The hippocampal neurons (HT22) cell line, neuroblastoma (N2A) cell line, and human neuroblastoma (SH-SY5Y) cells were purchased from ATCC Company (Manassas, VA, USA) with short-tandem repeat profiling for authentication. The cells were grown in DMEM (Dulbecco’s modified Eagle medium, Gibco Invitrogen) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin. The cells were maintained in a 5% CO2 culture incubator at 37 °C.
N2a cell line
Manufactured by American Type Culture Collection
Sourced in United States
The N2a cell line is a neuroblastoma cell line derived from the brain of a mouse. It is a commonly used model for studying neurological processes and functions. The N2a cell line can be used for a variety of cell-based assays and experiments.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Sh sy5y, by American Type Culture Collection (1 mentions)
Ara c, by Merck Group (1 mentions)
Sh sy5y cell line, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Fs201 02, by Transgene (1 mentions)
Hanks balanced salt solution (hbss), by Beyotime (1 mentions)
Dmem f12 medium, by Cytiva (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
U87mg luc cell line, by Shanghai Model Organisms (1 mentions)
U87mg cell line, by American Type Culture Collection (1 mentions)
Bv2 cell line, by American Type Culture Collection (1 mentions)
U251 cell line, by American Type Culture Collection (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
10 protocols using n2a cell line
1
Cell Line Cultivation and Maintenance
2
Culturing Mouse Neuroblastoma and Primary Visual Cortical Neurons
3
Isolation and Culture of Mouse and Human Neuronal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse neural crest-derived N2A cell line and human neuroblastoma SH-SY5Y cell line were purchased from ATCC and cultured in DMEM/F12 medium (containing 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 0.25 μg/ml amphotericin B). Medium was changed every 2 days. For culturing mouse primary neuronal cells, cortical neurons were isolated from the cortex of postnatal day 1 mice. Dissected tissue was treated with 0.0625 mg/ml trypsin and 0.0625 mg/ml DNasein 1 × HBSS buffer without calcium and magnesium for 10 min at 37 °C. Cells were washed once with the tissue culture medium, centrifuged at 1500 × g for 3 min, and then placed for initial growth in a 50% glial-conditioned medium (containing 0.25% glucose, 2 mM glutamate, 10% FCS, 500 nm insulin, 1 × vitamin mixture, and 1% antibiotic-antimycotic). The cells were cultured in neurobasal/B27 medium.
4
Cholinergic Hybrid Cell Line Maintenance and Differentiation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SN56 cells were maintained Dulbecco’s minimal Eagle’s medium (DMEM, Invitrogen) supplemented with 10% fetal bovine serum and 50ug/ml of penicillin/streptomycin, in an incubator at 37°C with 5% CO2. Cells were split every 3–4 days; depending on confluency. SN56 cells are a cholinergic hybrid cell line, which were generated by fusing dissociated mouse septal neurons and N18TG2 neuroblastoma cells. They were chosen for our experiments because after differentiation they are cholinergic, possess neuronal morphology, and will express APP [29 (link)–31 (link)]. These were provided to us by Dr. Jane Rylett (Western University, London Ontario, Canada). In some experiments, we also examine trafficking in the N2a cell line (ATCC, Manassas, Virginia, USA). For microscopy, cells were seeded on glass-bottomed culture dishes (MatTek) one day before transfection. Cells were transfected using Lipofectamine 2000 (Invitrogen) following manufactures directions. After a 24hrs, cells were differentiated in DMEM using 1mM dibutyrl cyclic AMP (dbcAMP; Sigma) and imaged or fixed.
5
Starvation of 293T and N2a cell lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human embryonic kidney cell line 293T and mouse neuroblastoma N2a cell line were originally obtained from the American Type Culture Collection (ATCC). The cell line was maintained in Dulbecco’s modified Eagle’s medium (DMEM, 11,665,092, Gibco). All media were supplemented with 10% FBS (FS201-02, Transgene Biotech) and 1% penicillin and streptomycin. Cells were incubated at 37 °C in a humid 5% CO2:95% air environment.
For starvation, after aspiration of the cell culture medium and two washes with PBS, HBSS (C0219, Beyotime) medium was added and then incubated for 6 h.
For starvation, after aspiration of the cell culture medium and two washes with PBS, HBSS (C0219, Beyotime) medium was added and then incubated for 6 h.
6
Transfection of N2a cells with APP constructs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Neuro 2A (N2a) cell line was obtained from the American Type Culture Collection (ATCC, #CCL-131). N2a cells were cultured in DMEM/F12 medium (Cytiva) containing 10% FCS, 15 mM HEPES, 2.5 mM L-glutamine and 0.1 mg/ml penicillin and streptomycin. Transfection was performed using Lipofectamine 2000 according to the manufacturer’s instructions (Invitrogen). In brief, N2a cells were seeded at 1 × 105 cells/well in 24-well plates, and transfected 24 h later with the indicated DNA plasmid (0.5 μg/well) using Lipofectamine 2000 transfection reagent (1 μl/well). pcDNA FRT TO- APP695 and pcDNA FRT TO- APPSwed/Ind were gifts from Aleksandra Radenovic (Addgene plasmids #114193 and #114194, respectively). pcDNA3.3 plasmid (Themo Fisher Science, #K830001) was obtained from Invitrogen, and was used as a negative control (Empty).
