Human T84 IECs were grown in Dulbecco’s modified Eagle’s medium (DMEM) -F12 50:50 (Sigma) with supplements as previously described.57 (link) CHO-K1 and SW480 cells (used for transfection and TopFlash assays) were passaged in DMEM containing high glucose and supplemented with 10% FBS, 100 units/ml penicillin, 100 μg/ml streptomycin, 10 mM Hepes (pH 7.4), 2 mM L-glutamine, and 1% non-essential amino acids. PMNs were isolated from human blood that was drawn from healthy volunteers and handled according to protocols for the protection of human subjects, as approved by the Emory University Hospital Institutional Review Board. PMNs were isolated by density gradient centrifugation57 (link), 58 (link) and were used in experiments within 2 h of isolation.
Dmem f12 50 50
Manufactured by Merck Group
Sourced in United States
DMEM/F-12 50:50 is a cell culture medium that provides a balanced salt solution to support the growth and maintenance of a variety of cell types. It is a 1:1 mixture of Dulbecco's Modified Eagle's Medium (DMEM) and Ham's F-12 Nutrient Mixture. This medium is commonly used in research and biomedical applications.
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Lab products found in correlation
3 protocols using dmem f12 50 50
1
Cell Culture and PMN Isolation
2
Isolation and Characterization of Mesenchymal Stem Cells
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As done previously [51 (link),53 ], bone marrow mononuclear cells were separated by centrifugation in a Ficoll–Hypaque gradient (density 1.077 g/cm2; Sigma, St Louis, MO, USA), washed twice with Hanks’ balanced saline solution (HBSS), and suspended in Dulbecco’s minimum essential medium-F12 50/50 (DMEM-F12 50/50, Sigma, St Louis, MO, USA) containing 10% fetal calf serum (FCS; Gibco BRL, Grand Island, NY, USA). Hematopoietic cells and non-adherent cells were removed by washing the culture the next day, followed with changes in medium. Adherent cells after the second subculture, referred to as second generation MSCs, were used to prepare frozen stocks.
Chondrogenic, adipogenic, and osteogenic differentiation assays were performed to ensure that the cells had the characteristics of MSCs. These samples were used to initiate the MSC culture for in vivo studies.
Chondrogenic, adipogenic, and osteogenic differentiation assays were performed to ensure that the cells had the characteristics of MSCs. These samples were used to initiate the MSC culture for in vivo studies.
3
Cell Culture and PMN Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human T84 IECs were grown in Dulbecco’s modified Eagle’s medium (DMEM) -F12 50:50 (Sigma) with supplements as previously described.57 (link) CHO-K1 and SW480 cells (used for transfection and TopFlash assays) were passaged in DMEM containing high glucose and supplemented with 10% FBS, 100 units/ml penicillin, 100 μg/ml streptomycin, 10 mM Hepes (pH 7.4), 2 mM L-glutamine, and 1% non-essential amino acids. PMNs were isolated from human blood that was drawn from healthy volunteers and handled according to protocols for the protection of human subjects, as approved by the Emory University Hospital Institutional Review Board. PMNs were isolated by density gradient centrifugation57 (link), 58 (link) and were used in experiments within 2 h of isolation.
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