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U2os osteosarcoma cells

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The U2OS osteosarcoma cells are a well-characterized, immortalized human cell line derived from the bone tissue of a 15-year-old female. These cells display an osteoblastic phenotype and are commonly used in research to study bone-related processes and diseases.

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Lab products found in correlation

Streptomycin, by Thermo Fisher Scientific (3 mentions) Penicillin, by Thermo Fisher Scientific (3 mentions) Bjhtert, by American Type Culture Collection (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Viafect, by Promega (1 mentions) Acetylated α tubulin antibody, by Merck Group (1 mentions) Vectashield containing dapi, by Vector Laboratories (1 mentions) Sodium pyruvate, by Merck Group (1 mentions) Penicillin streptomycin, by American Type Culture Collection (1 mentions) L glutamine, by Merck Group (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) Mccoy s 5a modified medium, by American Type Culture Collection (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Pc 3 cells, by Thermo Fisher Scientific (1 mentions) Dmem f12 glutamax medium, by Thermo Fisher Scientific (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Insulin, by Merck Group (1 mentions) Human imr 90 fibroblasts, by American Type Culture Collection (1 mentions)

Close spelling variants of this product

U2OS cells (152 citations) U2OS human osteosarcoma cells (13 citations) U2OS osteosarcoma cells (HTB-96 (2 citations)

12 protocols using u2os osteosarcoma cells

1

SPECC1L-GFP mutant expression in U2OS cells

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SPECC1L-GFP expression constructs containing either wildtype or Thr397Pro, Gly1083Ser, Gln415Pro and ΔCHD mutations were transfected into U2OS osteosarcoma cells (ATCC, Manassas, Virginia, USA) using Viafect (Promega, Madison, Wisconsin, USA) or TransIT (Mirus Bio, Madison, Wisconsin, USA) according to manufacturer’s protocol. U2OS cells were grown in standard DMEM supplemented with 10% fetal bovine serum (FBS). Transfections and immunostaining were carried out on coverslips in 24-well plates. Cells were fixed in 4% paraformaldehyde (PFA) for 10 min, and blocked in phosphate buffered saline (PBS) with 1% goat serum and 0.1% Tween. Acetylated α-tubulin antibody (Sigma, St Louis, Missouri, USA) was used at 1:1000 dilution. After staining, coverslips were mounted in VectaShield containing DAPI (Vector Labs, Burlingame, California, USA).
Kruszka P., Li D., Harr M.H., Wilson N.R., Swarr D., McCormick E.M., Chiavacci R.M., Li M., Martinez A.F., Hart R.A., McDonald-McGinn D.M., Deardorff M.A., Falk M.J., Allanson J.E., Hudson C., Johnson J.P., Saadi I., Hakonarson H., Muenke M, & Zackai E.H. (2014). Mutations in SPECC1L, encoding sperm antigen with calponin homology and coiled-coil domains 1-like, are found in some cases of autosomal dominant Opitz G/BBB syndrome. Journal of medical genetics, 52(2), 104-110.
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2

Culturing Melanoma, Osteosarcoma, and Endothelial Cells

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A375, malignant melanoma cells and U2OS osteosarcoma cells were obtained from ATCC. A375 cells were cultured in DMEM (Gibco #11965-092) medium plus 10% FBS at 37°C with 5% CO2. HUVEC cells were cultured in F-12K media (ATTC 30-2004) supplemented with 0.1 mg/ml heparin, 1% endothelial cell growth supplement (Sigma E2759-15MG) and 10% FBS (ATCC #30-2020) at 37°C with 5% CO2.
Scull C.E., Zhang Y., Tower N., Rasmussen L., Padmalayam I., Hunter R., Zhai L., Bostwick R, & Schneider D.A. (2019). Discovery of novel inhibitors of ribosome biogenesis by innovative high throughput screening strategies. The Biochemical journal, 476(15), 2209-2219.
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3

