Hela s3

Manufactured by Thermo Fisher Scientific

Sourced in United States

The HeLa S3 is a cell line derived from human cervical cancer cells. It is a widely used model system in cell biology research. The HeLa S3 cell line maintains a consistent growth rate and morphology, making it a reliable tool for various laboratory applications.

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HeLa S3 cells (2 citations)

9 protocols using hela s3

Fluorescent DNA Probe Labeling

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DNA and RNA strands were purchased from Integrated DNA Technology (IDT, Coralville, IA). Amine-modified DNA probes were labeled with Alexa-488, Cy3, or Cy5 mono NHS-esters by incubating 0.05 mM DNA with 3.0 mM fluorophores in a reaction buffer (100 mM Na₂BO₇ pH 8.5) for 3 h. The excess dyes were removed by using ethanol precipitation. Labeled oligonucleotides were stored in distilled water. Total RNA from human HeLa-S3 was purchased from Thermo Fisher Scientific (AM7852).

Kim J., Kang C., Shin S, & Hohng S. (2022). Rapid quantification of miRNAs using dynamic FRET-FISH. Communications Biology, 5, 1072.

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Cell Culture and IH Compound Preparation

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The human CC cell lines Hela S3 and SiHa were purchased from Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China), and the cells were maintained in RPMI 1640 (Thermo Fisher Scientific, Waltham, MA, USA) or DMEM (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific) and 100 mg/mL penicillin/streptomycin (Solarbio, Beijing, China) at 37°C in a humidified atmosphere with 5% CO₂. IH was kindly provided by Zhejiang Beta Pharma Co., Ltd. and dissolved in 100% dimethyl sulfoxide (DMSO) (Solarbio) to a final concentration of 50 μM and stored at −20°C.

Wang X., Gu Y., Liu H., Shi L, & Sun X. (2018). Icotinib hydrochloride enhances chemo- and radiosensitivity by inhibiting EGFR signaling and attenuating RAD51 expression and function in Hela S3 cells. OncoTargets and therapy, 11, 1245-1258.

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Sourcing Diverse Human RNA Samples

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Universal Human Reference RNA (UHRR) was purchased from Agilent, and HeLa S3 and MCF-7 RNAs were purchased from Thermo Fisher. RNAs from matched frozen healthy/tumor tissues of breast cancer patients were purchased from Origene (500 ng; Patient A: PR + , ER + , HER2 -, CR562524/CR543839; Patient B: PR unknown, ER -, HER2 -, CR560540/CR532030).

Yao J., Xu H., Ferrick-Kiddie E.A., Nottingham R.M., Wu D.C., Ares M., & Lambowitz A.M. (2020). Human cells contain myriad excised linear intron RNAs with links to gene regulation and potential utility as biomarkers.

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Culturing HeLa S3 and HEK293 T-REx Cells

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HeLa S3 and HEK293 T-REx (Thermo Fisher Scientific) cells were maintained in DMEM supplemented with 10% FBS in a humidified incubator at 5% CO2 and 37°C. The stable cell lines of HEK293 T-REx were selected following the manufacturer's instructions.

Chhipi-Shrestha J.K., Schneider‐Poetsch T., Suzuki T., Mito M., Khan K.M., Dohmae N., Iwasaki S., & Yoshida M. (2020). Splicing modulators elicit global translational repression by condensate-prone proteins translated from introns.

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Cell Line Cultivation and Maintenance

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H22 cells were obtained from the China Center for Type Culture Collection (ID: 3111C0001CCC000309, Wuhan, China). HEK293 (Cat# CRL-1573), Hela-S3 (Cat# CCL-2.2), 4T1 (Cat# CRL-2539), B16/F10 (Cat# CRL-6475), and CT26 (Cat# CRL-2638) cell lines were purchased from the American Type Culture Collection (ATCC; Manassas, USA). MC38 cells (RRID: CVCL_B288) were obtained from the National Cancer Institute (USA). HEK293, Hela-S3, 4T1, MC38, CT26, and B16/F10 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Cat# 11965092, Gibco-Thermo Fisher Scientific, USA) supplemented with 10% fetal bovine serum (FBS; Cat#16000044, Gibco). H22 cells were maintained in Roswell Park Memorial Institute 1640 medium (Cat# 11875093, Gibco) supplemented with 10% FBS. Under suspension conditions, Hela-S3 cells were maintained in a spinner flask (Jetbiofil, Guangzhou, China) in a serum-free medium (Cat# H740KJ, Basalmedia, Shanghai, China). All cells were cultured in an incubator at 37°C and 5% CO₂ atmosphere.

Zuo S., Wei M., Xu T., Kong L., He B., Wang S., Wang S., Wu J., Dong J, & Wei J. (2021). An engineered oncolytic vaccinia virus encoding a single-chain variable fragment against TIGIT induces effective antitumor immunity and synergizes with PD-1 or LAG-3 blockade. Journal for Immunotherapy of Cancer, 9(12), e002843.

