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Di tnc1

Manufactured by American Type Culture Collection
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The DI TNC1 is a laboratory instrument designed for automated thawing and dispersion of frozen cell culture samples. It provides a controlled and consistent process to efficiently prepare cells for further analysis or experimentation.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Trypsin, by Atlanta Biologicals (2 mentions) Dulbecco phosphate buffered saline (dpbs), by Atlanta Biologicals (2 mentions) Raw264, by Thermo Fisher Scientific (1 mentions) Dmem high glucose medium, by Thermo Fisher Scientific (1 mentions) Raw264, by American Type Culture Collection (1 mentions) Tib 152, by American Type Culture Collection (1 mentions) Hek293, by American Type Culture Collection (1 mentions) Lipofectamine 3000 transfection kit, by Thermo Fisher Scientific (1 mentions) Hek293t, by American Type Culture Collection (1 mentions) Ctx tna2, by American Type Culture Collection (1 mentions)

9 protocols using di tnc1

1

Macrophage and Astrocyte Cell Lines

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Cell lines were purchased from the American Type Culture Collection. The murine macrophage cell line RAW 264.7 (ATCC® TIB-71™) was selected for efficacy studies, whereas rat astrocyte DITNC1 (ATCC® CRL-2005™) was used for toxicity studies.
Cells were grown in DMEM high glucose medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS—Thermo Fisher Scientific) and 1% penicillin/streptomycin (100 U mL−1/100 µg mL−1) (Thermo Fisher Scientific) at 37 °C in a humidified incubator of 5% CO2.
For efficacy studies, RAW 264.7 cells were seeded on a 24-well plate at a density of 12.000 cells/well and treated after 3 days in culture with conditioned medium, prepared as described below.
For GLP toxicity studies, DITNC1 cells were validated according to the doubling time and absence of mycoplasma contamination. Soluble molecules and conditioned medium for toxicity studies were used as described in the dedicated sections.
Dolci L.S., Perone R.C., Di Gesù R., Kurakula M., Gualandi C., Zironi E., Gazzotti T., Tondo M.T., Pagliuca G., Gostynska N., Baldassarro V.A., Cescatti M., Giardino L., Focarete M.L., Calzà L., Passerini N, & Bolognesi M.L. (2021). Design and In Vitro Study of a Dual Drug-Loaded Delivery System Produced by Electrospinning for the Treatment of Acute Injuries of the Central Nervous System. Pharmaceutics, 13(6), 848.
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2

Culturing Rat Astrocytes for Experiments

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DI TNC1 (ATCC, Manassas, VA, USA) rat astrocytes were cultivated in Dulbecco’s modified Eagle medium (DMEM) containing 10% fetal bovine serum, 2% glutamine, 1% sodium pyruvate, and 1% penicillin–streptomycin. Cultures were maintained at 37 °C with 5% CO2 until they reached the 80% confluence necessary to perform the experiments.
Coelho R., De Benedictis C.A., Sauer A.K., Figueira A.J., Faustino H., Grabrucker A.M, & Gomes C.M. (2024). Secondary Modification of S100B Influences Anti Amyloid-β Aggregation Activity and Alzheimer’s Disease Pathology. International Journal of Molecular Sciences, 25(3), 1787.
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3

Culturing Rat Astrocytes and Glioblastoma Cells

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DI TNC1 (ATCC, Manassas, VA, USA: CRL-2005) rat astrocytes were seeded (50,000 cells/mL) on poly-L-lysine (0.1 mg/mL)-coated 10 cm Petri dish and 24 multiwell and incubated in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum, 2% glutamine, 1% sodium pyruvate and 1% penicillin–streptomycin for 3 days until they were confluent. The culture was incubated at 37 °C in a suitable incubator with 5% CO2.
C6 glioblastoma (ATCC, Manassas, VA, USA: CCL-107) rat glioblastoma cells were seeded (50,000 cells/mL) on poly-L-lysine (0.1 mg/mL)-coated 10 cm Petri dish and 24 multiwell and incubated in F-12K nutrient mixture (1X) containing 20% fetal bovine serum, 1% penicillin–streptomycin for 72 h until they reach the 80–85% confluence. The cells were incubated at 37 °C in a suitable incubator with 5% CO2 in an air atmosphere.
De Benedictis C.A., Haffke C., Hagmeyer S., Sauer A.K, & Grabrucker A.M. (2021). Expression Analysis of Zinc Transporters in Nervous Tissue Cells Reveals Neuronal and Synaptic Localization of ZIP4. International Journal of Molecular Sciences, 22(9), 4511.
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4

