MSCs were isolated from the peripheral blood (PB) of six sheep that were not part of the wound model experimental design. From each animal, 100 ml of blood was taken from the jugular vein using a vacutainer containing the anticoagulant Li-heparin. The mononuclear cells were isolated using the protocol of Martinello et al. [13 (link)]. Briefly, the blood was diluted 1:1 with PBS (phosphate-buffered saline) and placed on Ficoll-paque solution (Amersham Biosciences) to obtain mononuclear cells in interphase after centrifugation. Cultures were maintained at 37 °C with 5% CO2 in growth medium (DMEM 5671, Sigma-Aldrich) with 10% FCS (fetal bovine serum, Euroclone). On the day of application, PB-MSCs were trypsinized with 0.25% trypsin-EDTA (Euroclone, Italy) and resuspended in hyaluronic acid (Hyalgan®, Fidia).
Dmem 5671
Manufactured by Merck Group
DMEM 5671 is a cell culture medium formulated by Merck Group. It is designed to support the growth and maintenance of a variety of cell types in vitro. The medium provides essential nutrients, vitamins, and other components necessary for cell proliferation and survival.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin edta, by Euroclone (1 mentions)
Fetal calf serum (fcs), by Euroclone (1 mentions)
Ficoll paque solution, by Cytiva (1 mentions)
Dmem no glucose, by Thermo Fisher Scientific (1 mentions)
P2256, by Merck Group (1 mentions)
Hepes, by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
L glutamine, by Merck Group (1 mentions)
Sodium pyruvate, by Merck Group (1 mentions)
Begm media, by Lonza (1 mentions)
4 protocols using dmem 5671
1
Isolation and Preparation of Ovine Mesenchymal Stem Cells
2
Cell Culture Protocol for SH-SY5Y, HT22, and MEFs
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Cells SH-SY5Y, HT22 and MEFs cells were grown in DMEM (DMEM 5671, Sigma), supplemented with 10% FCS, L-glutamine (2 mM), penicillin (100 mg/ml) and streptomycin (100 mg/ml). Human fibroblasts from Coriell Institute for medical research (FAD-PS2-N141I (AG09908) and control fibroblasts (AG08525, AG08269, AG09173)) were grown in the same medium, but containing 15% FCS. Cells were grown in a humidified atmosphere containing 5% CO 2 and seeded onto glass coverslips (13, 18 or 24 mm diameter). Where indicated, galactose-containing medium was used: DMEM no glucose (GIBCO, 11966025), supplemented with 10 mM galactose, 5% FCS, L-glutamine (2 mM), penicillin (100 U/ml) and streptomycin (100 mg/ml).
3
Culturing Murine Fibroblasts in DMEM
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Cells from a murine fibroblast cell line (L929, 85011425; Sigma-Aldrich) were
cultured and expanded in Dulbecco's modified Eagle's medium (DMEM 5671;
Sigma-Aldrich), with 10% fetal bovine serum (FBS, F7524; Sigma-Aldrich), 1%
HEPES (H3375; Sigma-Aldrich), 1% sodium pyruvate (P2256; Sigma-Aldrich), 1%
L-glutamine (G7513; Sigma-Aldrich) and 1% penicillin-streptomycin (P0781;
Sigma-Aldrich).
cultured and expanded in Dulbecco's modified Eagle's medium (DMEM 5671;
Sigma-Aldrich), with 10% fetal bovine serum (FBS, F7524; Sigma-Aldrich), 1%
HEPES (H3375; Sigma-Aldrich), 1% sodium pyruvate (P2256; Sigma-Aldrich), 1%
L-glutamine (G7513; Sigma-Aldrich) and 1% penicillin-streptomycin (P0781;
Sigma-Aldrich).
4
Isolation and Characterization of Human Nasal Cells
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All recruited participants had a microbiological swab taken from their middle meatus for culture, a cytological brushing for the generation of epithelial cells and a tissue biopsy for (i) the generation of fibroblast cells and (ii) tissue sections for histological analysis. Epithelial cells were grown from the brushings to near confluence at passage zero (P0), without trypsinisation or splitting of the cells. The cells were grown in submerged tissue culture conditions with Lonza BEGM media supplemented with 100 iu/ml penicillin/streptomycin. Fibroblast cells were cultured using an outgrowth technique from tissue biopsies to P1 in Sigma DMEM 5671supplemented with 100 iu/ml penicillin/ streptomycin, 10% fetal calf serum, 2mM L-Glutamine and 5ml Amphotericin B. All cells were cultured in standard sterile tissue culture conditions. Cultured cells were characterized and confirmed to be epithelial or fibroblast by immunocytochemistry using a panel of epithelial and mesenchymal markers (18) .
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