Culturing MIN6 and U2OS Cell Lines

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MIN6 pancreatic β-cells were a generous gift from Peter Arvan (University of Michigan) and cultured in DMEM with 4.5g/L glucose, L-glutamine, and sodium pyruvate (Sigma-Aldrich). Media was supplemented with 10% fetal bovine serum, 1% penicillin streptomycin solution, and 140 μmol/L β-mercaptoethanol (Sigma-Aldrich). U2OS osteosarcoma cells were purchased from ATCC and maintained in McCoy’s 5a Medium Modified (ATCC) supplemented with 10% fetal bovine serum and 1% penicillin streptomycin solution.
McKimpson W.M., Chen Y., Irving J.A., Zheng M., Weinberger J., Lek Wen Tan W., Tiang Z., Jagger A.M., Chua SC J.r., Pessin J.E., Foo R.S., Lomas D.A, & Kitsis R.N. (2021). Conversion of the death inhibitor ARC to a killer activates pancreatic β-cell death in diabetes. Developmental cell, 56(6), 747-760.e6.
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4

Culturing Immortalized Human Cell Lines

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Human telomerase expressing BJ (BJ-hTERT) skin fibroblasts and U2OS osteosarcoma cells were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA). The HeLa VST was provided by Dr. Roderick O’Sullivan (University of Pittsburgh). Cells were cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 50 units/mL penicillin, and 50 units/mL streptomycin (Gibco) at 37 °C in humidified chambers with 5% CO2 and 20% O2. Except BJ-hTERT cells were cultured with 10% FBS from Hyclone and at 5% O2, which improves proliferation of this cell line.
Orenstein A., Berlyoung A.S., Rastede E.E., Pham H.H., Fouquerel E., Murphy C.T., Leibowitz B.J., Yu J., Srivastava T., Armitage B.A, & Opresko P.L. (2017). γPNA FRET Pair Miniprobes for Quantitative Fluorescent In Situ Hybridization to Telomeric DNA in Cells and Tissue. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 22(12), 2117.
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5

Culturing Immortalized Human Cell Lines

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Human telomerase expressing BJ (BJ-hTERT) skin fibroblasts and U2OS osteosarcoma cells were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA). The HeLa VST was provided by Dr. Roderick O’Sullivan (University of Pittsburgh). Cells were cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 50 units/mL penicillin, and 50 units/mL streptomycin (Gibco) at 37 °C in humidified chambers with 5% CO2 and 20% O2. Except BJ-hTERT cells were cultured with 10% FBS from Hyclone and at 5% O2, which improves proliferation of this cell line.
Orenstein A., Berlyoung A.S., Rastede E.E., Pham H.H., Fouquerel E., Murphy C.T., Leibowitz B.J., Yu J., Srivastava T., Armitage B.A, & Opresko P.L. (2017). γPNA FRET Pair Miniprobes for Quantitative Fluorescent In Situ Hybridization to Telomeric DNA in Cells and Tissue. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 22(12), 2117.
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6

Prostate and Osteosarcoma Cell Culture

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PC-3 prostate bone metastasis and U2OS osteosarcoma cells were obtained from the American Type Culture Collection (ATCC, Manassas, Virginia, USA). PC-3 cells were propagated in RPMI-1640 medium (11875093, Gibco), U2OS cells were propagated in high glucose DMEM (10313021, Gibco), both supplemented with 10% fetal bovine serum (102701106, Gibco) and 1% penicillin–streptomycin (10378016, Gibco). All cultures were incubated at humidified 37 °C in 5% CO2.
Guerra Liberal F.D., Thompson S.J., Prise K.M, & McMahon S.J. (2023). High-LET radiation induces large amounts of rapidly-repaired sublethal damage. Scientific Reports, 13, 11198.
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7

CALUX Receptor Activation Assay

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CALUX cells based on human U2OS osteosarcoma cells (American Type Culture Collection [ATCC], Manassas, USA), expressed a full length PPARα,δ or γ receptor construct, respectively, and a luciferase reporter gene. 17 In addition, to assess non-specific luciferase modulation, e.g. due to cytotoxicity, the Cytotox CALUX cell line was used, consisting of U2-OS cells constitutively expressing the luciferase gene. 23 CALUX cells were cultured in a DMEM/F12 glutamax medium supplemented with 7.5% fetal calf serum, non-essential amino acids, and penicillin/streptomycin (all from Invitrogen, Breda, The Netherlands) at the final concentrations of 10 U mL -1 and 10 μg mL -1 , respectively, as described before. 24 Once per week, G418 (Duchefa Biochemie, Haarlem, The Netherlands) was added to the culture medium at a nominal concentration of 200 μg mL -1 .
Pham Ngoc L., Helmus R., Ehlers A.W., Swart K., Besselink H., de Rijke E., Dang Thi Cam H., Brouwer A, & van der Burg B. (2021). Effect-directed analysis and chemical identification of agonists of peroxisome proliferator-activated receptors in white button mushroom. Food & function, 12(1).
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8

