Best practices for media selection for mammalian cells

Price, Paul J.

doi:10.1007/s11626-017-0186-6

Cited by 47 publications

(39 citation statements)

References 19 publications

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“…Relevant in vitro models include the use of human primary fibroblasts (HFs) and endothelial cells (ECs) embedded in three-dimensional (3D) substrates (DeCicco-Skinner et al, 2014;Liu et al, 2017). A variety of endothelial culture media are reported (Price, 2017) to be enriched with varying concentrations of serum and synthetic growth factors, such as VEGF, fibroblast growth factor (FGF), insulin growth factor (IGF) and EGF. Although the methods mentioned previously provide conditions for the formation of vascular structures, the synthetic combination of growth factors (GFs) in a culture medium lacks the complexity and repertoire of proteins secreted naturally (secretome) into the extracellular space (Veronesi et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Platelet-rich fibrin secretome induces three dimensional angiogenic activation in vitro

Herrera-Vizcaíno¹,

Dohle²,

Al‐Maawi³

et al. 2019

eCM

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Different tissue engineering techniques are used to support rapid vascularisation. A novel technique is the use of platelet-rich fibrin (PRF), an autologous source of growth factors. This study was the first to investigate the influence of PRF matrices, isolated following different centrifugation protocols, on human dermal vascular endothelial cells (ECs) in mono-culture and co-culture with human primary fibroblasts (HFs) as an in vitro model for tissue regeneration. Focus was placed on vascular structure formation and growth factor release. HFs and ECs were cultivated with PRF prepared using a high (710 ×g) or low (44 ×g) relative centrifugation force (RCF) over 14 d. Immunofluorescence staining and immunohistochemistry were used to evaluate the microvascular formation. Cell culture supernatants were collected for evaluation of growth factor release. The results showed a PRF-mediated effect on the induction of angiogenesis in ECs. Microvessel-like structure formation was promoted when ECs were combined with low-RCF PRF as compared to high-RCF PRF or control group. The percentage of vascular lumen area was significantly higher in low-RCF PRF, especially at day 7, which coincided with statistically significantly higher growth factor [vascular endothelial factor (VEGF), transforming growth factor β1 (TGF-β1) and platelet derived growth factor (PDGF)] concentration measured in low-RCF PRF as compared to high-RCF PRF or control group. In conclusion, reducing the RCF according to the low-speed centrifugation concept (LSCC) resulted in increased growth factor release and angiogenic structure formation with EC mono-culture, suggesting that PRF may be a highly beneficial therapeutic tool for tissue engineering applications.

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Section: Introductionmentioning

confidence: 99%

Platelet-rich fibrin secretome induces three dimensional angiogenic activation in vitro

Herrera-Vizcaíno¹,

Dohle²,

Al‐Maawi³

et al. 2019

eCM

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“…FBS contains a wide array of mitogenic growth factors that are capable of promoting the survival and growth of cells in culture. ( 4 ) Indeed, FBS supplementation of hybridoma culture media is sufficient for generation of hybridoma cell lines. However, the use of MCM likely provides additional factors that facilitate growth and stability of more newly fused, polyploidal hybrid cells.…”

Section: Discussionmentioning

confidence: 99%

“…However, there remains a need for cost-effective, animal-derived culture supplements to support early hybridoma cell survival, cloning, and growth. ( 4 , 5 ) Indeed, newly formed hybrid cells often require the addition of a cell feeder layer (i.e., macrophages) or supplementation with a cell-conditioned medium for initial stabilization and growth. ( 6–8 ) The use of a cell feeder layer imposes several disadvantages that include interference with hybridoma cell growth and introduction of potential contamination.…”

