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Mem vit

Manufactured by Thermo Fisher Scientific
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MEM-Vit is a specialized cell culture media formulation containing a balance of amino acids, vitamins, and other essential nutrients to support the growth and maintenance of a variety of cell types in vitro. It is designed to provide the necessary components for cell proliferation and survival without additional supplementation.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Dispase 2, by Merck Group (2 mentions) Collagenase xi, by Merck Group (2 mentions) Dispase, by Thermo Fisher Scientific (2 mentions) Poly l lysine (pll), by Merck Group (2 mentions) Collagenase, by Merck Group (1 mentions) Hank s buffered saline solution, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Nerve growth factor (ngf), by Merck Group (1 mentions) Pen strep, by Thermo Fisher Scientific (1 mentions) Collagenase type xi, by Merck Group (1 mentions) Hbss solution, by Thermo Fisher Scientific (1 mentions) Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions) Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions)

4 protocols using mem vit

1

Isolation of Murine Dorsal Root Ganglion Neurons

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Dorsal root ganglion (DRG) neurons from mice were dissociated as previously described.48 (link) Briefly, mice of either sex were anaesthetized with isoflourane and killed by cervical dislocation. The spinal cord was removed and 25–30 DRGs were extracted and washed in cold Hank’s buffered saline solution (Invitrogen). Ganglia were incubated in 900 U/ml collagenase (type XI Sigma) and 5.46 U/ml dispase (Invitrogen) for 45 min at 37°C in 5% CO2. After that, they were gently triturated using a flame-polished glass pipette in calcium free culture medium Hank’s buffered saline solution (Invitrogen) + 1% MEM-Vit (Invitrogen) + 10% FBS (Invitrogen) + 100 mg/ml penicillin/streptomycin (Invitrogen) and the resulting suspension was centrifuged. The pellet obtained was resuspended in culture medium with the following composition: MEM (Invitrogen) + 1% MEM-Vit (Invitrogen) + 10% FBS (Invitrogen) + 100 mg/ml penicillin/streptomycin (Invitrogen). Cells were plated on round glass coverslips treated with 0.01% poly-L-lysine (Sigma) and kept at 37°C, 5% CO2.
Marcotti A., Fernández-Trillo J., González A., Vizcaíno-Escoto M., Ros-Arlanzón P., Romero L., Vela J.M., Gomis A., Viana F, & de la Peña E. (2022). TRPA1 modulation by Sigma-1 receptor prevents oxaliplatin-induced painful peripheral neuropathy. Brain, 146(2), 475-491.
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2

Mouse Dorsal Root Ganglion Neuron Isolation

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Mouse DRG neurons were extracted and dissociated as previously described [55 (link)]. Briefly, the mice were euthanized by cervical dislocation. The spinal cord was removed and 20–40 DRGs were dissected and washed in cold HBSS solution (Thermo Fisher Scientific). For IHC, whole ganglia were directly fixed for 2 h in 4 % PFA. For ICC, ganglia were then incubated in 900 U/mL type XI collagenase (Sigma-Aldrich) and 5.46 U/mL dispase (Thermo Fisher Scientific) for 45 min at 37 °C in 5% CO2. After enzymatic treatment, the ganglia were mechanically dissociated using fire-polished glass pipettes in calcium-free solution containing HBSS (Thermo Fisher Scientific), 1% MEM-Vit (Thermo Fisher Scientific), 10% fetal bovine serum (Thermo Fisher Scientific) and 100 mg/mL penicillin/streptomycin. The cell suspension was centrifuged, and the pellet was resuspended in culture medium containing: MEM (Thermo Fisher Scientific), 1% MEM-Vit (Thermo Fisher Scientific), 10% fetal bovine serum (Thermo Fisher Scientific) and 100 mg/mL penicillin/streptomycin. The cells were then seeded onto 6 mm diameter glass coverslips previously coated with 0.01% poly-L-lysine (Sigma-Aldrich, St. Louis). Twenty-four hours after seeding, the cells were subjected to calcium imaging or fixed for 10 min in 4%PFA.
Hernández-Ortego P., Torres-Montero R., de la Peña E., Viana F, & Fernández-Trillo J. (2022). Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression. International Journal of Molecular Sciences, 23(24), 16164.
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3

Isolation and Culture of Nociceptive Neurons

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Nociceptive neurons were cultured as described previously [47 (link)]. Briefly, TG and DRG were dissected out from mice or rat of 2–4 month old and incubated with collagenase XI (0.66 mg/mL) and dispase II (3 mg/mL) (Sigma-Aldrich) in an INC-mix solution (NaCl 155 mM; K2HPO4 1.5 mM; HEPES 10 mM; glucose 5 mM; at pH 7,4). Enzymatic digestion was performed for 45 min at 37°C in 5% CO2, and cells were cultured in minimum essential media (MEM) supplemented with 10% FBS, Pen/Strep 100 mg/mL, MEM-vit (Invitrogen, Carlsbad CA). Cells were plated on 12-mm poly-L-lysine-coated glass coverslips and cultured for 2 days. Animal experiments were conducted in accordance with the principles and procedures of the National Institutes of Health Guidelines for the Care and Use of Laboratory and the Bioethical Committee of Universidad Católica del Norte, Coquimbo, and the Ethics Committee of the Biology Department, Faculty of Sciences, Universidad of Chile, Santiago, Chile.
Coddou C., Sandoval R., Castro P., Lazcano P., Hevia M.J., Rokic M., Hall B., Terse A., Gonzalez-Billault C., Kulkarni A.B., Stojilkovic S.S, & Utreras E. (2017). Cyclin-dependent kinase 5 modulates the P2X2a receptor channel gating through phosphorylation of C-terminal threonine 372. Pain, 158(11), 2155-2168.
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4

Culturing Mouse Trigeminal Neurons

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Trigeminal neurons were cultured as described previously [28 (link)]. Briefly, mouse TG were dissected out and incubated with Collagenase XI (0,66 mg/mL) and Dispase II (3 mg/mL) (Sigma-Aldrich, San Louis, MO) in a INC-mix solution (NaCl 155 mM; K2HPO4 1.5 mM; HEPES 10 mM; glucose 5 mM; at pH 7,4). The enzymatic digestion was performed for 45 min at 37°C in 5% CO2, and cells were cultured in Minimum Essential Media (MEM) supplemented with 10% fetal bovine serum (FBS), Pen/Strep 100 µg/mL, MEM-vit (Invitrogen, Carlsbad, CA), and nerve growth factor (100 ng/ml; Sigma-Aldrich, San Louis, MO). Cells were plated on 6 or 13 mm poly-l-lysine-coated glass coverslips and used after 4 h for Ca2+ imaging analysis or after 18 h for immunofluorescence analysis. In some cases, roscovitine (10 µM) was added for 4 h before Ca2+ imaging experiments.
Rozas P., Lazcano P., Piña R., Cho A., Terse A., Pertusa M., Madrid R., Gonzalez-Billault C., Kulkarni A.B, & Utreras E. (2016). Targeted overexpression of Tumor Necrosis Factor-α increases Cyclin-dependent kinase 5 activity and TRPV1-dependent Ca2+ influx in trigeminal neurons. Pain, 157(6), 1346-1362.
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