SVG-A cells, HEK293A (CRL-1573), HEK293T (ATCC CRL-11268), Lenti-X HEK293T cells (takara, cat# 632180) and African green monkey (CV-1) (ATCC, CCL-70) cells were used. SVG-A cells are a third-generation sub clone of the original SVG cell line (Major et al., 1985 (link)) and express the astrocyte marker GFAP. SVG-A cells and CV-1 were cultured in MEM supplemented with 10% fetal bovine serum and 1% of Amphotericin B, Penicillin, Streptomycin solution. HEK293A and HEK293T were cultured in DMEM supplemented with 10% fetal bovine serum and 1% of Amphotericin B, Penicillin, Streptomycin solution. All cells were grown in an incubator at 37°c with 5% CO2. SVG-A cells and HEK293A cells were authenticated using the ATCC cell line authentication service (STR profile). They have a 55% match with SVG p.12 (human fetal glial cells, CRL-8621) from ATCC and they are of male origin. HEK293A cells have an 81% match with the parental line and are of female origin. We are in the process of authenticating the other cell lines used in the study.
Hek293a
Manufactured by American Type Culture Collection
Sourced in United States
The HEK293A is a cell line derived from human embryonic kidney cells. It is a widely used host cell line for the production and study of recombinant proteins, viral vectors, and other biological products.
Automatically generated - may contain errors
Lab products found in correlation
Hek293t, by American Type Culture Collection (22 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (9 mentions)
Hct116, by American Type Culture Collection (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions)
Hek293t, by Thermo Fisher Scientific (4 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (3 mentions)
Medium 199, by Merck Group (3 mentions)
Cos 7, by American Type Culture Collection (3 mentions)
H1299, by American Type Culture Collection (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Dmem f12, by Thermo Fisher Scientific (2 mentions)
Dexamethasone (dex), by Merck Group (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Lenti x hek293t cells, by Takara Bio (2 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (2 mentions)
Tnt coupled transcription translation system, by Promega (2 mentions)
Hepa1 6, by American Type Culture Collection (2 mentions)
53 protocols using hek293a
1
Authentication of Cell Lines for Research
2
Authentication of Cell Lines for Research
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SVG-A cells, HEK293A (CRL-1573), HEK293T (ATCC CRL-11268), Lenti-X HEK293T cells (takara, cat# 632180) and African green monkey (CV-1) (ATCC, CCL-70) cells were used. SVG-A cells are a third-generation sub clone of the original SVG cell line (Major et al., 1985 (link)) and express the astrocyte marker GFAP. SVG-A cells and CV-1 were cultured in MEM supplemented with 10% fetal bovine serum and 1% of Amphotericin B, Penicillin, Streptomycin solution. HEK293A and HEK293T were cultured in DMEM supplemented with 10% fetal bovine serum and 1% of Amphotericin B, Penicillin, Streptomycin solution. All cells were grown in an incubator at 37°c with 5% CO2. SVG-A cells and HEK293A cells were authenticated using the ATCC cell line authentication service (STR profile). They have a 55% match with SVG p.12 (human fetal glial cells, CRL-8621) from ATCC and they are of male origin. HEK293A cells have an 81% match with the parental line and are of female origin. We are in the process of authenticating the other cell lines used in the study.
3
Cell Line Propagation and Validation
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HEK293A, HeLa, and OVCAR3 cells were purchased from the American Type Culture Collection (ATCC); HEK293A and HeLa cells were cultured in DMEM with 10% fetal bovine serum, while OVCAR3 cells were cultured in RPMI with 10% fetal bovine serum. RPE1-hTERT FLAG-Cas9 TP53−/− BRCA1−/− cells were a gift that was kindly provided by Dr. Daniel Durocher (University of Toronto) and were cultured in DMEM with 10% fetal bovine serum. RMUGS was obtained from the Japanese Collection of Research Bioresources Cell Bank and cultured in Ham’s F12 medium with 10% fetal bovine serum. Knockout cells generated in 293 A and HeLa cells were created with pLentiCRISPRv2 (Addgene, #52961) containing indicated gRNAs (Figure 7—figure supplement 1 ), as described previously (Wang et al., 2021 (link)). All knockout cells were validated by western blotting and DNA sequencing (Figure 7—figure supplement 1 ). The 293A-derived PARP1/2 DKO cells were the same as those in the previous study (Wang et al., 2021 (link)). All cell lines were free of mycoplasma contamination.
4
Cell Line Cultivation and Validation Protocol
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HEK293A, HeLa, and OVCAR3 cells were purchased from the American Type Culture Collection (ATCC); HEK293A and HeLa cells were cultured in DMEM with 10% fetal bovine serum, while OVCAR3 cells were cultured in RPMI with 10% fetal bovine serum. RPE1-hTERT FLAG-Cas9 TP53−/− BRCA1−/− cells were a gift that was kindly provided by Dr. Daniel Durocher (University of Toronto) and were cultured in DMEM with 10% fetal bovine serum. RMUGS was obtained from the Japanese Collection of Research Bioresources Cell Bank and cultured in Ham’s F12 medium with 10% fetal bovine serum. Knockout cells generated in 293A and HeLa cells were created with pLentiCRISPRv2 (Addgene, #52961) containing indicated gRNAs (Fig. S8 ), as described previously33 (link). All knockout cells were validated by Western blotting and DNA sequencing (Fig. S8 ). The 293A-derived PARP1/2 DKO cells were the same as those in the previous study33 (link). All cell lines were free of mycoplasma contamination.
