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Mimcd3

Manufactured by American Type Culture Collection
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The MIMCD3 is a laboratory equipment product offered by the American Type Culture Collection. It is designed for cell culture and related applications. The core function of the MIMCD3 is to provide a controlled environment for the cultivation and maintenance of cells.

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Lab products found in correlation

Htert rpe1, by American Type Culture Collection (5 mentions) Fibroblast growth medium, by Genlantis (4 mentions) Sh sy5y, by American Type Culture Collection (4 mentions) Hek293t, by American Type Culture Collection (2 mentions) Penicillin and streptomycin, by Thermo Fisher Scientific (2 mentions) Hct116, by American Type Culture Collection (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Dmem f12, by Thermo Fisher Scientific (1 mentions) Glutamine, by PanEco (1 mentions) Fluorodish, by World Precision Instruments (1 mentions) Hyclone, by Thermo Fisher Scientific (1 mentions) Opti mem, by Thermo Fisher Scientific (1 mentions) Fugene hd, by Promega (1 mentions) Mdckii, by American Type Culture Collection (1 mentions) Rpe 1, by American Type Culture Collection (1 mentions) Rpe 1, by Corning (1 mentions) Dulbecco modified eagle medium (dmem), by Corning (1 mentions) Penicillin streptomycin, by Cytiva (1 mentions) Hek293t, by Corning (1 mentions) Dmem f12, by Cytiva (1 mentions)

18 protocols using mimcd3

1

Characterization of Diverse Cell Lines

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Mouse inner medullary collecting duct (mIMCD3), human telomerase reverse transcriptase-transformed retinal pigment epithelium (hTERT-RPE1) and human neuroblastoma (SH-SY5Y) cells were derived from American Type Culture Collection (ATCC). The genomic status of the cell-lines were assessed by array CGH and karyotyping, and SH-SY5Y cells were authenticated by STR profiling (May 2013). All cell-lines were tested every three months for mycoplasma. Cell-lines were maintained in DMEM/Ham’s F12 medium supplemented with 10% foetal calf serum (FCS), under standard conditions (37°C, 5% CO2). Cells were passaged at a split ratio of 1:10 twice a week, with low passages (<25) for both mIMCD3 and hTERT-RPE1 cells. mIMCD3 cells were obtained from ATCC at passage 13 and were used for screening purposes between passage 17 and 25. SH-SY5Y cells were differentiated over 6 days by retinoic acid (RA) treatment followed by brain derived neurotrophic factor (BDNF) treatment. Cells were plated in standard culture medium for 24 hours, in standard medium containing 10μM RA for 3 days and in serum-free medium containing 50ng/ml BDNF cells for 3 days. Human dermal fibroblasts were derived from skin biopsies and cultured in fibroblast growth medium (Genlantis). hTERT-RPE1 cells were serum starved in normal media with 0.2% FCS for 48 hours to induce ciliogenesis.
Wheway G., Schmidts M., Mans D.A., Szymanska K., Nguyen T.M., Racher H., Phelps I.G., Toedt G., Kennedy J., Wunderlich K.A., Sorusch N., Abdelhamed Z.A., Natarajan S., Herridge W., van Reeuwijk J., Horn N., Boldt K., Parry D.A., Letteboer S.J., Roosing S., Adams M., Bell S.M., Bond J., Higgins J., Morrison E.E., Tomlinson D.C., Slaats G.G., van Dam T.J., Huang L., Kessler K., Giessl A., Logan C.V., Boyle E.A., Shendure J., Anazi S., Aldahmesh M., Al Hazzaa S., Hegele R.A., Ober C., Frosk P., Mhanni A.A., Chodirker B.N., Chudley A.E., Lamont R., Bernier F.P., Beaulieu C.L., Gordon P., Pon R.T., Donahue C., Barkovich A.J., Wolf L., Toomes C., Thiel C.T., Boycott K.M., McKibbin M., Inglehearn C.F., Stewart F., Omran H., Huynen M.A., Sergouniotis P.I., Alkuraya F.S., Parboosingh J.S., Innes A.M., Willoughby C.E., Giles R.H., Webster A.R., Ueffing M., Blacque O., Gleeson J.G., Wolfrum U., Beales P.L., Gibson T., Doherty D., Mitchison H.M., Roepman R, & Johnson C.A. (2015). An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes. Nature cell biology, 17(8), 1074-1087.
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2