Cell Culture Conditions for Various Cell Lines

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HMECs (Lonza cat # CC-2551) were immortalized with
hTERT, as reported (Solimini et al., 2012 (link)), and were maintained in MEGM (Lonza cat #
CC-3150). hTERT immortalized HPNEs (also known as HPDEs)
were the generous gift of Paul Campbell, and were maintained in a mixture of
1 part M3:BaseF medium (INCELL # M300F-500) with 3 parts DMEM supplemented
with 10% FBS, 100 units/mL penicillin, and 0.1 mg/mL streptomycin. U2OS
osteosarcoma cells (obtained directly from American Type Culture Collection,
HTB-96) were maintained in McCoy’s 5A medium supplemented with 10%
(v/v) FBS (Hyclone), 100 units/mL of penicillin, and 0.1 mg/mL streptomycin.
293T cells were maintained in DMEM supplemented with 10% (v/v) FBS
(Hyclone), 100 units/mL penicillin, and 0.1 mg/mL streptomycin. Human IMR90
fibroblasts were obtained from American Type Culture Collection and were
immortalized with hTERT. Cells were maintained in
DMEM-glutamax (Invigtrogen) supplemented with 15% FBS,
penicillin–streptomycin, and 0.1 mM nonessential amino acids
(Invitrogen). MCF7 cells were the generous gift of the Polyak laboratory,
and were maintained in DMEM supplemented with 10% FBS, 10 ug/mL Insulin
(Sigma #I0516-5mL), 100 units/mL penicillin, and 0.1 mg/mL streptomycin.
Sack L.M., Davoli T., Li M.Z., Li Y., Xu Q., Naxerova K., Wooten E.C., Bernardi R.J., Martin T.D., Chen T., Leng Y., Liang A.C., Scorsone K.A., Westbrook T.F., Wong K.K, & Elledge S.J. (2018). Profound Tissue Specificity in Proliferation Control Underlies Cancer Drivers and Aneuploidy Patterns. Cell, 173(2), 499-514.e23.
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9

Cell Culture Protocols for Osteosarcoma and Keratinocytes

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U2OS osteosarcoma cells (American Type Culture Collection, Manassas, VA) and HaCaT (immortalized skin keratinocytes) were grown in Dulbecco’s minimal essential medium with GlutaMAX (Invitrogen, Carlsbad, CA) and 10 % fetal bovine serum. OKF6-TERT2 cells (TERT-immortalized primary oral keratinocytes) were cultured in keratinocyte serum-free media (Invitrogen) supplemented with growth factors as per the manufacturer’s recommendations. The media were supplemented with 100 IU/mL penicillin and 100 IU/mL streptomycin (Invitrogen). Quality control was sporadically performed using the Mycoplasma Plus™ PCR Primer Set (Agilent Technologies, Santa Clara, CA).
Lin P., Mobasher M.E., Hakakian Y., Kakarla V., Naseem A.F., Ziai H, & Alawi F. (2015). Differential requirements for H/ACA ribonucleoprotein components in cell proliferation and response to DNA damage. Histochemistry and cell biology, 144(6), 543-558.
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10

Cell Culture Conditions for Cancer Cell Lines

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SW480 CRCs, A549 lung cancer cells, HeLa CCL2 cervical cancer cells, and U-2 OS osteosarcoma cells were obtained from the American Type Culture Collection (Manassas, VA). CCD-18Co cells were obtained from the Korean Cell Line Bank (Seoul, Korea).The SW480, CCD-18Co and HeLa CCL2 cells were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum (FBS), 100 μg/ml penicillin, and 100 μg/ml streptomycin. A549 cells were cultured in RPMI 1640 medium supplemented with 10% FBS, 100 μg/ml penicillin, and 100 μg/ml streptomycin. U-2 OS cells were cultured in McCoy's 5A medium supplemented with 10% FBS, 100 μg/ml penicillin, and 100 μg/ml streptomycin. The cultured cells were incubated under a humidified atmosphere of 5% CO2 at 37 °C.
Yu J.E., Yeo I.J., Lee D.W., Chang J.Y., Son D.J., Yun J., Han S.B, & Hong J.T. (2022). Snake venom induces an autophagic cell death via activation of the JNK pathway in colorectal cancer cells. Journal of Cancer, 13(12), 3333-3341.
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