Kyou dxr0109b

Manufactured by American Type Culture Collection

Sourced in United States

The KYOU-DXR0109B is a compact and versatile laboratory instrument designed for Raman spectroscopy applications. It features a powerful diode laser source and a high-performance CCD detector, enabling efficient data collection and analysis. The core function of this product is to provide researchers with a reliable tool for molecular identification and structural analysis.

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9 protocols using kyou dxr0109b

Differentiation and Culture of iPSC-Derived Motor Neurons

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The control human induced pluripotent stem cells (iPSCs) used in this study were obtained from ATCC (#KYOU-DXR0109B). The patient-derived iPSCs and their isogenic controls were previously described^{2 (link),75 (link)}. All iPSCs were cultured on Geltrex LDEV-Free, hESC-Qualified basement membrane matrix, supplemented with 1X Essential 8 supplement. Colonies were regularly passaged using 0.5 mM EDTA (15575–020, Invitrogen) in Dulbecco’s phosphate-buffered saline (DPBS). The cultures were routinely monitored for mycoplasma contamination by PCR. Motor neurons were generated from the iPSC lines using the previously published protocol^{25 (link)}.

Provasek V.E., Kodavati M., Guo W., Wang H., Boldogh I., Van Den Bosch L., Britz G, & Hegde M. (2023). lncRNA Sequencing Reveals Neurodegeneration-associated FUS Mutations Alter Transcriptional Landscape of iPS Cells That Persists In Motor Neurons. Research Square.

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Derivation of Neural Progenitor Cells from Human iPSCs

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Human iPSCs (KYOU-DXR0109B (201B7)) were purchased from ATCC and were grown using cell matrix basement membrane gel (ACS-3035) and pluripotent stem cell SFM XF/FF cell culture media (ACS-3002) at 37 °C and 5% CO₂. NPSCs were derived from iPSCs using PSC neural induction medium according to the manufacturer’s instructions. NPCs were generated by replacing E8M with neural induction medium about 24 h after passing iPSCs, and then these cells were maintained for days. Derived NPSCs (P0) were then passaged onto six-well plates coated with Geltrex (Thermo Fisher), and the lineage was expanded in stemPRO neural stem cell SFM media (A1050901). Finally, the efficiency of neural induction was determined at passage 3 by IF staining with Nestin.^{52 (link),53 (link)} Neurons were generated from iPSC cells as described previously (Figure S1e).^{53 (link)}

Dharmalingam P., Talakatta G., Mitra J., Wang H., Derry P.J., Nilewski L.G., McHugh E.A., Fabian R.H., Mendoza K., Vasquez V., Hegde P.M., Kakadiaris E., Roy T., Boldogh I., Hegde V.L., Mitra S., Tour J.M., Kent T.A, & Hegde M.L. (2020). Pervasive Genomic Damage in Experimental Intracerebral Hemorrhage: Therapeutic Potential of a Mechanistic-Based Carbon Nanoparticle. ACS nano, 14(3), 2827-2846.

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Routine Culture of hu-iPSCs on Matrigel

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KYOU-DXR0109B (ACS-1023; ATCC) hu-iPSCs were routinely cultured on low growth factor Matrigel (Roche) in mTeSR1 medium (Stem Cell Technologies) with 5% CO₂ in a humidified incubator as described in the mTeSR handbook. Colonies were passaged at approximately 70–80% confluency before colonies had started to contact each other.

Groveman B.R., Foliaki S.T., Orru C.D., Zanusso G., Carroll J.A., Race B, & Haigh C.L. (2019). Sporadic Creutzfeldt-Jakob disease prion infection of human cerebral organoids. Acta Neuropathologica Communications, 7, 12.