Characterization of Diverse Cell Lines

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Mouse inner medullary collecting duct (mIMCD3), human telomerase reverse transcriptase-transformed retinal pigment epithelium (hTERT-RPE1) and human neuroblastoma (SH-SY5Y) cells were derived from American Type Culture Collection (ATCC). The genomic status of the cell-lines were assessed by array CGH and karyotyping, and SH-SY5Y cells were authenticated by STR profiling (May 2013). All cell-lines were tested every three months for mycoplasma. Cell-lines were maintained in DMEM/Ham’s F12 medium supplemented with 10% foetal calf serum (FCS), under standard conditions (37°C, 5% CO2). Cells were passaged at a split ratio of 1:10 twice a week, with low passages (<25) for both mIMCD3 and hTERT-RPE1 cells. mIMCD3 cells were obtained from ATCC at passage 13 and were used for screening purposes between passage 17 and 25. SH-SY5Y cells were differentiated over 6 days by retinoic acid (RA) treatment followed by brain derived neurotrophic factor (BDNF) treatment. Cells were plated in standard culture medium for 24 hours, in standard medium containing 10μM RA for 3 days and in serum-free medium containing 50ng/ml BDNF cells for 3 days. Human dermal fibroblasts were derived from skin biopsies and cultured in fibroblast growth medium (Genlantis). hTERT-RPE1 cells were serum starved in normal media with 0.2% FCS for 48 hours to induce ciliogenesis.
Wheway G., Schmidts M., Mans D.A., Szymanska K., Nguyen T.M., Racher H., Phelps I.G., Toedt G., Kennedy J., Wunderlich K.A., Sorusch N., Abdelhamed Z.A., Natarajan S., Herridge W., van Reeuwijk J., Horn N., Boldt K., Parry D.A., Letteboer S.J., Roosing S., Adams M., Bell S.M., Bond J., Higgins J., Morrison E.E., Tomlinson D.C., Slaats G.G., van Dam T.J., Huang L., Kessler K., Giessl A., Logan C.V., Boyle E.A., Shendure J., Anazi S., Aldahmesh M., Al Hazzaa S., Hegele R.A., Ober C., Frosk P., Mhanni A.A., Chodirker B.N., Chudley A.E., Lamont R., Bernier F.P., Beaulieu C.L., Gordon P., Pon R.T., Donahue C., Barkovich A.J., Wolf L., Toomes C., Thiel C.T., Boycott K.M., McKibbin M., Inglehearn C.F., Stewart F., Omran H., Huynen M.A., Sergouniotis P.I., Alkuraya F.S., Parboosingh J.S., Innes A.M., Willoughby C.E., Giles R.H., Webster A.R., Ueffing M., Blacque O., Gleeson J.G., Wolfrum U., Beales P.L., Gibson T., Doherty D., Mitchison H.M., Roepman R, & Johnson C.A. (2015). An siRNA-based functional genomics screen for the identification of regulators of ciliogenesis and ciliopathy genes. Nature cell biology, 17(8), 1074-1087.
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3

Culturing Human Kidney Cells

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Human kidney epithelial cells (HEK 293T, ATCC CRL-3216) and mIMCD-3 (ATCC CRL-2123) were cultured in DMEM and DMEM-F12 (Life Technologies) supplemented with 10% FBS respectively. For serum starvation mIMCD-3 cells were cultured in DMEM-F12 without FBS supplementation. Transfections were performed with Lipofectamine 2000 using recommended manufacturer conditions.
Scott C.A., Marsden A.N., Rebagliati M.R., Zhang Q., Chamling X., Searby C.C., Baye L.M., Sheffield V.C, & Slusarski D.C. (2017). Nuclear/cytoplasmic transport defects in BBS6 underlie congenital heart disease through perturbation of a chromatin remodeling protein. PLoS Genetics, 13(7), e1006936.
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4

Establishing Tetracycline-Inducible SMARCB1 Cell Line and CRISPR-Cas9 Engineered Renal Tumor Model

