P. gingivalis strain W83 was purchased from the American Type Culture Collection (ATCC BAA-308™, LGC Standards GmbH, Wesel, Germany) and grown at 37 °C in brain-heart-infusion broth (Difco, BD, Heidelberg, Germany) with hemine (5 μg/ml) and menadione (1 μg/ml) (Sigma-Aldrich, Munich, Germany) under anaerobic conditions using the Anaerocult A System (Merck, Darmstadt, Germany).
Hemine
Manufactured by Merck Group
Sourced in Germany
Hemine is a laboratory equipment product manufactured by the Merck Group. It is a specialized tool designed for specific functions in a laboratory setting. The core function of Hemine is to provide an accurate and reliable solution for [core function]. Further details on the intended use of this product are not available.
Automatically generated - may contain errors
Lab products found in correlation
Menadione, by Merck Group (3 mentions)
Anaerocult a system, by Merck Group (1 mentions)
Anaerogen system, by Thermo Fisher Scientific (1 mentions)
Porphyromonas gingivalis w83, by American Type Culture Collection (1 mentions)
Ferric citrate, by Merck Group (1 mentions)
Yeast extract, by Merck Group (1 mentions)
N acetylmuramic acid, by Merck Group (1 mentions)
4 protocols using hemine
1
Culturing Porphyromonas gingivalis W83
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P. gingivalis strain W83 was purchased from the American Type Culture Collection (ATCC BAA-308™, LGC Standards GmbH, Wesel, Germany) and grown at 37 °C in brain-heart-infusion broth (Difco, BD, Heidelberg, Germany) with hemine (5 μg/ml) and menadione (1 μg/ml) (Sigma-Aldrich, Munich, Germany) under anaerobic conditions using the Anaerocult A System (Merck, Darmstadt, Germany).
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P. gingivalis strain W83 was purchased from the American Type Culture Collection (ATCC BAA-308™, LGC Standards GmbH, Wesel, Germany) and grown at 37 °C in brain-heart-infusion broth (Difco, BD, Heidelberg, Germany) with hemine (5 μg/ml) and menadione (1 μg/ml) (Sigma-Aldrich, Munich, Germany) under anaerobic conditions using the Anaerocult A System (Merck, Darmstadt, Germany).
2
Cultivation of Porphyromonas gingivalis
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Porphyromonas gingivalis W83 was purchased from the American Type Culture Collection (ATCC, LGC Standards GmbH, Wesel, Germany). It was grown at 37°C in brain-heart-infusion broth (Difco, BD, Heidelberg, Germany) with hemine (5 µg/ml) and menadione (1 µg/ml) (Sigma-Aldrich, Munich, Germany) under anaerobic conditions using the Anaerogen system (ThermoFisher, Dreieich, Germany) until late log phase (OD=1).
3
Optimized Pregnancy-Friendly AIN-76A Diet
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A purified powder-based AIN-76A diet was optimized to maintain iron bioavailability (calcium level limited to 0.09%) while taking care to meet pregnancy requirements in terms of protein and amino-acids (UE 1298, Sciences de l’Animal & de l’Aliment, INRAE). In order to prepare frozen portions that were daily distributed into cage feeders and thus, limit diet lipid peroxidation, the powdered form was further transformed into jellied mass cut into cubes by incorporating porcine gelatin (8%, Louis François) and completed with oil (5% safflower oil, Fischer scientific) and 1,4 μmol iron/g diet (0,094% hemine or 0,0036% ferric citrate, Sigma) before storage under vacuum at −20 °C.
4
Blood Agar Enrichment Protocol
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Blood agar enriched with hemine, menadione, and N-acetylmuramic acid was prepared. Medium composition comprised 5% defibrinated sheep blood (NewProv LTDA, Pinhais, PR, Brasil), 2.6% brain heart infusion agar (BHI—Difco Laboratories, Detroit, MI, USA), 2% trypticase soy agar (TSA—Difco (Franklin Lakes, NJ, USA)), 1% yeast extract (BBL, Cockeysville, MD, USA), 1% hemine (Sigma Chemical Co., St. Louis, MO, USA), 1% N-acetylmuramic acid (Sigma Chemical Co.), and 0.5% menadione (Sigma Chemical Co.). Agar mixture was sterilized in autoclave and, after cooling it to 50 °C, sheep blood, menadione, and N-acetylmuramic acid were added to the mixture under aseptic conditions. Medium was then dispensed into 96-well plates (TPP, Zellkultur testplatte, Trasadingen, Switzerland), 150 µL per well, and allowed to solidify. Plates were stored at 4 °C.
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