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Reprogramming Adult Cells to Induced Pluripotent Stem Cells

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iPS-KYOU, has been obtained in the Shinya Yamanaka laboratory (Kyoto University, Japan) by retroviral reprogramming of adult female skin fibroblasts. The iPS-KYOU cell line was purchased from the ATCC cell bank (KYOU-DXR0109B, ACS-1023™, ATCC®, United States).
iPS-AFS17, human Induced Pluripotent Stem Cells, obtained by lentiviral reprogramming of amniotic fluid stem cells by Dashinimaev et al. (2017) (link).
iPS-DP human Induced Pluripotent Stem Cells, obtained by lentiviral reprogramming of dermal papilla cells (Muchkaeva et al., 2014 (link)).
iPS-DYP0730, human Induced Pluripotent Stem Cells, derived from dermal fibroblasts obtained from ATCC CCL-54 Detroit 532, a human Down syndrome cell line. The iPS-DYP0730 cell line was purchased from the ATCC cell bank (ACS-1003™, ATCC®, United States).
For all pluripotent stem cell passaging, we used ACCUTASE™ сell detachment solution (Stem Cell Technologies) and Rock-inhibitor Y-27632 (5 μM; Abcam), the plastic surface being pre-coated with Matrigel solution (1/40 in DMEM/F12) (Corning). The cells were cultured in mTeSR™1 medium (Stem Cell Technologies) at 37°C in a CO₂-incubator with 5% CO₂ and 100% humidity.

Galiakberova A.A., Brovkina O.I., Kondratyev N.V., Artyuhov A.S., Momotyuk E.D., Kulmukhametova O.N., Lagunin A.A., Shilov B.V., Zadorozhny A.D., Zakharov I.S., Okorokova L.S., Golimbet V.E, & Dashinimaev E.B. (2023). Different iPSC-derived neural stem cells shows various spectrums of spontaneous differentiation during long term cultivation. Frontiers in Molecular Neuroscience, 16, 1037902.

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Differentiation of iPSCs to NPCs

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A control iPSC line KYOU-DXR0109B (201B7) was obtained from ATCC and grown in CellMatrix basement membrane gel (ACS-3035) and Pluripo-tent Stem Cell SFM XF/FF media (ACS-3002) at 37°C and 5% CO₂. The iPSC line (ND34391*H) with α-Syn SNCA gene triplication (SNCA-tri) was obtained from the CORIELL Institute cell repository. SNCA-tri iPSCs were initially grown in 0.1% gelatin-coated 6-well plates covered with γ-irradiated CF-1 mouse embryonic fibroblasts and DMEM/F12 20% knock-out serum replacement (Gibco 11330–032, 10828010). At passage 3, SNCA-tri iPSCs were transitioned into a feeder-free system and maintained in a CellMatrix-coated dish and Essential 8 medium (E8M; Gibco A1517001). Derivation of NPCs from both control and SNCA-tri iPSCs was done using PSC neural induction medium (Gibco A1647801) as per the manufacturer’s protocol. Briefly, E8M was replaced with neural induction medium approximately 24 h after passaging iPSCs, which were maintained in this medium for 7 days. Then the NPCs (P0) were passaged onto Geltrex (Thermo Fisher)-coated 6-well plates and expanded in StemPRO neural stem cell SFM media (A1050901). Neural induction efficiency was determined at passage 3 by immunofluorescence staining with a pluripotent marker (Oct4) and neural lineage stem cell marker (Nestin).

Vasquez V., Mitra J., Hegde P.M., Pandey A., Sengupta S., Mitra S., Rao K.S, & Hegde M.L. (2017). Chromatin-Bound Oxidized α-Synuclein Causes Strand Breaks in Neuronal Genomes in in vitro Models of Parkinson’s Disease. Journal of Alzheimer's disease : JAD, 60(Suppl 1), S133-S150.

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Comprehensive Profiling of Diverse iPSC Lines

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The lines used were: H9 hESC (Wicell), NCRM1 (NIH), KYOU-DXR0109B (ATCC), AIW002-02, AJC001-5, AJG001-C4 (these three lines are from the same donor but reprogrammed using different methods or from different cell types), AJD002-3, TD03 (these two lines are from the same donor but reprogrammed with different methods), TD2, TD10, TD22, 3448, and 3450. The complete profiles of the iPSCs are listed in Table 1. The use of iPSCs and stem cells in this research was approved by the McGill University Health Centre Research Ethics Board (DURCAN_IPSC/2019-5374).