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mIMCD-3 (mouse inner medulla collecting duct) mouse cell lines were purchased from ATCC.
RMC219-Tet-inducible SMARCB1 cell line: The tetracycline-inducible pIND20-fSNF5-HA vector (35 (link)) was kindly donated by Bernard E. Weissman. The pInducer20 empty backbone (36 (link)) was a gift from Stephen Elledge (Addgene plasmid # 44012). Lentivirus was generated in HEK-293T cells and used to generate stable tet-inducible cell lines as previously described (37 (link)). RMC219 (also called JHRCC219), a human tumor cell line that was established in a previous study (23 (link)), was infected with tetracycline-inducible pIND20-fSNF5-HA vector. Cells were treated with 2 μg DOX for 3 d to reconstitute SMARCB1 (SNF5).
MSRT1 (Melinda Soeung renal tumor 1) was generated using CRISPR-Cas9 gene editing technology as described in SI Appendix, Figs. S2 and S3 and Supporting methods.
Soeung M., Perelli L., Chen Z., Dondossola E., Ho I.L., Carbone F., Zhang L., Khan H., Le C.N., Zhu C., Peoples M.D., Feng N., Jiang S., Zacharias N.M., Minelli R., Shapiro D.D., Deem A.K., Gao S., Cheng E.H., Lucchetti D., Walker C.L., Carugo A., Giuliani V., Heffernan T.P., Viale A., Tannir N.M., Draetta G.F., Msaouel P, & Genovese G. (2023). SMARCB1 regulates the hypoxic stress response in sickle cell trait. Proceedings of the National Academy of Sciences of the United States of America, 120(21), e2209639120.
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5

Optimizing Transfection Conditions for Live-Cell Imaging

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HeLa Kyoto and HEK293T cell lines were obtained from established frozen stocks of laboratory. Rat cardiomyoblasts H9c2 (ATCC CRL-1446) and mouse inner medullary collecting duct mIMCD-3 (ATCC CRL-2123) cell lines were obtained from ATCC. Mouse myoblasts C2C12 were a kind gift of N. Podkiychenko (Institute of Experimental Cardiology, National Medical Research Center for Cardiology, Moscow, Russia). All cell lines were grown in an adhesion culture in Dulbecco’s Modified Essential Medium (DMEM) with 2 mM glutamine and 4.5 g/l glucose (PanEco, Russia) supplemented with 10% fetal bovine serum (HyClone, ThermoScientific, USA) and 1% Penicillin + Streptomycin in a humidified atmosphere under 37°C and 5% CO2.
Twenty-four hours before transfection, cells were seeded on glass bottom culture dishes (Fluorodish, World Precision Instruments, USA). For transient transfection, the transfection reagent FuGene HD (Promega, USA) was used in accordance with the the manufacturer’s protocol. The transfection mixture was diluted with 700 μl of OptiMEM (Gibco, ThermoScientific, USA), and incubation was continued for 16–24 h.
After pre-optimization (data not shown) for wide-field and confocal microscopy, the DNA ratio for localized KECs and cytosolic pool was 2 for better contrast, for super-resolution microscopy DNA ratio was 0.5.
Perfilov M.M., Gurskaya N.G., Serebrovskaya E.O., Melnikov P.A., Kharitonov S.L., Lewis T.R., Arshavsky V.Y., Baklaushev V.P., Mishin A.S, & Lukyanov K.A. (2020). Highly photostable fluorescent labeling of proteins in live cells using exchangeable coiled coils heterodimerization. Cellular and molecular life sciences : CMLS, 77(21), 4429-4440.
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6

Lentiviral Preparation and Cell Line Assays

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293T/17 (ATCC Cat# CRL-11268, RRID:CVCL_1926) cells were used for lentiviral preparation and the Luciferase and endogenous gene activation assays. mIMCD3 (ATCC Cat# CRL-2123, RRID:CVCL_0429) cells were used to test Ccna1 antibody specificity. Cells were purchased from ATCC and were assumed to be authenticated by the supplier. Cells were not specifically monitored for mycoplasma contamination, but routine DAPI staining would have revealed the presence of contamination.
Vladar E.K., Stratton M.B., Saal M.L., Salazar-De Simone G., Wang X., Wolgemuth D., Stearns T, & Axelrod J.D. (2018). Cyclin-dependent kinase control of motile ciliogenesis. eLife, 7, e36375.
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7