Section: Introductionmentioning

confidence: 99%

“…Efforts have been made to identify defined medium conditions to support hybridoma growth and increase the efficiency of antibody production. ( 4 , 15 , 16 ) Proteins such as interlukin-1 (IL-1), tumor necrosis factor, ( 17 ) granulocyte–macrophage colony-stimulating factor, ( 18 ) and IL-6 ( 16 , 19 ) have been shown to support hybridoma growth, resulting in proprietary blends of defined hybridoma growth media. Although these commercial products can be effective, they can be cost prohibitive, and many laboratories rely on in-house production of macrophage-conditioned medium (MCM) to support hybridoma growth during early cell selection and cloning.…”

Section: Introductionmentioning

confidence: 99%

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A Bioassay for Optimization of Macrophage-Conditioned Medium as a Culture Supplement to Promote Hybridoma Cell Survival and Growth

Hnasko

Lin

Stanker

et al. 2018

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

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Macrophage-conditioned medium (MCM) is an important cell culture supplement used to support the survival and growth of newly fused hybridoma cells. The use of macrophage cells, as a part of hybridoma technology, has proven to be an effective and inexpensive source of growth factors that promote the early survival and growth of hybridoma cells. Despite the widespread use of MCM as a hybridoma culture supplement, there is limited guidance and standardization for MCM production to achieve optimal hybridoma support. As an undefined supplement, significant variations in production of MCM may negatively impact hybridoma cell survival and growth. The lack of an available method for standardization of MCM bioactivity has limited validation, optimization, and commercial production. Consequently, variations in batch production of MCM may result in low-quality MCM that limits hybridoma viability and negatively impacts monoclonal antibody production. In this report, we describe a novel bioassay based on the newly generated, MCM-dependent RMH359 hybridoma cell line that can be used to validate MCM bioactivity and standardize production. We demonstrate the utility of the RMH359 bioassay (1) for evaluating MCM hybridoma bioactivity, (2) to define optimal conditions for production of MCM, and (3) as a method for MCM validation and standardization. In conclusion, the RMH359 cell bioassay provides a specific and sensitive assessment of MCM bioactivity in support of hybridoma cell survival and growth.

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“…Murine-derived feeder cells are widely used to maintain the pluripotency of hiPSCs, and human-derived feeder cells are also used. However, these cells are unsuitable for stem cell maintenance (Price 2017) as the feeder cells are cultured in medium supplemented with fetal bovine serum or proprietary serum replacements. The use of culture medium containing undefined or unknown components has limited the understanding of development and cellular differentiation (Nims and Harbell 2017).…”

Section: Introductionmentioning

confidence: 99%

Induction of integration-free human-induced pluripotent stem cells under serum- and feeder-free conditions

Hamada

Akagi

Yamasaki

et al. 2019

In Vitro Cell.Dev.Biol.-Animal

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Human-induced pluripotent stem cells (hiPSCs) have shown great potential toward practical and scientific applications. We previously reported the generation of human dental pulp stem cells using non-integrating replication-defective Sendai virus (SeVdp) vector in feeder-free culture with serum-free medium hESF9. This study describes the generation of hiPSCs from peripheral blood mononuclear cells to increase the donor population, while reducing biopsy invasiveness. From 6-d-old primary culture of peripheral blood mononuclear cells (PBMCs) with IL-2, hiPSCs were established using SeVdp(KOSM)302L with recombinant Laminin-511 E8 fragments under serum-free condition. The established PBMC-derived hiPSCs showed pluripotency and differentiation ability both in vivo and in vitro. In addition, we evaluated microarray data from PBMC-and dental pulp-derived hiPSCs. These hiPSCs will be beneficial for characterizing the molecular mechanisms of cellular differentiation and may provide useful substrates for developing cellular therapeutics.

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Best practices for media selection for mammalian cells

Cited by 47 publications

References 19 publications

Platelet-rich fibrin secretome induces three dimensional angiogenic activation in vitro

Platelet-rich fibrin secretome induces three dimensional angiogenic activation in vitro

A Bioassay for Optimization of Macrophage-Conditioned Medium as a Culture Supplement to Promote Hybridoma Cell Survival and Growth

Induction of integration-free human-induced pluripotent stem cells under serum- and feeder-free conditions

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