Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 Mar;59(2):135-41.
doi: 10.1007/s10616-009-9199-7. Epub 2009 May 30.

Effects of heat treatment and concentration of fish serum on cell growth in adhesion culture of Chinese hamster ovary cells

Masashi Fujiwara  1 , Ryohei Tsukada, Itaru Shioya, Mutsumi Takagi
Affiliations

Effects of heat treatment and concentration of fish serum on cell growth in adhesion culture of Chinese hamster ovary cells

Masashi Fujiwara et al. Cytotechnology. 2009 Mar.

Abstract

The effects of heat treatment and concentration of fish serum (FS) on cell growth and human granulocyte-macrophage colony-stimulating factor (hGM-CSF) production in an adhesion culture of recombinant Chinese hamster ovary (CHO) cells, DR1000L4N, were investigated. The addition of heat treated FS instead of non-heat-treated FS improved cell growth in terms of cell density, which reached 60% that in 10% fetal calf serum (FCS)-containing medium (FCS medium). A decrease in FS concentration from 10 to 1.25% markedly increased cell density, which was 79% that in 10% FCS medium. The combination of heat treatment at 56 degrees C and the addition of FS at a low concentration (1.25%) showed an additive effect on cell growth and resulted in the same cell density as that in 10% FCS medium, whereas the hGM-CSF concentration in the culture using FS-containing medium (FS medium) was approximately 50% that in 10% FCS medium. The total lipid concentration in FS was more than three fold that in FCS. The effect of decreasing FS concentration on cell growth may be due to the low lipid concentration in FS medium, because addition of the lipids extracted from FS to 10% FCS and 1.25% FS media markedly decreased cell density. Consequently, the addition of heat-treated FS at low concentrations to medium may be useful for the growth of CHO cells without FCS.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Effect of heat treatment of fish serum on adhesive growth of CHO cells. CHO DR1000L4N cells were cultivated in adhesion for 120 h in media containing 10% FCS, heat-treated 10% FS (Seriola quinqueradiata) (46, 51, 56, or 61 °C), non-heat-treated 10% FS (NT), or no serum (0%) after initial adhesion for 24 h in 10% FCS medium. Mean ± SD (n = 3)
Fig. 2
Fig. 2
Effect of concentration of fish serum on adhesive growth of CHO cells. CHO DR1000L4N cells were cultivated in adhesion for 120 h in media containing 10% FCS, FS (Seriola quinqueradiata) (1.25, 2.5, 5, 10, or 20%) or no serum (0%) after initial adhesion for 24 h in 10% FCS medium. Mean ± SD (n = 3)
Fig. 3
Fig. 3
Effects of heat treatment and concentration of FS and FCS on growth of CHO cells. CHO DR1000L4N cells were cultivated in adhesion for 120 h in medium containing FS (Seriola quinqueradiata) (circles) or FCS (squares) with (closed symbols) or without (open symbols) heat treatment (56 °C) at various concentrations after initial adhesion for 24 h in 10% FCS medium. Mean ± SD (n = 3)
Fig. 4
Fig. 4
Effects of heat treatment and concentration of fish serum from red seabream (Pagrus major) on growth of CHO cells. CHO DR1000L4N cells were cultivated for 120 h in medium containing FS (Pagrus major) with (closed bars) or without (open bars) heat treatment (56 °C) at various concentrations after initial adhesion for 24 h in 10% FCS medium. Mean ± SD (n = 3)
Fig. 5
Fig. 5
