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Cell type-specific modulation of cobalamin uptake by bovine serum

Journal Article


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Abstract


  • Tracking cellular &sup57;Co-labelled cobalamin (57Co-Cbl) uptake is a well-established method

    for studying Cbl homeostasis. Previous studies established that bovine serum is not generally

    permissive for cellular Cbl uptake when used as a supplement in cell culture medium,

    whereas supplementation with human serum promotes cellular Cbl uptake. The underlying

    reasons for these differences are not fully defined. In the current study we address this

    question. We extend earlier observations by showing that fetal calf serum inhibits cellular

    57Co-Cbl uptake by HT1080 cells (a fibrosarcoma-derived fibroblast cell line). Furthermore,

    we discovered that a simple heat-treatment protocol (95ÊC for 10 min) ameliorates this inhibitory

    activity for HT1080 cell 57Co-Cbl uptake. We provide evidence that the very high level

    of haptocorrin in bovine serum (as compared to human serum) is responsible for this inhibitory

    activity. We suggest that bovine haptocorrin competes with cell-derived transcobalamin

    for Cbl binding, and that cellular Cbl uptake may be minimised in the presence of large

    amounts of bovine haptocorrin that are present under routine in vitro cell culture conditions.

    In experiments conducted with AG01518 cells (a neonatal foreskin-derived fibroblast cell

    line), overall cellular 57Co-Cbl uptake was 86% lower than for HT1080 cells, cellular TC production

    was below levels detectable by western blotting, and heat treatment of fetal calf

    serum resulted in only a modest increase in cellular 57Co-Cbl uptake. We recommend a

    careful assessment of cell culture protocols should be conducted in order to determine the

    potential benefits that heat-treated bovine serum may provide for in vitro studies of mammalian

    cell lines.

Publication Date


  • 2016

Citation


  • Zhao, H., Ruberu, K., Li, H. & Garner, B. (2016). Cell type-specific modulation of cobalamin uptake by bovine serum. PLoS One, 11 (11), e0167044-1-e0167044-22.

Scopus Eid


  • 2-s2.0-84997190811

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=1965&context=ihmri

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/940

Start Page


  • e0167044-1

End Page


  • e0167044-22

Volume


  • 11

Issue


  • 11

Place Of Publication


  • United States

Abstract


  • Tracking cellular &sup57;Co-labelled cobalamin (57Co-Cbl) uptake is a well-established method

    for studying Cbl homeostasis. Previous studies established that bovine serum is not generally

    permissive for cellular Cbl uptake when used as a supplement in cell culture medium,

    whereas supplementation with human serum promotes cellular Cbl uptake. The underlying

    reasons for these differences are not fully defined. In the current study we address this

    question. We extend earlier observations by showing that fetal calf serum inhibits cellular

    57Co-Cbl uptake by HT1080 cells (a fibrosarcoma-derived fibroblast cell line). Furthermore,

    we discovered that a simple heat-treatment protocol (95ÊC for 10 min) ameliorates this inhibitory

    activity for HT1080 cell 57Co-Cbl uptake. We provide evidence that the very high level

    of haptocorrin in bovine serum (as compared to human serum) is responsible for this inhibitory

    activity. We suggest that bovine haptocorrin competes with cell-derived transcobalamin

    for Cbl binding, and that cellular Cbl uptake may be minimised in the presence of large

    amounts of bovine haptocorrin that are present under routine in vitro cell culture conditions.

    In experiments conducted with AG01518 cells (a neonatal foreskin-derived fibroblast cell

    line), overall cellular 57Co-Cbl uptake was 86% lower than for HT1080 cells, cellular TC production

    was below levels detectable by western blotting, and heat treatment of fetal calf

    serum resulted in only a modest increase in cellular 57Co-Cbl uptake. We recommend a

    careful assessment of cell culture protocols should be conducted in order to determine the

    potential benefits that heat-treated bovine serum may provide for in vitro studies of mammalian

    cell lines.

Publication Date


  • 2016

Citation


  • Zhao, H., Ruberu, K., Li, H. & Garner, B. (2016). Cell type-specific modulation of cobalamin uptake by bovine serum. PLoS One, 11 (11), e0167044-1-e0167044-22.

Scopus Eid


  • 2-s2.0-84997190811

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=1965&context=ihmri

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/940

Start Page


  • e0167044-1

End Page


  • e0167044-22

Volume


  • 11

Issue


  • 11

Place Of Publication


  • United States