An immortalized mouse hippocampal cell line, HT-22, was cultured in Hyclone Dulbecco’s modified Eagle’s medium (DMEM)/high glucose (Fisher Scientific, Waltham, MA, USA) with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) and 1% penicillin/streptomycin (Fisher Scientific). The HT-22 mouse hippocampal neuronal cell line was provided by The Salk Institute for Biological Research (La Jolla, CA, USA), and cell passages 15–23 were used. When HT-22 cells were at least 80% confluent, the cells were trypsinized and spun down at 325.5 × g for 3 min. The cells were counted with Nexcelom Bioscience Cellometer AutoT4 (Lawrence, MA, USA). Primary mouse cortical neurons were purchased from Life Technologies (Carlsbad, CA, USA) and cultured in Neurobasal® Medium supplemented with GlutaMAX™ – I and B-27® supplement (Life Technologies).
Hyclone dulbecco s modified eagle s medium dmem high glucose
Manufactured by Thermo Fisher Scientific
Sourced in United States
Hyclone Dulbecco's modified Eagle's medium (DMEM)/high glucose is a cell culture medium formulation. It is designed to support the growth and maintenance of a variety of cell types in an in vitro environment.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (3 mentions)
Cellometer auto t4, by Revvity (2 mentions)
Neurobasal medium, by Thermo Fisher Scientific (1 mentions)
Hyclone fetal bovine serum, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
4 protocols using hyclone dulbecco s modified eagle s medium dmem high glucose
1
In Vitro Hippocampal and Cortical Neuron Cultures
2
HT-22 Mouse Hippocampal Cell Culture
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The HT-22 immortalized mouse hippocampal cells were obtained from the Salk Institute for Biological Research (La Jolla, CA, USA). HT-22 cells were cultured in Hyclone Dulbecco’s modified Eagle’s medium (DMEM)/high glucose (Fisher Scientific, Waltham, MA, USA) with 10% fetal bovine serum (FBS; Atlanta Biologicals, Flowery Branch, GA, USA) and 1% penicillin/streptomycin (Fisher Scientific). When cells were at least 80% confluent, they were trypsinized and spun down at 325.5× g for 3 min. Cells were counted with a Nexcelom Bioscience Cellometer AutoT4 (Lawrence, MA, USA). Cell passages 5–18 were used for all experiments.
3
Culturing C6 Rat Glial Cells
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The C6 rat glial cell line was obtained from the American Type Culture Collection (ATCC #CCL 107). C6 cells were maintained in Hyclone Dulbecco’s modified Eagle’s medium (DMEM)/high glucose (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals) and 1% penicillin/streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). Once cells reached 80% confluency, they were trypsinized and spun down at 325.5 × g for 3 min. Cells were counted with a Nexcelom Bioscience Cellometer AutoT4 (Lawrence) and seeded at a density of 1 × 106 cell/well in 6-well cluster plates, incubated overnight at 37°C in 5% CO2.
4
Hypoxic Regulation of Breast Cancer Cells
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Human breast cancer cell lines MDA-MB-231 and MDA-MB-468 were obtained from the Cell Biology Institute of the Chinese Academy of Sciences. The cells were cultured in HyClone Dulbecco’s modified Eagle’s medium (DMEM) high glucose (Thermo Fisher Scientific) supplemented with 10% (v/v) HyClone fetal bovine serum (FBS) in a humidified incubator at 37 °C with 5% CO2. Cells were grown on plastic dishes for protein extraction and wound-healing assays. pEGFP-N1 vector containing a dominant negative Rac1-T17 N insert was provided by Dr. Shoshana Ravid of the Hebrew University in Jerusalem, Israel. Cells were transfected with either pEGFP-N1 or pEGFP-N1 expressing Rac1-T17 N using Lipofectamine 2000 per the manufacturer’s instructions (Invitrogen).
Hypoxic conditions were maintained by exposing cells to a continuous flow of a humidified mixture of 1% O2, 5% CO2 and 94% N2 at 37 °C for the indicated time.
Hypoxic conditions were maintained by exposing cells to a continuous flow of a humidified mixture of 1% O2, 5% CO2 and 94% N2 at 37 °C for the indicated time.
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