MEM (Eagle) Liquid Medium

Mammalian cell culture media HOb

Experiment
Mammalian cell culture media HOb
Product
MEM (Eagle) Liquid Medium from Merck Millipore
Manufacturer
Merck Millipore

Protocol tips

Protocol tips
The human osteoblasts were cultivated in modified Eagle’s osteogenic cell culture medium (MEM; Biochrom AG, Berlin, Germany) containing 10% fetal calf serum (FCS), 1% penicillin/streptomycin, 1% amphotericin B, and 1% HEPES buffer (all from Gibco-Invitrogen, Darmstadt, Germany) without calcium, and the osteogenic additives dexamethasone (100 mM), L-ascorbic acid (50 μg/mL), and β-glycerophosphate (10 mM) (all from Sigma-Aldrich, Munich, Germany).The osteogenic differentiation of the human primary osteoblasts was confirmed by immunohistochemical detection of the enzyme alkaline phosphatase using a fuchsin+substrate chromogen (DAKO, Hamburg, Germany). For the further tests, the isolated cells were cultured in 25-cm2 flasks with 8 ml of osteoblasts in the same medium but without the 1% penicillin/streptomycin and under standard cell culture conditions (5% CO2 and 37°C).

Publication protocol

The human primary osteoblasts were taken from the spongiosa of the femoral heads of patients who underwent total hip replacement. The study was approved by the Local Ethical Committee of Rostock, Germany (registration number: A2010-10), and an informed consent was signed by each patient.

The human osteoblasts were cultivated in modified Eagle’s osteogenic cell culture medium (MEM; Biochrom AG, Berlin, Germany) containing 10% fetal calf serum (FCS), 1% penicillin/streptomycin, 1% amphotericin B, and 1% HEPES buffer (all from Gibco-Invitrogen, Darmstadt, Germany) without calcium, and the osteogenic additives dexamethasone (100 mM), L-ascorbic acid (50 μg/mL), and β-glycerophosphate (10 mM) (all from Sigma-Aldrich, Munich, Germany). The osteogenic differentiation of the human primary osteoblasts was confirmed by immunohistochemical detection of the enzyme alkaline phosphatase using a fuchsin+substrate chromogen (DAKO, Hamburg, Germany). For the further tests, the isolated cells were cultured in 25-cm2 flasks with 8 ml of osteoblasts in the same medium but without the 1% penicillin/streptomycin and under standard cell culture conditions (5% CO2 and 37°C).

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Papers

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Paper title
Co-Culture of . and Human Osteoblasts on Implant Surfaces: An Advanced Model for Implant-Associated Infections
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Manufacturer protocol

Download the product protocol from Merck Millipore for MEM (Eagle) Liquid Medium below.

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