able to eliminate intracellular fluorescent PF-68 but chondrocytes are not. These results show that the uptake of PF-68 by the cells can severely affect PF-68 concentration in the culture medium and thus shear protection effect.

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“…For decades, animal-derived serum has been an essential component of mammalian cell culture media (Hu and Aunins, 1997;Schroder et al, 2004). However, a number of issues associated with the use of serum make it preferable to propagate cells in serum-free media (SFM) (Schroder et al, 2004;Sinacore et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Egr1 and Gas6 facilitate the adaptation of HEK‐293 cells to serum‐free media by conferring enhanced viability and higher growth rates

Jaluria

Κωνσταντόπουλος

Betenbaugh

et al. 2007

Biotech & Bioengineering

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Animal-derived serum is an essential media supplement for mammalian cells in cell culture. For a number of reasons including cost, regulatory concerns, lot inconsistency, potential contamination with adventitious agents, and down-stream processing it is desirable to eliminate the use of serum. Existing protocols designed to adapt cells to serum-free media (SFM) are time-consuming and provide little insight into how the cells adapt. To better understand the physiological responses associated with serum withdrawal and to expedite the adaptation process, a Human Embryonic Kidney-293 (HEK-293) cell line was propagated in 10% fetal bovine serum (FBS) and was progressively adapted to SFM and analyzed at specific serum levels by oligonucleotide microarrays. Of the differentially expressed genes two, early growth response 1 (egr1) and growth arrest specific 6 (gas6), were selected for further analysis based on their level of differential expression, overall expression patterns, and proposed functionalities. HEK-293 cells, propagated in 10% FBS were transfected with egr1 or gas6 and then adapted to SFM. Results indicated that higher expression of either gene moderately enhanced the ability of both cell lines to adapt to SFM. Egr1 appeared to have a greater impact on adaptability than gas6. Results also indicated that specific protein production was unaltered when the expression of egr1 was increased. Flow cytometric analysis revealed increased expression of egr1 was associated with an increase in the percentage of cells in the G2/M phases. These results indicate that enhanced expression of egr1 or gas6 facilitate adaptation to SFM by improving growth and viability.

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Serum- and protein-free media formulations for the Chinese hamster ovary cell line DUKXB11

Cited by 48 publications

References 63 publications

The role of recombinant proteins in the development of serum-free media

The role of recombinant proteins in the development of serum-free media

The fate of Pluronic F‐68 in chondrocytes and CHO cells

Egr1 and Gas6 facilitate the adaptation of HEK‐293 cells to serum‐free media by conferring enhanced viability and higher growth rates

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