Validating Anti-SV2A Antibody in Cell Lines

Cited 1 time
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To validate the rabbit anti-SV2A antibody used for immunohistochemistry
(cat#TA322365; Origene, Rockville, MD, USA), we performed an immunoblot of
lysate from mouse brain, from an immortalized rat astrocyte cell line (DI TNC1;
catalogue #CRL-2005; ATCC, Gaithersburg, MD, USA) and from an immortalized human
T-lymphocyte cell line (TIB-152; ATCC), using standard methods as we described.40 (link)
Tsymbalyuk S., Smith M., Gore C., Tsymbalyuk O., Ivanova S., Sansur C., Gerzanich V, & Simard J.M. (2019). Brivaracetam attenuates pain behaviors in a murine model of neuropathic pain. Molecular Pain, 15, 1744806919886503.
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5

Cell Culture Protocols for Neuroscience

Cited 1 time
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HEK293 (ATCC CRL-1573) and DI TNC1 (ATCC CRL-2005) and BV2 cell lines were cultured in D10 (DMEM-10% FBS with 1X Pen/Strep). Schwann cells harvested from adult rat sciatic nerves [24 (link)] were seeded on poly-L-lysine (PLL) coated petri dishes in D10-3M media (D10, 2 μM Forskolin, 10 nM Heregulin, and 20 μg/mL of pituitary extract). Rat astrocytes isolated from E18-19 rat cortices [25 (link)] were cultured in astrocyte medium (A1261301, Gibco, Carlsbad, CA) and grown in 75 cm2 vented culture flasks. All cell cultures were maintained inside a humidified incubator at 37°C and 5% CO2.
Rao S., Morales A.A, & Pearse D.D. (2015). The Comparative Utility of Viromer RED and Lipofectamine for Transient Gene Introduction into Glial Cells. BioMed Research International, 2015, 458624.
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6

Culturing Diverse Rat Cell Lines

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N27 (rat neural cell) were obtained from EMD Millipore (Billercia, MA, USA) and DI TNC1 (rat astrocyte) and RG2 (rat glioma) cell lines were obtained from ATCC (Manassas, VA, USA). BV-2 (murine microglia) cells were a kind gift from Dr Hideyuki Takahashi from Yale Cellular Neuroscience department. N27 cells were cultured in RPMI 1640 media supplemented with 10% FBS and 1% pen/strep. DI TNC1, RG2 and BV-2 cell lines were cultured in 4.5 g l−1 glucose DMEM media supplemented with 10% FBS and 1% pen/strep.
Song E., Gaudin A., King A.R., Seo Y.E., Suh H.W., Deng Y., Cui J., Tietjen G.T., Huttner A, & Saltzman W.M. (2017). Surface chemistry governs cellular tropism of nanoparticles in the brain. Nature Communications, 8, 15322.
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7

Cytokine-Induced Astrocyte Modeling

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Both human embryonic kidney 293T (HEK293T) cells and rat astrocytes CTX-TNA2 and DI TNC1 were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) [15] (link), which were seeded in Dulbecco's modi ed Eagles Medium containing 10% fetal bovine serum (FBS), 4 mM glutamine, 100 µg/mL streptomycin, 100 U/mL penicillin and 4.5 g/L glucose under an atmosphere of 5% CO 2 . CTX-TNA2 and DI TNC1 cells with stable growth were inoculated into 24 well plates and treated with interleukin (IL)-1β (10 ng/h) for 24 h for subsequent experiments. pCDNA3.1(+) plasmid was purchased from Kelei Bio. (Shanghai, China). NEAT1 mimic, miR-139 mimic and si-ROCK1were purchased from Thermo Fisher Scienti c (Waltham, MA, USA). The plasmid transfection was performed with the Lipofectamine 3000 transfection Kit (Invitrogen Inc., Carlsbad, CA, USA), and the results were veri ed by reverse transcription quantitative polymerase chain reaction (RT-qPCR).
Li X., Hao F., Tao S., Wang W., Xiao X., Guo D., Ma H., & Liu X. (2020). Long non-coding RNA NEAT1 aggravates epilepsy and seizure-induced neuronal damage by regulating microRNA-139/ROCK1 axis.
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8