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Cell Line Cultivation and Maintenance Protocol

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The American Type Culture Collection (ATCC; Manassas, USA) provided the cell lines HEK293 (CRL-1573), Hela-S3 (CCL-2.2), 4T1 (CRL-2539), and CT26 (CRL-2638). Panc02 and KPC cell lines were stored in our lab. 4T1^Hyal1 was constructed by stably transfecting 4T1 cells with a lentivirus carrying the Hyal1 gene. Dulbecco’s modified Eagle’s medium (DMEM; 11965092, ThermoFisher) supplemented with 10% fetal bovine serum (FBS; 16000044, Gibco) was used to sustain HEK293, Hela-S3, Panc02, KPC, 4T1, and CT26 cells. Hela-S3 cells were kept in suspension in a spinner flask in serum-free medium (H740KJ, Basalmedia, Shanghai, China). All cells were grown in an incubator set at 37°C and 5% CO₂.

Wang S., Li Y., Xu C., Dong J, & Wei J. (2024). An oncolytic vaccinia virus encoding hyaluronidase reshapes the extracellular matrix to enhance cancer chemotherapy and immunotherapy. Journal for Immunotherapy of Cancer, 12(3), e008431.

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Cell Line Culture Protocol

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The cell lines used in this study included HEK293 (human embryonic kidney cell), 4T1 (mouse mammary carcinoma cell), CT26 (mouse colon carcinoma cell), MC38 (mouse colon carcinoma cell), H22 (mouse hepatocellular carcinoma cell), and Hela-S3 (Human cervix carcinoma cell). HEK293 (Cat# CRL-1573, RRID: CVCL_0045), 4T1 (Cat# CRL-2539, RRID: CVCL_0125), CT26 (Cat# CRL-2638, RRID: CVCL_7256), and Hela-S3 (Cat# CCL-2.2, RRID: CVCL_0058) cells were obtained from the American Type Culture Collection (ATCC; USA). MC38 cells (RRID: CVCL_B288) were obtained from the National Cancer Institute (NCI; USA). H22 cells (ID: 3111C0001CCC000309) were obtained from the China Center for Type Culture Collection (CCTCC; Wuhan, China; http://cellresource.cn). HEK293, 4T1, CT26, MC38, and Hela-S3 cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Cat# 11965092, Gibco-Thermo Fisher Scientific, USA) supplemented with 10% fetal bovine serum (FBS; Cat#16000044, Gibco). H22 cells were cultured in RPMI 1640 medium (Cat# 11875093, Gibco) supplemented with 10% FBS (Cat#16000044, Gibco). Hela-S3 cells were cultured in suspension in serum-free medium (Cat# H740KJ, Basalmedia, Shanghai, China) in spinner flasks (Cat# TCB002002, Jetbiofil, Guangzhou, China). All cells were incubated at 37 °C with 5% CO₂ atmosphere.

Zuo S., Wei M., He B., Chen A., Wang S., Kong L., Zhang Y., Meng G., Xu T., Wu J., Yang F., Zhang H., Wang S., Guo C., Wu J., Dong J, & Wei J. (2021). Enhanced antitumor efficacy of a novel oncolytic vaccinia virus encoding a fully monoclonal antibody against T-cell immunoglobulin and ITIM domain (TIGIT). EBioMedicine, 64, 103240.

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Cell Line Maintenance and Acquisition

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The HEK293T and HeLaS3 cells were obtained from the American Type Culture Collection. The BTR cells were provided by Dr Daniel S. Peeper (Netherlands Cancer Institute). The p53+/+ and p53−/− HCT116 cells were provided by Lin Zhang. The U2OS cells were provided by Robert W. Sobol. The MCF10A and MCF7 cells were provided by Dr Yi Huang. The viral packaging line Pheonix-A was provided by Dr E.V. Prochownik. The HEK293T, HeLaS3, U2OS, MDA-MB-231 and Pheonix-A cells were maintained in DMEM (Invitrogen) with 10% FBS. The BTR cells were maintained as described previously11 (link). The HCT116 cells were maintained in McCoy's 5A (Invitrogen) with 10% FBS.

Hu D., Gur M., Zhou Z., Gamper A., Hung M.C., Fujita N., Lan L., Bahar I, & Wan Y. (2015). Interplay between arginine methylation and ubiquitylation regulates KLF4-mediated genome stability and carcinogenesis. Nature Communications, 6, 8419.

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Culturing LNCaP and HeLaS3 Cell Lines

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LNCaP (male, prostate carcinoma) and HeLaS3 (female, cervical adenocarcinoma) cell lines were obtained from the American Type Culture Collection. LNCaP cells were cultured in RPMI1640 medium, and HeLaS3 cells were cultured in Ham’s F-12K medium, both supplemented with 10% fetal bovine serum (FBS; Invitrogen) and 1% penicillin/streptomycin (Invitrogen). Cell lines were maintained at 37°C in a 5% CO₂ cell culture incubator. Cell lines were genotyped to confirm their identity at the University of Michigan Sequencing Core and tested routinely for Mycoplasma contamination.

Yi-Mi W., Cieslik M., Lonigro R.J., Pankaj V., Reimers M.A., Xuhong C., Yu N., Lisha W., Kunju L.P., de Sarkar N., Heath E.I., Jonathan C., Feng F.Y., Nelson P.S., de Bono J.S., Weiping Z., Bruce M., Alva A., Robinson D.R, & Chinnaiyan A.M. (2018). Inactivation of CDK12 delineates a distinct immunogenic class of advanced prostate cancer. Cell, 173(7), 1770-1782.e14.

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