Normal human dermal fibroblasts (nhdf)

Manufactured by Cambrex

Sourced in United Kingdom

NHDF is a laboratory equipment product manufactured by Cambrex. It serves as a core function for various research and testing applications.

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Lab products found in correlation

Dmem f12, by Corning (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Human umbilical vein endothelial cells (huvec), by Thermo Fisher Scientific (1 mentions) Human umbilical vein endothelial cells (huvec), by Cambrex (1 mentions) Human umbilical vein endothelial cells (huvec), by Lonza (1 mentions) Normal human dermal fibroblasts (nhdf), by Lonza (1 mentions) Normal human dermal fibroblasts (nhdf), by Thermo Fisher Scientific (1 mentions) Egm 2 singlequots, by Cambrex (1 mentions) Human mesenchymal stem cells, by Lonza (1 mentions) Mscgm, by Lonza (1 mentions) Hmscs, by Lonza (1 mentions) Dmem f12, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Mouse anti β actin, by Cell Signaling Technology (1 mentions) Mouse anti flag, by Merck Group (1 mentions) Rabbit anti vdac, by Merck Group (1 mentions)

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NHDFs (4 citations)

4 protocols using normal human dermal fibroblasts (nhdf)

Endothelial-Fibroblast Co-Culture Protocol

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All chemicals and cell culture reagents were purchased from Sigma-Aldrich Ltd. (Dorset, UK), unless otherwise stated.
Cell culture. Primary human umbilical vein endothelial cells (HUVEC) and primary normal human dermal fibroblasts (NHDF) were obtained from Clonetics (Lonza, Slough, UK). HUVEC were maintained in endothelial basal medium (EBM-2) supplemented with EGM-2 SingleQuots (i.e., EGM-2 medium) whilst NHDF were maintained in fibroblast basal medium (FBM) supplemented with FGM-2 SingleQuots; all from Clonetics; Lonza (i.e., FGM-2 medium).
The co-culture system was set up using HUVEC (between passages 1-8) and NHDF (between passages 1-12) following the protocol established by Bishop et al (15) and adapted by Barron et al (14) . HUVEC and NHDF were mixed and seeded in 24-well plates (Thermo Fisher Scientific Nunc, Loughborough, UK) in EGM-2 medium. Co-cultured cells were incubated for up to 14 days at 37˚C in a 5% Co 2 in air humidified incubator.

Barron G.A., Goua M., Wahle K.W.J, & Bermano G. (2017). Circulating levels of angiogenesis-related growth factors in breast cancer: A study to profile proteins responsible for tubule formation. Oncology reports, 38(3).

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Culturing Neonatal and Mesenchymal Stem Cells

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Neonatal human dermal fibroblast (nhDF, Clonetics) were maintained in a 50/50 mixture of Dulbecco’s Modification of Eagle’s Medium and Ham’s F12 cell culture medium (DMEM/F12, Cellgro) supplemented with 15% fetal bovine serum (FBS, Thermo-Fisher Scientific), 100 U/ml penicillin, and 100 μg/ml streptomycin. Cells were incubated at 37°C in 100% humidity and 5% CO2, passaged at ~90% confluency, and harvested for use at passage 9.
Human mesenchymal stem cells (Lonza) from a male donor aged 43 years old were maintained in mesenchymal stem cell growth media (MSCGM, Lonza). Cells were incubated at 37°C in 100% humidity and 5% CO2, passaged at ~80% confluency, and harvested for use at passage 6–7.

Weidenhamer N.K., Moore D.L., Lobo F.L., Klair N.T, & Tranquillo R.T. (2015). Influence of culture conditions and extracellular matrix alignment on human mesenchymal stem cells invasion into decellularized engineered tissues. Journal of Tissue Engineering and Regenerative Medicine, 9(5), 605-618.

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Neonatal Dermal Fibroblasts and hMSCs Maintenance

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Neonatal human dermal fibroblast (nhDF; Clonetics) were maintained in a 50:50 mixture of Dulbecco's modified Eagle's medium and Ham's F12 cell culture medium (DMEM/F12; Cellgro) supplemented with 15% fetal bovine serum (FBS; Thermo‐Fisher Scientific), 100 U/ml penicillin and 100 µg/ml streptomycin. Cells were incubated at 37 °C in 100% humidity and 5% CO₂, passaged at ~90% confluence and harvested for use at passage 9 (P9).
hMSCs (Lonza) from a male donor aged 43 years were maintained in mesenchymal stem cell growth medium (MSCGM; Lonza). The cells were incubated at 37 °C in 100% humidity and 5% CO₂, passaged at ~80% confluence and harvested for use at passage 6–7.

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Culturing Human Cell Lines and Antibody Detection

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The human osteosarcoma cell line U2OS and human embryonic kidney cell line HEK293 were grown in Dulbecco's modified Eagle's medium (DMEM, GIBCO) supplemented with 10% fetal bovine serum (FBS, Hyclone) at 37°C in humidified 5% CO₂ incubator. Human dermal fibroblasts derived from normal (NHDF, Clonetics) and XPD patient cells (GM00434) were grown in Dulbecco's Modified Eagle Medium/Ham's F-12 (DMEM/F-12; GIBCO) supplemented with 15% FBS, non-essential amino acids, essential amino acids, L-Glutamine, Vitamins and sodium pyruvate (GIBCO).
The following primary antibodies used for western blotting analysis were as follows: mouse anti-Flag (sigma), rabbit anti-XPD (Cell Signaling), goat anti-Lamin B for detecting nuclear protein (Santa Cruz); mouse anti-GAPDH for detecting cytoplasmic protein (Millipore); rabbit anti-VDAC for mitochondrial outer-membrane protein, mouse anti-SMAC for mitochondrial intermembrane space protein, rabbit anti-TUFM (Sigma) and mouse anti-β-actin (Cell Signaling).

Liu J., Fang H., Chi Z., Wu Z., Wei D., Mo D., Niu K., Balajee A.S., Hei T.K., Nie L, & Zhao Y. (2015). XPD localizes in mitochondria and protects the mitochondrial genome from oxidative DNA damage. Nucleic Acids Research, 43(11), 5476-5488.

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