The GBM cell line T98G and the LGG cell line Hs683 were obtained from ATCC. The cells were cultured in high glucose DMEM medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.) at 37°C under an atmosphere of 5% CO2 and 95% air. In addition, high glucose DMEM supplemented with 2% FBS was used in the scratch testing. The medium (half of the total volume) was changed every 3 days during the culture period.
Hs683
Manufactured by American Type Culture Collection
Sourced in United States
The HS683 is a laboratory equipment item produced by American Type Culture Collection. It is a device used for culturing and maintaining cell lines. The core function of the HS683 is to provide a controlled and sterile environment for cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
U87mg, by American Type Culture Collection (11 mentions)
Ln229, by American Type Culture Collection (10 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
U138mg, by American Type Culture Collection (4 mentions)
U118mg, by American Type Culture Collection (4 mentions)
Sw1783, by American Type Culture Collection (4 mentions)
Shg 44, by American Type Culture Collection (4 mentions)
Hek293t, by American Type Culture Collection (4 mentions)
Dulbecco modified eagle medium (dmem), by Corning (3 mentions)
Dbtrg 05mg, by American Type Culture Collection (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Sh sy5y, by American Type Culture Collection (2 mentions)
Antibiotic antimycotic solution, by Corning (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Sw1088, by American Type Culture Collection (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Hek293t cell, by American Type Culture Collection (2 mentions)
Q5 site directed mutagenesis, by New England Biolabs (1 mentions)
37 protocols using hs683
1
Glioma Cell Line Propagation Protocol
2
Glioblastoma Cell Culture Protocol
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Glioblastoma cells (A172, HS683, U87, U373, and T98G) were purchased from ATCC (Manassas, VA). Glioblastoma cells were grown in Dulbecco’s modified eagle medium (DMEM) containing 10% fetal bovine serum (FBS), 100 units/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified atmosphere containing 5% CO2. To collect glioblastoma-conditioned media (C.M.), glioblastoma cells were washed twice with PBS and cultured in serum-free DMEM for 36 h. Glioblastoma conditioned media were collected and centrifuged at 2000 r.p.m. for 10 min to remove cell debris.
3
Tau Plasmid Generation and Cloning
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Human tau plasmids have been previously generated and described [35 (link)]. S396, S404, S409 and S422 (according to 2N4R tau) were mutated to aspartic acid (phosphomimetic tau, PM-tau) using Q5 site-directed mutagenesis (New England Biolabs, (NEB)). The open reading frames (ORFs) of interaction candidates from yeast two-hybrid screening were PCR amplified from human cell lines cDNA libraries isolated from SH-SY5Y, HEK293T, U87MG and HS683 cells (ATCC). The amplified transcripts were first cloned into Gateway pENTR-SD-D-TOPO entry vector (Life Technologies), and further transferred into a pcDNA3.2-myc destination vector through LR recombination (Life Technologies). Mammalian expression constructs for V5 tagged tau and PM-tau were generated by LR recombination reaction into pcDNA3.2/V5-DEST vector (Life Technologies). For conventional cloning, PCR primers included restriction sites compatible with the cloning site of target vectors. Amplified products were cut with specific restriction enzymes (NEB) and ligated in-frame into pEGFP-C1 or pDNA3.1 vectors. All cloning primers are listed in Supplementary Table S3 .
4
Cell Line Maintenance for Viral Infection
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The African green monkey kidney Vero cell line (ATCC, Manassas, VA, USA), the human bronchial epithelial cell line 16HBE (ATCC), the human embryonic kidney cell line 293T (ATCC), the human glioma cell line Hs683 (ATCC), the human neuroblastoma cell line SH-SY5Y ATCC (and) the KMB17 cell line (IMB, CAMS, Yunnan, China) were maintained in high-glucose Dulbecco’s modified Eagle’s medium (DMEM; Corning, Corning, NY, USA) supplemented with 10% fetal bovine serum (HyClone, Logan, UT, USA). The culture medium was changed to DMEM supplemented with 2% fetal bovine serum after viral infection.
5
Culturing Normal and Glioblastoma Cell Lines
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6
Culturing Astrocytes and Glioma Cells
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The normal astrocytes and the glioma cell lines (U-87, U-118, M059K and Hs 683) were obtained from the ATCC, USA. The cells were cultured in RPMI 1640 (Thermo Scientific) medium supplemented with ampicillin and streptomycin (100 U/mL each) and 10% FBS at 37 °C in a humidified incubator containing 5% CO2.
7
Glioblastoma Cell Line Cultivation
8
Murine and Human Glioblastoma Cell Lines
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The murine astrocyte cell line C8-D1A was commercially obtained from Fuheng (FH0837, FuHeng Cell Center, Shanghai, China). Human GM U373, HS683 and U138MG cell lines were commercially obtained from ATCC. SHG44, U251, T98G, TJ905 and A172 cells were commercially obtained from Bolida (Bolida, Xuzhou, Jiangsu, China). The cells were cultured in DMEM containing 10% heat-inactivated FBS (FB25015, Clark, Virginia, USA), 100 mg/L streptomycin and 100 U/mL penicillin at 37 °C in a 5% CO2 environment. All human GM cell lines have been authenticated using STR profiling within the last three years. In addition, all experiments were performed with mycoplasma-free cells.
9
Glioma Cell Line Transfection Protocol
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The LGG glioma cell lines (SHG-44 and HS683) were purchased from ATCC, and human glial cells (HEB) were obtained from the Cancer Center, Sun Yat-Sen University. All cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM; HyClone, United States) supplemented with 10% fetal bovine serum at 37°C in a 5% CO2 incubator.
Small interfering RNAs (siRNAs) against target genes were synthesized by GenPharma (Suzhou, China). Cells were transfected using the Lipofectamine® RNAiMAX Transfection Reagent (Invitrogen, Carlsbad, California, United States) according to the manufacturer’s instructions. The siRNA sequences are listed inTable S1
Small interfering RNAs (siRNAs) against target genes were synthesized by GenPharma (Suzhou, China). Cells were transfected using the Lipofectamine® RNAiMAX Transfection Reagent (Invitrogen, Carlsbad, California, United States) according to the manufacturer’s instructions. The siRNA sequences are listed in
10
Glioblastoma Cell Line Cultivation
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LN229 (CRL-2611), U87 MG (HTB-14), U138 MG (HTB-16), Hs683 (HTB-138), DBTRG-05MG (CRL-2020), LN18 (CRL-2610), T98G (CRL-1690), A172 (CRL-1620) and U118MG (HTB15) human glioblastoma cell lines were from ATCC and cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Corning, 10–013-CV) supplemented with 10% Fetal Bovine Serum (Gibco, 10437028) and 1% Antibiotic-Antimycotic Solution (Corning, 30–004-CI) at 37°C with 5% CO2. The cell lines were not independently authenticated in this study. The cell lines were examined to be mycoplasma negative before experiments. Unless otherwise indicated, experiments were performed with cells grown to 50% confluency.
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