7
In Vitro OGD/R Injury Model and Dexmedetomidine Treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mouse N2A cell line was purchased from American Type Culture Collection
(Manassas, VA, USA). Cells were cultured in DMEM medium (Gibco BRL, Life
Technologies Inc., Gaithersburg, MD, USA) supplemented with 10% fetal bovine
serum (Gibco BRL, Life Technologies Inc.). The culture plates were incubated at
37 °C in a humidified atmosphere containing 5% CO2. In order to
generate the in vitro OGD/R injury model as previously described,14 (link) N2A cells were cultured in serum/glucose-free DMEM medium in a humidified
atmosphere containing 5% CO2 and 95% N2 at 37 °C for 4 h,
followed by their return to DMEM supplemented with 10% fetal bovine serum for a
12-h recovery in normoxic conditions. Then, Non-OGD or OGD/R N2A cells were
treated with dexmedetomidine solutions (Abbott Laboratories, Worcester, MA, USA)
at 50 ng/ml, 100 ng/ml and 500 ng/ml for 60 min at 37 °C for subsequent
experiments. In addition, for p38 microtubule associated protein
kinase/extracellular signal-regulated kinases (MAPK/ERK) signalling inhibition,
cells were treated with the inhibitor CV-65 (Abcam®, Cambridge, MA, USA) at 20
μM for 60 min at 37 °C as previously described.15 (link)
(Manassas, VA, USA). Cells were cultured in DMEM medium (Gibco BRL, Life
Technologies Inc., Gaithersburg, MD, USA) supplemented with 10% fetal bovine
serum (Gibco BRL, Life Technologies Inc.). The culture plates were incubated at
37 °C in a humidified atmosphere containing 5% CO2. In order to
generate the in vitro OGD/R injury model as previously described,14 (link) N2A cells were cultured in serum/glucose-free DMEM medium in a humidified
atmosphere containing 5% CO2 and 95% N2 at 37 °C for 4 h,
followed by their return to DMEM supplemented with 10% fetal bovine serum for a
12-h recovery in normoxic conditions. Then, Non-OGD or OGD/R N2A cells were
treated with dexmedetomidine solutions (Abbott Laboratories, Worcester, MA, USA)
at 50 ng/ml, 100 ng/ml and 500 ng/ml for 60 min at 37 °C for subsequent
experiments. In addition, for p38 microtubule associated protein
kinase/extracellular signal-regulated kinases (MAPK/ERK) signalling inhibition,
cells were treated with the inhibitor CV-65 (Abcam®, Cambridge, MA, USA) at 20
μM for 60 min at 37 °C as previously described.15 (link)
8
Culture Conditions for Brain Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human GBM U87MG cell line, human brain endothelial hCMEC/D3 cell line, N2a cell line, BV2 cell line, U251 cell line, U251TR cell line were purchased from the the American Type Culture Collection (ATCC). U87MG-Luc cell line was purchased from Shanghai Model Organisms Center (Shanghai, China). The above cell lines were cultured in DMEM medium supplemented with 10% (V/V) fetal bovine serum and 1% (V/V) penicillin and streptomycin. The X01 glioblastoma stem cells (GSCs) were kindly provided by Professor Jong Bae Park from the National Cancer Center of South Korea, the more details are in the previous works49 (link),61 (link)–64 (link). X01 cells were maintained in DME/F12 supplemented with epidermal growth factor (EGF, 10 ng ml−1), basic fibroblast growth factor (bFGF, 10 ng ml−1), B27, and 1% penicillin and streptomycin. These cells were cultured as a monolayer in a humidified atmosphere containing 5% CO2 at 37 °C.
9
Culturing mHippoE-18 and N2a Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mHippoE-18 cell line (embryonic mouse hippocampal cell line) was purchased from Cederlane (Burlington, Ontario, Canada), whereas the N2a cell line (mouse neuroblastoma cell line) was purchased from the American Type Culture Collection (ATCC, VA, USA) (CCL-131). Cells were cultured in DMEM medium supplemented with 5% fetal bovine serum and incubated at 37 °C in an atmosphere of 5% CO2. They were split for subcultures every 2–3 days.
10
In Vitro I/R Injury Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The N2a cell line was purchased from the American Type Culture Collection (ATCC, Rockville, MD, United States). Dulbecco’s modified Eagle’s medium (DMEM)-F12 medium (Thermo Fisher Scientific, Shanghai, China) containing 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin were used for cell culture. When the cells were 80% confluent, 0.25% trypsin (Thermo Fisher HyClone, Utah, United States) was used for cell trypsinization and passage (Bao et al., 2018 (link)). Cells in the logarithmic growth phase were selected for later use. To stimulate I/R injury in vitro, cells were subjected to 3 h of hypoxia and 3 h of reoxygenation, according to a previous study (Zhao et al., 2018 (link)).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!