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Eagle s minimum essential medium alpha modification

Manufactured by Merck Group
Sourced in Germany

Eagle's minimum essential medium alpha modification is a cell culture medium used for the growth and maintenance of a variety of cell types. It provides the necessary nutrients and components to support cell proliferation and survival in in vitro cell culture systems.

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3 protocols using eagle s minimum essential medium alpha modification

1

Isolation and Culture of Gingival Mesenchymal Stem Cells

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G-MSCs were obtained from the healthy gingival collars around partially impacted third molars at the Christian-Albrechts University of Kiel, Germany. The study was performed in accordance with Helsinki Declaration revised in 2008 and approved by the Ethics Committee of Christian-Albrechts University of Kiel, Germany. Cells' isolation and culture were done as formerly described [14 (link), 15 (link)]. Briefly, gingival soft tissue collars were detached, deepithelized, cut into pieces, and washed in a basic medium, consisting of Eagle's minimum essential medium alpha modification (Sigma-Aldrich GmbH, Hamburg, Germany), with 15% fetal calf serum (FCS, HyClone, Logan, UT, USA), 400 mmol/ml L-glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin, and 1% amphotericin (all from Biochrom, Berlin, Germany). The soft tissue parts were left to adhere for 30 min in dry tilted culture flasks, before the basic medium was slowly added to them. The flasks were incubated at 37°C with 5% CO2 to let the cells grow and reach 80% confluence. The basic medium was replaced three times/week.
Immunomagnetic cell sorting employing anti-STRO-1 antibody (BioLegend, San Diego, CA, USA) and anti-IgM MicroBeads (Miltenyi Biotec, Bergisch Gladbach, Germany) were used to isolate the G-MSCs as described before [14 (link)]. The obtained G-MSCs were cultured in a basic medium.
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2

Mesenchymal Stem Cell Inflammatory Modulation

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Second passage G-MSCs were cultured in basic medium, consisting of Eagle’s minimum essential medium alpha modification (Sigma-Aldrich GmbH, Hamburg, Germany) supplemented with antibiotics (100 U mL−1 penicillin, 100 µg mL−1 streptomycin) and 1% amphotericin (all from Biochrom, Berlin, Germany) (control group), in basic medium, with 1 ng/mL IL-1β, 10 ng/mL TNF-α and 100 ng/mL IFN-γ (Pepro Tech Inc., Rocky Hill, NJ, USA) [5 (link),6 (link),8 (link),23 (link),24 (link),25 (link)] (inflammatory group), in basic medium with 250 µg/mL AA [21 (link)] and 20 µmol/L retinol [26 (link)] (AA/retinol group), or in inflammatory medium with 250 µg/mL AA and 20 µmol/L retinol (inflammatory/AA/retinol group). Media were exchanged three times per week.
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3

Fibroblast Cytocompatibility Assay with WPI and GTE

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Human fibroblasts were isolated from the gingiva of seven patients (4 female, aged 21, 25, 20 and 20 years and 3 male, aged 36, 21 and 21 years) cultured in cell culture medium (Eagle's minimum essential Medium Alpha modification (Sigma-Aldrich GmbH, Hamburg, Germany), antibiotics (100 U x mL-1 Penicillin, 100 µg x mL-1 Streptomycin, Biochrom, Berlin, Germany), 1 % Amphotericin (Biochrom), 15 % fetal bovine serum (FBS) (HyClone, Logan, Utah, USA) and 400 mmol x ml-1 L-Glutamine) for 7 days, then medium was exchanged three times per week Ethical approval (number D444/10) was obtained from University Hospital Schleswig-Holstein, Campus Kiel, Germany. Fibroblasts in the third passage were pooled and 5x10 3 were seeded in 96well plates, i.e. 1.56 x 10 4 cells/cm 2 . Cytocompatibility was assessed by incubating cells with medium containing 0, 50 or 800 µg/ml WPI and 0, 64, 128, 256, 512, 1024, 2048 or 4096 µg/m l GTE. Morphology was assessed by light microscopy after 7 days of culture. Medium was changed 3 times during this period.
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