Fetal clone 3

Manufactured by GE Healthcare

Sourced in Japan

The Fetal Clone III is a medical laboratory equipment designed for fetal DNA analysis. It features advanced genetic analysis capabilities to support prenatal testing and screening procedures.

Automatically generated - may contain errors

Lab products found in correlation

Trypsin edta, by Thermo Fisher Scientific (3 mentions) Insulin, by Merck Group (2 mentions) Penicillin streptomycin mixture, by Thermo Fisher Scientific (2 mentions) Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions) Mcf 7, by American Type Culture Collection (2 mentions) Coulter counter, by Beckman Coulter (2 mentions) Collagenase type 2, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Calcium and magnesium, by Thermo Fisher Scientific (1 mentions) Carnitine, by Merck Group (1 mentions) L serine, by Merck Group (1 mentions) Dmem ham s f12, by Thermo Fisher Scientific (1 mentions) Saos 2, by American Type Culture Collection (1 mentions) Sk lu 1, by American Type Culture Collection (1 mentions) Ht1080, by American Type Culture Collection (1 mentions) T0026, by Applied Biological Materials (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) 40 μm cell strainer, by Corning (1 mentions) Isoproterenol, by Merck Group (1 mentions) Epidermal growth factor (egf), by Merck Group (1 mentions)

8 protocols using fetal clone 3

Gingiva Equivalent Exposure to Oral Microbiomes

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Telomerase reverse transcriptase–immortalized human gingiva keratinocyte and
fibroblast cell lines were cultured and used for the construction of human
gingiva equivalents exactly as previously described, except that no antibiotics
were used in the culture media (Buskermolen et al. 2016 (link)). The commensal,
gingivitis, and cariogenic microbiomes, grown as described from a pool of 10
saliva donors, were diluted in Hank’s Balanced Salt Solution (HBSS) with calcium
and magnesium (Gibco) to 10⁷, 10⁸, and 10⁹CFUs/mL. Each concentration (10 µL) was dripped onto the surface of the gingiva
equivalents, for a final exposure of 10⁵, 10⁶, or
10⁷ CFUs/equivalent. Controls were exposed to 10 µL of HBSS.
Exposed gingiva equivalents were cultured by air exposure for 24 h at 37 °C,
7.5% CO₂, and 95% humidity on 1.5 mL of DMEM/Ham’s F12 (3/1; Gibco),
supplemented with 1% Fetal Clone III (GE), 0.1μM insulin (Sigma-Aldrich), 1μM
isoproterenol (Sigma-Aldrich), 10μM carnitine (Sigma-Aldrich), and 10mM L-serine
(Sigma-Aldrich). Each experiment was performed with an intraexperiment
duplicate. Three experiments were performed, each with a different batch of
gingiva equivalents, which were exposed to different batches of the cultured
biofilms, grown independently from the same pool of 10 saliva donors, as
described earlier.

Buskermolen J.K., Janus M.M., Roffel S., Krom B.P, & Gibbs S. (2017). Saliva-Derived Commensal and Pathogenic Biofilms in a Human Gingiva Model. Journal of Dental Research, 97(2), 201-208.

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Cell line culturing and transfection

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Cell lines (SKLU1, U2OS, SAOS-2, HELA, MCF7, HT1080) were obtained from American Type Culture Collection (ATCC). GM847 was obtained from the Coriell Institute cell repository. SUSM1 and HELA1.3 were obtained from Collins Lab (UC Berkeley, U.S.A.). Additional information of cell lines can be found in Supplementary Table S1. The cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 5% synthetic fetal bovine serum (FetalClone III; GE Life Sciences) and 1% penicillin-streptomycin mixture (Gibco). Incubation was done at 37°C in a humidified, 5% CO₂ atmosphere-controlled incubator (HERAcell). All cell culturing was performed using standard procedures, including aspiration and washing with phosphate-buffered saline (PBS, Gibco), trypsinizing using Trypsin-EDTA (Gibco). Coulter Counter (Beckman Coulter) was used for cell counting. Transfection protocols can be found in the Extended Methods (Supplementary File).

Yang S.Y., Chang E.Y., Lim J., Kwan H.H., Monchaud D., Yip S., Stirling P.C, & Wong J.M. (2021). G-quadruplexes mark alternative lengthening of telomeres. NAR Cancer, 3(3), zcab031.

