Htb 148

Manufactured by American Type Culture Collection

Sourced in United States

The HTB-148 is a laboratory equipment product designed for cell culture applications. It serves as a specialized incubator for maintaining and cultivating cell lines under controlled environmental conditions.

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Lab products found in correlation

Ptet off plasmid, by Takara Bio (1 mentions) Hek293t, by American Type Culture Collection (1 mentions) High glucose dmem, by Thermo Fisher Scientific (1 mentions) Cholesterol, by Merck Group (1 mentions) Pmscv pig, by Addgene (1 mentions) Bafilomycin, by Cayman Chemical (1 mentions) 2 hydroxypropyl β cyclodextrin hpβcd, by Merck Group (1 mentions) N2 supplement, by Thermo Fisher Scientific (1 mentions) Primary human astrocytes, by ScienCell (1 mentions) Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions) Tetracycline, by Merck Group (1 mentions) Glial cell derived neurotrophic factor, by Thermo Fisher Scientific (1 mentions) Dibutyryl camp, by Thermo Fisher Scientific (1 mentions) One shot fetal bovine serum, by Thermo Fisher Scientific (1 mentions) Sodium arsenate, by Merck Group (1 mentions)

Spelling variants of this product

H4 ATCC ® HTB-148™ Homo sapiens brain neuroglioma cells (2 citations) ATCC®HTB-148™) (2 citations)

4 protocols using htb 148

Generation of Isogenic H4 Cell Lines

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Human H4 neuroglioma cells (HTB-148, ATCC, USA) were transfected with the pTet-Off plasmid (Clontech, USA) and a stable clone was selected by geneticin resistance. The resulting H4 Tet-Off cell line was subsequently transfected with the pFRT/LacZeo plasmid (Life tech) to generate H4 TetOff FRT cell lines. A stable single integrant clone with high beta-galactosidase activity and zeocin resistance was then selected. H4 TetOff FRT empty cells were cotransfected with the pOG44 plasmid (Life tech), for transient expression of the Flp recombinase, and with either pTRE3G-BI-FRT-Hygro-V1S&SV2, pTRE3G-BI-FRT-Hygro-SL1&SL2, pTRE3G-BI-FRT-Hygro-V1Lz&LzV2 or pTRE3G-BI-FRT-Hygro-LzL1&LzL2 for generation of the H4 V1S&SV2, H4 SL1&SL2, H4 V1Lz&LzV2 or H4 LzL1&LzL2 cell lines, respectively. Stable clones resistant to hygromycin B but not to zeocin were then selected, confirming the recombinase-conducted insertion of our genes of interest at the FRT site, thus guaranteeing the isogenicity of our cell lines.

Moussaud S., Malany S., Mehta A., Vasile S., Smith L.H, & McLean P.J. (2015). Targeting alpha-synuclein oligomers by protein-fragment complementation for drug discovery in synucleinopathies. Expert opinion on therapeutic targets, 19(5), 589-603.

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TFEB Regulation in α-Synuclein Model

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2-hydroxypropyl-β-cyclodextrin (HPβCD) and cholesterol were purchased from Sigma-Aldrich, bafilomycin was from Cayman Chemical, and DAPI nuclear stain was from Enzo Life Sciences. TFEB siRNA (Cat. No. SI00094969) and control siRNA (Cat. No. 1027280) were purchased from Qiagen. pMSCV-PIG, gag-pol, and VSVG plasmids were from Addgene and TFEB-3XFLAG plasmid was a generous gift from Dr. Marco Sardiello (Baylor College of Medicine, Houston, TX)
H4 cells stably transfected for the expression of α-syn-EmGFP (H4/α-syn-GFP) were generated as previously described [34 (link),35 (link)]. Human H4 neuroglioma (HTB-148, ATCC), H4/α-syn-GFP, and HEK-293T cells (CRL-11268, ATCC) were cultured in high glucose DMEM (Fisher Scientific) supplemented with 10% fetal bovine serum and 1% PSQ, and maintained at 37°C and 5% CO₂.

Kilpatrick K., Zeng Y., Hancock T, & Segatori L. (2015). Genetic and Chemical Activation of TFEB Mediates Clearance of Aggregated α-Synuclein. PLoS ONE, 10(3), e0120819.

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Differentiation of Dopaminergic and Astrocytic Cells

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Primary human astrocytes (#1800, ScienCell Research Laboratories) were cultured as previously described (Pajarillo, Digman, et al., 2021 (link)) with a slight modification. Lund human mesencephalic (LUHMES, CRL-2927, ATCC) dopaminergic-like cells were maintained in DMEM/F-12 with 1% N2 supplement (Gibco) and 40 ng/ml basic fibroblast growth factor (PeproTech) and subcultured on culture flasks precoated with 50 μg/ml poly-l-ornithine and 1 μg/ml fibronectin. After plated in 24-well or 6-well tissue culture plates, LUHMES cells were differentiated into morphologically and biochemically mature dopamine-like neurons with a differentiation media containing tetracycline (MilliporeSigma), glial cell–derived neurotrophic factor, and dibutyryl-cAMP (PeproTech), followed by treatment with designated compounds and downstream analysis. Human H4 astrocytes (HTB-148, ATCC) were maintained in DMEM supplemented with 10% fetal bovine serum, 100 U/mL of penicillin, and 100 μg/mL of streptomycin and maintained at 37°C in a 95% air, 5% CO₂ incubator. Mouse cath.a-differentiated (CAD) catecholaminergic-like cells (MilliporeSigma) were differentiated into morphologically and biochemically mature neurons by replacing them with serum-free media.

Pajarillo E., Demayo M., Digman A., Nyarko-Danquah I., Son D.S., Aschner M, & Lee E. (2022). Deletion of RE1-silencing transcription factor in striatal astrocytes exacerbates manganese-induced neurotoxicity in mice. Glia, 70(10), 1886-1901.

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PEG10 Localization in Stress Granules

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H4 cells (H4 (ATCC® HTB-148) or neurons were cultured in Dulbecco’s Modified Eagle’s Medium, Catalog No. 30-2002 media supplemented with 10% FBS (One Shot Fetal Bovine Serum, GIBCO). Cells were seeded in a 96 well plate (Falcon® 96-well Black/Clear Flat Bottom) at 1500 cells per well. On the next day, they were transduced using PEG10- RF1/2-GFP or PEG10- RF1-GFP or just GFP construct at an MOI of 2 overnight and cell culture media was replaced the next day prior to treatments. At day 3, cells were either treated with 5 mM sodium arsenate (Sigma) or PBS for 1 hr. Post treatment, cells were immediately fixed in 4% PFA for 15 min and stained for PEG10 (using anti-GFP antibody), anti-G3BP1 antibody and DAPI to label the nuclei as described.

Pandya N.J., Wang C., Costa V., Lopatta P., Meier S., Zampeta F.I., Punt A.M., Mientjes E., Grossen P., Distler T., Tzouros M., Martí Y., Banfai B., Patsch C., Rasmussen S., Hoener M., Berrera M., Kremer T., Dunkley T., Ebeling M., Distel B., Elgersma Y, & Jagasia R. (2021). Secreted retrovirus-like GAG-domain-containing protein PEG10 is regulated by UBE3A and is involved in Angelman syndrome pathophysiology. Cell Reports Medicine, 2(8), 100360.

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