Neuroblastoma Cells and Astrocytes Protocol

Cited 4 times
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Mouse brain neuroblastoma cells (N2-A) and rat primary astrocytes (DI-TNC1) were purchased from American Type Culture Collection (ATCC, Manassas, VA). N2- A cell line used in the current investigation is a neuronal cell line known for its high lactate production compare to other cell lines. We, as well as others, have used this cell line and is considered an appropriate model to evaluate potential anti-cancer agents [22 (link), 23 (link)]. We also used the N2-A cell line to investigate the “Warburg Effect” phenomenon [24 (link)], and cancer cells metabolism [25 (link), 26 (link)]. On the other hand, the DI-TNC1 is an astrocyte immortal cell line with lower lactate efflux production compared to N2-A cells, an observation in our lab. The DI-TNC1 is very important in controlling brain energy metabolism [27 (link), 28 (link)]. Cell culture Dulbecco's Modified Eagle Medium (DMEM), fetal bovine serum (FBS), penicillin/streptomycin, DPBS, and trypsin were all from Atlanta Biologicals (Atlanta, GA, USA). Cells were cultured in 75-cm TC flask at 37°C in humidified 5% CO2 incubator and were subcultured as needed with trypsin/EDTA. Growing media was supplemented with 10% FBS (v/v), 4 mM L-glutamine, and 1% penicillin /streptomycin.
Messeha S.S., Zarmouh N.O., Taka E., Gendy S.G., Shokry G.R., Kolta M.G, & Soliman K.F. (2016). The Role of Monocarboxylate Transporters and Their Chaperone CD147 in Lactate Efflux Inhibition and the Anticancer Effects of Terminalia chebula in Neuroblastoma Cell Line N2-A. European journal of medicinal plants, 12(4), EJMP.23992.
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9

Neuroblastoma Cells and Astrocytes Protocol

Cited 4 times
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Mouse brain neuroblastoma cells (N2-A) and rat primary astrocytes (DI-TNC1) were purchased from American Type Culture Collection (ATCC, Manassas, VA). N2- A cell line used in the current investigation is a neuronal cell line known for its high lactate production compare to other cell lines. We, as well as others, have used this cell line and is considered an appropriate model to evaluate potential anti-cancer agents [22 (link), 23 (link)]. We also used the N2-A cell line to investigate the “Warburg Effect” phenomenon [24 (link)], and cancer cells metabolism [25 (link), 26 (link)]. On the other hand, the DI-TNC1 is an astrocyte immortal cell line with lower lactate efflux production compared to N2-A cells, an observation in our lab. The DI-TNC1 is very important in controlling brain energy metabolism [27 (link), 28 (link)]. Cell culture Dulbecco's Modified Eagle Medium (DMEM), fetal bovine serum (FBS), penicillin/streptomycin, DPBS, and trypsin were all from Atlanta Biologicals (Atlanta, GA, USA). Cells were cultured in 75-cm TC flask at 37°C in humidified 5% CO2 incubator and were subcultured as needed with trypsin/EDTA. Growing media was supplemented with 10% FBS (v/v), 4 mM L-glutamine, and 1% penicillin /streptomycin.
Messeha S.S., Zarmouh N.O., Taka E., Gendy S.G., Shokry G.R., Kolta M.G, & Soliman K.F. (2016). The Role of Monocarboxylate Transporters and Their Chaperone CD147 in Lactate Efflux Inhibition and the Anticancer Effects of Terminalia chebula in Neuroblastoma Cell Line N2-A. European journal of medicinal plants, 12(4), EJMP.23992.
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