Influences of lipids extracted from fish serum on growth of CHO cells. CHO DR1000L4N cells were cultivated in adhesion for 120 h in media containing 10% FCS supplemented with lipids extracted from FS (Seriola quinqueradiata) (open circles), heat-treated 1.25% FS (Seriola quinqueradiata) supplemented with lipids (closed squares), and no serum supplemented with lipids (open triangles), after initial adhesion for 24 h in 10% FCS medium. For comparison, the same cultivations were performed in media with various concentrations of lipids originally contained in heat-treated FS (Seriola quinqueradiata) (open squares). Data represent the means of triplicate cultures
Fig. 6
Fig. 6
Time course of cell density and hGM-CSF concentration in adhesion culture of CHO cells using fish serum. a Cell density, b hGM-CSF concentration. CHO DR1000L4N cells were cultivated in adhesion for 168 h in media containing 10% FCS (open circles), 1.25% fish serum (Seriola quinqueradiata) with (closed squares) or without (open squares) heat treatment, and no serum (0%) (open triangles) after initial adhesion for 24 h in 10% FCS medium. Data represent the means of triplicate cultures
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. {'text': '', 'ref_index': 1, 'ids': [{'type': 'DOI', 'value': '10.1007/s00253-007-0897-9', 'is_inner': False, 'url': 'https://doi.org/10.1007/s00253-007-0897-9'}, {'type': 'PubMed', 'value': '17334756', 'is_inner': True, 'url': 'https://pubmed.ncbi.nlm.nih.gov/17334756/'}]}
    2. Fujiwara M, Tsukada R, Tsujinaga Y, Takagi M (2007) Fetal calf serum-free culture of Chinese hamster ovary cells employing fish serum. Appl Microbiol Biotechnol 75:983–987. doi:10.1007/s00253-007-0897-9 - PubMed
    1. {'text': '', 'ref_index': 1, 'ids': [{'type': 'DOI', 'value': '10.1677/joe.0.120R019', 'is_inner': False, 'url': 'https://doi.org/10.1677/joe.0.120r019'}, {'type': 'PubMed', 'value': '2926294', 'is_inner': True, 'url': 'https://pubmed.ncbi.nlm.nih.gov/2926294/'}]}
    2. Funkenstein B, Silgergeld A, Cavari B, Laron Z (1989) Growth hormone increases plasma levels of insulin-like growth factor (IGF-I) in a teleost, the gilthead seabream (Sparus aurata). J Endocrinol 120:R19–R21. doi:10.1677/joe.0.120R019 - PubMed
    1. {'text': '', 'ref_index': 1, 'ids': [{'type': 'DOI', 'value': '10.1074/jbc.272.11.7285', 'is_inner': False, 'url': 'https://doi.org/10.1074/jbc.272.11.7285'}, {'type': 'PubMed', 'value': '9054425', 'is_inner': True, 'url': 'https://pubmed.ncbi.nlm.nih.gov/9054425/'}]}
    2. Hata J-I, Takeo J, Segawa C, Yamashita S (1997) A cDNA encoding fish fibroblast growth factor-2, which lacks alternative translation initiation. J Biol Chem 272:7285–7289. doi:10.1074/jbc.272.11.7285 - PubMed
    1. {'text': '', 'ref_index': 1, 'ids': [{'type': 'DOI', 'value': '10.1006/gcen.2001.7677', 'is_inner': False, 'url': 'https://doi.org/10.1006/gcen.2001.7677'}, {'type': 'PubMed', 'value': '11589631', 'is_inner': True, 'url': 'https://pubmed.ncbi.nlm.nih.gov/11589631/'}]}
    2. Larsen DA, Beckman BR, Dickhoff WW (2001) The effect of low temperature and fasting during the winter on metabolic stores and endocrine physiology (insulin, insulin-like growth factor-I, and thyroxine) of coho salmon, Oncorhynchus kisutch. Gen Comp Endocrinol 123:308–323. doi:10.1006/gcen.2001.7677 - PubMed
    1. {'text': '', 'ref_index': 1, 'ids': [{'type': 'DOI', 'value': '10.1016/j.aquaculture.2005.05.030', 'is_inner': False, 'url': 'https://doi.org/10.1016/j.aquaculture.2005.05.030'}]}
    2. Nordgarden U, Hansen T, Hemre G-I, Sundby A, Bjornsson BT (2005) Endocrine growth regulation of adult Atlantic salmon in seawater: the effect of light regime on plasma growth hormone, insulin-like growth factor-I, and insulin levels. Aquaculture 250:862–871. doi:10.1016/j.aquaculture.2005.05.030