5
Cell Culture and Maintenance Protocol
6
Cell Culture and Transfection Protocols
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HEK293A (ATCC), HEK293T (ATCC), and HeLa (ATCC) cells were cultured in DMEM medium with 10% FBS. All the cells were cultured at 37 °C in cell culture incubator with humidified environment in 5% CO2. Lipofectamine 3000 (Invitrogen) or polythylenimine (PEI) (Polysciences) transfection reagents were used for plasmid transfection. All cell lines used were tested for mycoplasma contamination using the Mycoplasma Test Kit (Shanghai Yise Medical Technology Co. Ltd, PM008).
7
Cell Culture and Transfection of OCRL1
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Normal human proximal tubule epithelial (HK2) and human embryonic kidney epithelial 293T (HEK293T) and 293A (HEK293A) cells were purchased from ATCC. OCRL1 Knock-out (KO) HK2 and HEK293T cells were described earlier [25 (link)]; KO cell lines were previously validated [25 (link),30 (link)]. All cells were cultured in DMEM with streptomycin/penicillin, 2 mM L-glutamine and 10% fetal bovine serum (FBS) and maintained in an incubator at 37 °C with 5% CO2. For the expression of OCRL1, wild-type and mutant constructs, cells were transfected with FuGENE reagent (FuGENE 6 at a 6:1 reagent:DNA ratio or FuGENE 4K at a 4:1 reagent:DNA ratio, based on reagent availability (Promega)), following manufacturer protocols. In short, FuGENE was added to 100 μL of serum-free DMEM media, briefly vortexed, and incubated at room temperature for 5 min. Then, 1 μg of DNA was added to the reaction tube, briefly vortexed, and incubated at room temperature for 30 min. The final complex solution was added dropwise to the cells, and the plate was swirled to evenly spread the solution within the cell-containing wells.
8
Culturing Human Cell Lines for Research
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Human embryonic kidney 293T and 293A cells (HEK-293T and HEK-293A, respectively; ATCC, Manassas, VA, USA) and human cervical adenocarcinoma cells expressing a tetracycline-inducible repressor (HeLa Tet-Off; Clontech, Mountain View, CA, USA) were cultured in Dulbecco’s modified Eagle medium (DMEM; Gibco, Carlsbad, CA, USA) supplemented with 10% heat-inactivated fetal bovine serum (Gibco), 100 U/ml penicillin, 100 μg/ml streptomycin, and 2 mM l -glutamine (Gibco). Pooled human umbilical vein endothelial cells (HUVECs; Lonza, Basel, Switzerland) were cultured in endothelial cell growth medium 2 (EGM-2; Lonza). For HUVEC passaging, tissue culture plates were precoated for 30 min at 37°C with 0.1% (wt/vol) porcine gelatin (Sigma, St. Louis, Missouri, USA) in 1× phosphate-buffered saline (PBS; Gibco). All cells were routinely tested for mycoplasma by PCR method.
9
Cell Culture Protocol for HeLa and HEK293A
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The human cervical carcinoma cell line HeLa and human embryonic kidney cell line HEK293A were obtained from ATCC (Manassas, VA, USA). LATS1/2-DKO HEK293A cells were generously provided by Dr Kun-Liang Guan, the Department of Pharmacology and Moores Cancer Center at University of California San Diego (43 (link)). The cells were maintained in Dulbecco's modified Eagle medium (DMEM; WELGENE, Gyeongsan, Korea) supplemented with 10% fetal bovine serum (FBS; Gibco, Gaithersburg, MD, USA), 100 U/ml penicillin, and 100 μg/ml streptomycin (WELGENE). The cells were incubated at 37°C in a humidified atmosphere containing 5% CO2.
10
Characterization of Cell Lines
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U2 osteosarcoma [U2OS, American Type Culture Collection (ATCC), HTB-96], Hepa 1-6 (ATCC, CRL-1830), human embryonic kidney 293A (HEK293A), and MEF cell lines were used. U2OS (WT and AMPKα1/α2 DKO) cells were a gift from R. Shaw (Salk Institute for Biological Studies, San Diego, CA, USA). WT and JNK1/2 DKO MEFs were a gift from P. Angel (German Cancer Research Center, DKFZ, Heidelberg, Germany). All cell lines were maintained in DMEM (Gibco, #11995) supplemented with 10% heat-inactivated FBS and PenStrep (50 U/ml; Life Technologies, #15140122). HEK293A cells (Thermo Fisher Scientific, #R70507) were used to generate AVs. All the cell lines were maintained at humidified 37°C incubator containing 5% CO2.
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