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Isolation and Culture of Human Gingival Fibroblasts

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Healthy human gingiva tissue was used in an anonymous fashion in accordance with the “Human Tissue and Medical Research: Code of conduct for responsible use” as formulated by the Federation of Dutch Medical Scientific Societies (www.federa.org). Primary human gingiva fibroblasts were isolated as previously described (Buskermolen et al., 2016). In short, after enzymatic separation of the epithelium and lamina propria, the lamina propria was digested in collagenase type II (Gibco, Grand Island, NY) for 3 hr. After passing the tissue suspension through a 40 μm cell strainer (Corning, Oneonta, NY) and washing with PBS, the cells were cultured in fibroblast medium consisting of DMEM (Gibco), supplemented with 5% Fetal Clone III (GE, Logan, UT) and 1% penicillin‐streptomycin (Gibco). Fibroblasts were used between passage 2 and 4. The hTERT‐immortalized human gingiva fibroblasts (T0026, purchased from ABM, Richmond, BC, Canada) were also cultured in fibroblast medium and used until passage 30.

Buskermolen J.K., Roffel S, & Gibbs S. (2017). Stimulation of oral fibroblast chemokine receptors identifies CCR3 and CCR4 as potential wound healing targets. Journal of Cellular Physiology, 232(11), 2996-3005.

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Culturing MCF7 Human Breast Cancer Cells

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Human breast cancer cells MCF7 was obtained from American Type Culture Collection (ATCC). The cells were culture in Dulbecco's modified Eagle's medium (Gibco) supplemented with 5% synthetic fetal bovine serum (FetalClone III; GE Life Sciences) and 100 U penicillin–streptomycin mixture (Gibco). Cells were incubated at 37 °C in a humidified, 5% CO₂ atmosphere-controlled incubator (HERAcell). Standard cell-culturing procedures were employed including aspiration and washing with phosphate-buffered saline (PBS, Gibco). Cells were trypsinized using Trypsin-EDTA (0.25%) (Gibco). Cell counting was performed using a Coulter Counter (Beckman Coulter).

Yang S.Y., Lejault P., Chevrier S., Boidot R., Robertson A.G., Wong J.M, & Monchaud D. (2018). Transcriptome-wide identification of transient RNA G-quadruplexes in human cells. Nature Communications, 9, 4730.

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Isolation of Human Gingival Cells

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Healthy human gingiva tissue was used in an anonymous manner in accordance with the “Code for Proper Use of Human Tissues” as formulated by the Dutch Federation of Medical Scientific Organizations (www.fmwv.nl). Primary gingiva KC and fibroblasts were isolated as previously described for skin.^{26–28 (link)} In short, after overnight digestion in dispase II (Sigma-Aldrich) to separate the epithelial sheet from the lamina propria, gingiva KC (KC-Prim) were isolated from the epithelium with 0.05% trypsin/EDTA (Gibco) solution. KC were cultured in KC medium consisting of DMEM/Ham's F12 (3/1) (Gibco), supplemented with 5% Fetal Clone III (GE), 1% penicillin–streptomycin (Gibco), 1 μM hydrocortisone (Sigma-Aldrich), 0.1 μM insulin (Sigma-Aldrich), 1 μM isoproterenol (Sigma-Aldrich), and 1 ng/mL epidermal growth factor (EGF) (Sigma-Aldrich). KC-Prim were used until the third passage. Gingival fibroblasts (Fib-Prim) were isolated from the lamina propria by incubation in a collagenase type II (Gibco) solution and cultured in fibroblast medium consisting of DMEM, supplemented with 5% Fetal Clone III and 1% penicillin–streptomycin. The Fib-Prim were used between passage 3 and 4.

Buskermolen J.K., Reijnders C.M., Spiekstra S.W., Steinberg T., Kleverlaan C.J., Feilzer A.J., Bakker A.D, & Gibbs S. (2016). Development of a Full-Thickness Human Gingiva Equivalent Constructed from Immortalized Keratinocytes and Fibroblasts. Tissue Engineering. Part C, Methods, 22(8), 781-791.