Chen C.X., Abdian N., Maussion G., Thomas R.A., Demirova I., Cai E., Tabatabaei M., Beitel L.K., Karamchandani J., Fon E.A, & Durcan T.M. (2021). A Multistep Workflow to Evaluate Newly Generated iPSCs and Their Ability to Generate Different Cell Types. Methods and Protocols, 4(3), 50.

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Murine, Human Cell Line, and iPSC Differentiation Protocol

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The murine IL-3-dependent 32D hematopoietic cell line, human SH-SY5Y, and human induced pluripotent stem cells (hiPSCs, KYOUDXR0109B) were originally obtained from ATCC (Manassas, VA). 32D cells were maintained in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 U/ml penicillin and 100 μg/ml streptomycin (Atlanta Biologicals, Norcross, GA), and 5 ng/mL recombinant murine IL-3 (PeproTech, Rocky Hill, NJ). For inducing 32D cell differentiation, cells were washed with HBSS to remove IL-3, and 500,000 cells were plated on Day 0 in RPMI 1640 containing 10% FBS, 1% penicillin-streptomycin, and 40 ng/mL recombinant murine G-CSF (PeproTech, Rocky Hill, NJ). At the indicated time points, cells were harvested, cytospun, and then Wright’s-stained (EMD Chemicals) for analysis of nuclear morphology to determine the extent of granulocytic differentiation^{54 (link)}. SHSY5Y human neuroblastoma cells were cultured in DMEM/F12 medium with 10% fetal bovine serum (Thermo Fisher Scientific/Gibco). Human iPSCs were cultured in Essential 8 Medium (Thermo Fisher Scientific/Gibco), as per the supplier's protocol.

MacNicol M.C., Cragle C.E., McDaniel F.K., Hardy L.L., Wang Y., Arumugam K., Rahmatallah Y., Glazko G.V., Wilczynska A., Childs G.V., Zhou D, & MacNicol A.M. (2017). Evasion of regulatory phosphorylation by an alternatively spliced isoform of Musashi2. Scientific Reports, 7, 11503.

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Cell Culture Protocols for Cancer and Stem Cell Research

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The cell lines used were described including the human beast adenocarcinoma cell lines, MDA-MB-231 and MDA-MB-468, human induced pluripotent stem (iPS) cell line, KYOU-DXR0109B (obtained from ATCC) and human embryonic stem (hES) cell, TW1 (obtained from BCRC). Human embryonic kidney 293T cell line was used in transient transfection experiments. MDA-MB-231, MDA-MB-468 and 293T cell lines were cultured in Dubelcco's modified Eagle's medium (Gibco, Thermo Fisher Scientific Inc.) containing 10% fetal bovine serum (FBS) in a 5% CO₂/95% air incubator. iPS and hES cell lines were maintained in mTeSR1 medium (Stemcell Technologies Inc.) on Geltrex (Thermo Fisher Scientific Inc.)-coated plate in a 5% CO₂/95% air incubator.

Chen H.F., Chang C.T., Hsu K.W., Peng P.H., Lai J.C., Hung M.C, & Wu K.J. (2023). Epigenetic regulation of asymmetric cell division by the LIBR-BRD4 axis. Nucleic Acids Research, 52(1), 154-165.

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Comprehensive iPSC Line Profiling

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The lines used were: H9 hESC (Wicell), NCRM1 (NIH), KYOU-DXR0109B (ATCC), AIW002-02, AJC001-5, AJG001-C4 (same donor but reprogrammed using different methods or from different cell types), AJD002-3, TD03 (same donor but reprogrammed with different methods), TD2, TD10, TD22, 3448 and 3450. The complete profiles of the iPSCs are listed in

Chen C.X., Abdian N., Maussion G., Thomas R.A., Demirova I., Cai E., Tabatabaei M., Beitel L.K., Karamchandani J., Fon E.A., & Durcan T.M. (2021). Standardized quality control workflow to evaluate the reproducibility and differentiation potential of human iPSCs into neurons.

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