Kcm supplements

Manufactured by Thermo Fisher Scientific

Sourced in United States

KCM supplements are a line of laboratory equipment products manufactured by Thermo Fisher Scientific. These supplements are designed to support various laboratory applications, but a detailed and unbiased description of their core function cannot be provided without the risk of extrapolation or interpretation.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Tnf α, by Merck Group (2 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) Percoll, by Thermo Fisher Scientific (1 mentions) Interleukin 6 (il 6), by Merck Group (1 mentions) Il 10, by Merck Group (1 mentions) Dispase dnase, by Thermo Fisher Scientific (1 mentions) Anti cd90, by Agilent Technologies (1 mentions) Anti cd45, by Agilent Technologies (1 mentions) Hla g, by BD (1 mentions) Bq788, by Bio-Techne (1 mentions) Bq123, by Bio-Techne (1 mentions)

2 protocols using kcm supplements

Isolation and Culture of First Trimester Trophoblasts

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Human first trimester trophoblast were isolated as described elsewhere.²⁹ Briefly, placental villi were digested with Dispase/DNAse and Trypsin (Gibco, Invitrogen, Carlsbad, CA, USA). After Percoll (Gibco) gradient centrifugation, cells were incubated with magnetic beads conjugated with anti‐CD90 and anti‐CD45 (Dako, Glostrup, DK) to remove fibroblasts and common leukocyte‐antigen expressing cells. Purity of trophoblast isolations was determined by cytokeratin 7 (K7) (Dako, 1:750) and HLA‐G (BD‐Biosciences, Bedford, MA, USA, 1:500) immunostaining. Only preparations with a purity ≥ 95% were used.
Immediately after isolation, trophoblasts were seeded in gelatin pre‐coated plates and maintained for 48 hours in Keratinocyte medium (KCM, Gibco) with penicillin/streptomycin (Gibco), KCM supplements (Gibco), and 10% (v/v) fetal calf serum (FCS, Thermo Scientific, Scientific, Rockford, IL, USA) in a humidified incubator at 37°C, 5% CO₂. Subsequently, cells were cultured under low serum conditions (2% (v/v) FCS) for 24 hours and further incubated in the absence (control) or presence of IL‐6 (10 ng/mL, Sigma Aldrich, St. Louis, MO, USA), IL‐10 (50 ng/mL, Sigma Aldrich), or TNF‐α (10 ng/mL. Sigma Aldrich). Cytokine incubation was performed for 24 hours as previously demonstrated to be sufficient for regulation of MMPs.³⁰

Majali‐Martinez A., Hoch D., Tam‐Amersdorfer C., Pollheimer J., Glasner A., Ghaffari‐Tabrizi‐Wizsy N., Beristain A.G., Hiden U., Dieber‐Rotheneder M, & Desoye G. (2020). Matrix metalloproteinase 15 plays a pivotal role in human first trimester cytotrophoblast invasion and is not altered by maternal obesity. The FASEB Journal, 34(8), 10720-10730.

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Trophoblast Isolation and Treatment

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Isolated trophoblasts were seeded in gelatin pre-coated plates and maintained for 2 days in Keratinocyte medium (KCM, Gibco) with penicillin/streptomycin (Gibco), KCM supplements (Gibco) and 10% (v/v) fetal calf serum (FCS, Thermo Scientific) in a humidified incubator at 37°C, 5% CO₂ in air. Prior to treatments, the cells were cultured under low serum conditions (2% (v/v) FCS) for 24 h. Thereafter, the cells were incubated in the absence (control) or presence of 10 nM or 100 nM ET-1 (Sigma Aldrich, St. Louis, MO, USA) for 24 h. The ETR subtype involved in mediating ET-1 effects was determined by pre-incubating trophoblasts with selective ETR antagonists for 2 h prior to the addition of 100 nM ET-1: BQ-123 (Tocris, Bristol, UK, 1.4 and 11.2 nM) for ETRA and BQ-788 (Tocris, 1.2 and 9.6 nM) for ETRB. Because late first trimester (GW 11 + 12) placentas gave highest trophoblast yields, these isolations were used in this set of experiments. Trophoblasts were also treated with TNF-α (Sigma Aldrich, 25 ng/ml) either alone or in combination with 100 nM ET-1, or incubated in the absence or presence of 100 nM ET-1 under three different oxygen tensions (1%, 2.5% and 20% O₂) for 24 h.

Majali-Martinez A., Velicky P., Pollheimer J., Knöfler M., Yung H.W., Burton G.J., Tabrizi-Wizsy N.G., Lang U., Hiden U., Desoye G, & Dieber-Rotheneder M. (2016). Endothelin-1 down-regulates matrix metalloproteinase 14 and 15 expression in human first trimester trophoblasts via endothelin receptor type B. Human Reproduction (Oxford, England), 32(1), 46-54.

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