Cell Culture of Common Cell Lines

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MDCK II (ATCC), hTERT RPE-1 (ATCC), and mIMCD3 (ATCC) cells were cultured at 37°C with 5% CO2 in DMEM with 10% FBS and penicillin/streptomycin. Cell lines were authenticated through STR testing. Each line was checked against the ATCC and DSMZ databases for fingerprinting. Cells were routinely screened for mycoplasma.
Jewett C.E., Soh A.W., Lin C.H., Lu Q., Lencer E., Westlake C.J., Pearson C.G, & Prekeris R. (2021). RAB19 directs cortical remodeling and membrane growth for primary ciliogenesis. Developmental cell, 56(3), 325-340.e8.
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8

Culturing MDCK, mIMCD3, RPE1 and HEK293T cells

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The MDCK, mIMCD3, RPE1 and HEK293T cell lines were obtained from ATCC (Manassas, VA, USA); details of these cell lines are listed in Table S1. Cells were maintained at 37°C with 5% CO2 in complete medium. For MDCK, RPE1 and HEK293T cells, the complete medium consisted of DMEM (#10-017-CV, Corning, Corning, NY, USA) with 10% fetal bovine serum (FBS) (#PS-300, Phoenix Scientific, San Marcos, CA, USA) and 1× penicillin-streptomycin (#30-002-CL, Corning). For mIMCD3 cells, the complete medium consisted of DMEM/F12 (#SH30023.01, Cytiva, Marlborough, MA, USA) with 10% FBS and 1× penicillin-streptomycin.
Hoffman H.K, & Prekeris R. (2023). HOPS-dependent lysosomal fusion controls Rab19 availability for ciliogenesis in polarized epithelial cells. Journal of Cell Science, 137(5), jcs261047.
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9

Cultivation and Characterization of Murine Cell Lines

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The 8A7H5 hybridoma (rat IgG2b, κ) was previously generated by immunization of rats with MuPyV VP1 virus-like particles (Swimm et al., 2010 (link)). NMuMG, BALB/3T3 clone A31 ‘A31’, and mIMCD-3 cells were purchased from ATCC. Mouse embryonic fibroblasts (MEFs) were isolated from day 13 C57BL/6 embryos. Hybridomas 8A7H5 and H35-17.2 (anti-CD8β) (Pierres et al., 1982 (link)) were maintained in PFHM-II Protein-Free Hybridoma Medium (ThermoFisher) at 37°C in 5% CO2. mAb was generated by growing the hybridomas in CELLine disposable bioreactor flasks (Corning). All other cells were maintained in Dulbecco’s Minimal Eagle Media supplemented with 10% fetal bovine serum, 100 U/mL penicillin, and 100 U/mL streptomycin (DMEM) at 37°C in 5% CO2. The sex of NMuMG cells is female, the sex of mIMCD-3 and A31 cells is not reported. Cell lines were mycoplasma negative, authenticated by STR profiling (ATCC), examined for correct cell morphology, and used at low passage number.
Lauver M.D., Goetschius D.J., Netherby-Winslow C.S., Ayers K.N., Jin G., Haas D.G., Frost E.L., Cho S.H., Bator C.M., Bywaters S.M., Christensen N.D., Hafenstein S.L, & Lukacher A.E. (2020). Antibody escape by polyomavirus capsid mutation facilitates neurovirulence. eLife, 9, e61056.
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10

Isogenic cell lines for Tsc1/Tsc2 knockout

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We used murine kidney-derived principal cell lines, i.e., mIMCD3, purchased from ATCC (ATCC® CRL-2123TM, Manassas, VA, United States). We previously developed derivative isogenic cell lines by knocking out the Tsc1 or Tsc2 gene by CRISPR/Cas9, and the derivative lines were designated T1G and T2J, respectively (Bissler et al., 2019 (link)). The Tsc1 gene was knocked out by targeting exon 4 (Bissler et al., 2019 (link)). Cystic kidney-derived epithelial cells, i.e., M1 cells, were also purchased from ATCC® (CRL-2038TM, Manassas, VA, United States). mIMCD3, T1G, and T2J cells were maintained in DMEM/F12 with 10% FBS, whereas M1 cells were maintained with DMEM/F12 with 5% FBS and supplemented with 5 μM dexamethasone. Cultures were maintained at 37°C in a humidified 95% air and 5% CO2 atmosphere.
Kumar P., Zadjali F., Yao Y., Siroky B., Astrinidis A., Gross K.W, & Bissler J.J. (2021). Tsc Gene Locus Disruption and Differences in Renal Epithelial Extracellular Vesicles. Frontiers in Physiology, 12, 630933.
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