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Chick Embryo Myoblast Isolation and Myostatin Treatment

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Breast muscles dissected from 14-day-old chick embryos were minced using surgical scissors and digested with HBSS (+) (Nacalai Tesque, Inc., Kyoto, Japan) containing 0.2% collagenase (Worthington Biochemical Corp., Lakewood, NJ, USA) for 20 min at 37°C. The cells were collected by centrifugation and resuspended in DMEM (Nacalai Tesque, Inc., Kyoto, Japan) supplemented with 15% serum (Fetal Clone III, GE Healthcare Bio-Sciences AB, Uppsala, Sweden), 1× non-essential amino acid solution (Nacalai Tesque, Inc., Kyoto, Japan), and 1× gentamicin/amphotericin solution (Life Technologies, Carlsbad, CA, USA). The cell suspension was passed through a cell strainer to remove tissue debris and then transferred to an uncoated flask to allow attachment of fibroblasts. After 1 h, the unattached cells were transferred to another uncoated flask and this procedure was repeated 2–3 times. The unattached cells were counted and plated onto collagen I-coated 12-well plates at 1×10⁵ cells/well. The cells were incubated in the medium described above at 37°C, with 5 % CO₂ in a humidified chamber until the formation of myotubes. Myotubes were treated with or without 20nM recombinant human/rat/mouse myostatin (PeptroTech Inc., Rocky Hill, NJ, USA) for 2 h, in the absence or presence of serum.

Saneyasu T., Honda K, & Kamisoyama H. (2019). Myostatin Increases Smad2 Phosphorylation and Atrogin-1 Expression in Chick Embryonic Myotubes. The Journal of Poultry Science, 56(3), 224-230.

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NSC34 and MEF Cell Culture Conditions

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All components necessary for cell culture were bought from Invitrogen unless stated otherwise. NSC34 cells (Cashman et al, 1992 (link)) and MEFs were cultured and maintained in DMEM containing 4.5 g/l glucose supplemented with 2 mM glutamine, penicillin (200 U/mL), and streptomycin (200 μg/ml), and 5% defined fetal bovine serum (GE Healthcare Hyclone) for NSC34 cells or 10% FetalClone III (GE Healthcare Hyclone). Pheonix-ECO HEK293 cells were cultured in DMEM supplemented with GlutaMAX, 10% fetal FCS, penicillin (200 U/ml), and streptomycin (200 μg/ml). For glucose and serum starvation, we omitted glucose or serum, respectively, and further supplemented the medium with pyruvate (1 mM) and nonessential amino acids. HBSS starvation was carried out with HBSS (with calcium and magnesium).

Tadepalle N., Robers L., Veronese M., Zentis P., Babatz F., Brodesser S., Gruszczyk A.V., Schauss A., Höning S, & Rugarli E.I. (2020). Microtubule-dependent and independent roles of spastin in lipid droplet dispersion and biogenesis. Life Science Alliance, 3(6), e202000715.

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SILAC Labeling of HeLa Cells

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HeLa cells were cultured at 37°C and 5% CO₂ in DMEM containing 4.5g/litre of glucose (# 11960044; Gibco), 2 mM glutamine (# 25030024; Gibco), 100 U/ml penicillin, 100 μg/ml streptomycin (# 15140122; Gibco), and 10% FetalClone III (#SH30109.03; GE Healthcare Hyclone). For SILAC labelling, cells were grown in arginine- and lysine-free DMEM with 4.5g/litre glucose (# 28001300; Silantes), containing dialysed FCS (#26400044; Gibco), penicillin–streptomycin–glutamine (# 10378016; Gibco), and different isotopes of arginine and lysine. We used medium isotopes (Arg6, Lys4) (# 201203902 and 211103913; Silantes) for the WT, light (Arg0, Lys0) (# 201003902 and 211003902; Silantes) for the DKO, and heavy (Arg10, Lys8) (# 201603902 and 211603902; Silantes) for the DKO^+E1/E2. Cells were regularly tested to be mycoplasma-free.

Veronese M., Kallabis S., Kaczmarek A.T., Das A., Robers L., Schumacher S., Lofrano A., Brodesser S., Müller S., Hofmann K., Krüger M, & Rugarli E.I. (2024). ERLIN1/2 scaffolds bridge TMUB1 and RNF170 and restrict cholesterol esterification to regulate the secretory pathway. Life Science Alliance, 7(8), e202402620.

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