Human h9 escs

Manufactured by WiCell

Sourced in United States

The Human H9 ESCs are a well-characterized and widely used line of human embryonic stem cells. They are derived from the inner cell mass of a human blastocyst and have the capacity for self-renewal and pluripotency, which allows them to differentiate into various cell types.

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Lab products found in correlation

Tesr e8 medium, by STEMCELL (3 mentions) Matrigel coated culture dishes, by BD (2 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Mccoy s 5a media, by Thermo Fisher Scientific (1 mentions) Mycoalert mycoplasma detection kit, by Lonza (1 mentions) Essential 8 medium, by Thermo Fisher Scientific (1 mentions) Geltrex, by Thermo Fisher Scientific (1 mentions) Fixation permeabilization solution kit, by BD (1 mentions) Attune nxt flow cytometer, by Thermo Fisher Scientific (1 mentions) Y 27632, by Merck Group (1 mentions) Ethylene diamine tetraacetic acid (edta), by Thermo Fisher Scientific (1 mentions) Tesr e8 medium, by Merck Group (1 mentions)

5 protocols using human h9 escs

Maintaining Human Cell Lines for Research

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Human H9 ESCs (Lot No.: WIC-WA09-MB-001, WiCell, Wisconsin) and its derivatives were maintained at 37 °C, 5% CO2 in chemical defined medium TeSR-E8 medium (Stemcell Tech.) with 100U/ml penicillin & 100 μg/ml streptomycin (Gibco) on matrigel (#CB40230A, Corning) coated tissue culture vessels. Authentication of H9 ESCs were performed by WiCell. ESCs were passaged every 4 to 6 days to maintain sub-confluence using 0.5 mM EDTA as described previously (20 (link)). Human colon cancer RKO cells (kindly given by Dr. Bert Vogelstein) and its derivatives were maintained at 37 °C, 5% CO2 in McCoy’s 5A media (Fisher) supplemented with 10% FBS and 100U/ml penicillin & 100 μg/ml streptomycin (Gibco). RKO cells were passaged every 3 to 4 days to maintain sub-confluence (Authentication of RKO cell line was performed by JHU-GRCF Biorepository & Cell Center). Cells were screened for mycoplasma before experiments using a MycoAlert™ Mycoplasma Detection Kit (Lonza).
All cell lines were passaged in our laboratory for no more than 30 passages after resuscitation.

Liu C., Banister C.E, & Buckhaults P.J. (2019). Spindle assembly checkpoint inhibition can resensitize p53-null stem cells to cancer chemotherapy. Cancer research, 79(9), 2392-2403.

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Culturing Human Pluripotent Stem Cells

Cited 1 time

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Human H9 ESCs (WiCell, Madison, WI, US, passage 30–50) or human iPSC 317^{31 (link)} were maintained on Geltrex (Thermo Fisher, Waltham, MA, USA) in Essential 8 Medium (Thermo Fisher, Waltham, MA, USA) with daily media change. ROCK inhibitor (Y27632, 10uM, ApexBio, Boston, MA, USA) was added to the culture for 24 hours after passaging to prevent single cell-induced cell death of hPSCs.

Potocnik A.J., Kohler H, & Eichmann K. (1997). Hemato-lymphoid in vivo reconstitution potential of subpopulations derived from in vitro differentiated embryonic stem cells. Proceedings of the National Academy of Sciences of the United States of America, 94(19), 10295-10300.

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Pluripotency and Surface Marker Analysis

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Human H9 ESCs (WiCell, WA09) were used as the positive control for FACS analysis to detect the pluripotency marker OCT4 and the human ESC cell surface marker SSEA4. HEK293T cells were used as the negative cell control for iPSC and NPC marker detection. Cells were dissociated and passed through a 70 µm cell strainer to make single cell suspension. For cell surface marker staining, cells were directly incubated with fluorophore‐conjugated primary antibodies for 20 min on ice. The same fluorophore‐conjugated IgGs were included as the isotype controls. For intracellular OCT4 staining, cells were first fixed and permeabilized using a Fixation/Permeabilization Solution Kit (BD, 554 714) before incubation with the PE‐conjugated anti‐Oct3/4 primary antibody. The PE‐conjugated mouse IgG1 was included as the isotype control. Cells were washed twice and resuspended in phosphate‐buffered saline (PBS) containing DAPI and 0.1% donkey serum. The samples were run on Attune NxT Flow Cytometer (ThermoFisher Scientific) and data were analyzed by FlowJo v10. The detailed information of all the primary antibodies and isotype controls used were listed in Table S7 (Supporting Information).

Feng L., Chao J., Tian E., Li L., Ye P., Zhang M., Chen X., Cui Q., Sun G., Zhou T., Felix G., Qin Y., Li W., Meza E.D., Klein J., Ghoda L., Hu W., Luo Y., Dang W., Hsu D., Gold J., Goldman S.A., Matalon R, & Shi Y. (2020). Cell‐Based Therapy for Canavan Disease Using Human iPSC‐Derived NPCs and OPCs. Advanced Science, 7(23), 2002155.

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Hypoxic Retinal Precursor Cells Co-Cultured with Neural Progenitor Cells

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Immortalized R28 retinal precursor cells were cultured as previously described [9 (link)]. H9 human ESCs (WiCell Research Institute, Madison, WI, USA) were routinely maintained on Matrigel-coated culture dishes (BD Biosciences, San Jose, CA, USA) in TeSR™-E8™ medium (STEMCELL Technologies, Vancouver, BC, Canada). The culture process for NPCs is detailed in a recent publication [8 (link)]. A hypoxic environment was created in R28 cells using cobalt chloride (CoCl₂). R28 cells were seeded at a density of 2 × 10⁵ and then treated with 300 µM CoCl₂ for 3 h. Following this, NPCs were added to the damaged R28 cells. After 24 h, the cells were harvested and prepared for analysis.

Shin H.A., Park M., Lee H.J., Duong V.A., Kim H.M., Hwang D.Y., Lee H, & Lew H. (2023). Unveiling Neuroprotection and Regeneration Mechanisms in Optic Nerve Injury: Insight from Neural Progenitor Cell Therapy with Focus on Vps35 and Syntaxin12. Cells, 12(19), 2412.

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Maintaining H9 Human Embryonic Stem Cells

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H9 human ESCs (WiCell Research Institute, Madison, WI, USA) were routinely maintained on Matrigel-coated culture dishes (BD Biosciences, San Jose, CA, USA) in TeSR™-E8™ medium (STEMCELL Technologies, Vancouver, BC, Canada). For passaging, ESCs were incubated with 0.5 mM EDTA (Thermo Fisher Scientific, Waltham, MA, USA) in a 37 °C CO₂ incubator for 3 min and then were split in the ratio of 1:20 onto Matrigel-coated dishes with TeSR™-E8™ medium containing 10 μM Y-27632 (Sigma-Aldrich, St. Louis, MO, USA). TeSR™-E8™ medium without Y-27632 was changed daily from two days after passage. Experiments using hESCs were approved by the Institutional Review Board of CHA University (IRB No. 1044308-201603-LR-004-09).

Park M., Kim H.M., Shin H.A., Lee S.H., Hwang D.Y, & Lew H. (2021). Human Pluripotent Stem Cell-Derived Neural Progenitor Cells Promote Retinal Ganglion Cell Survival and Axon Recovery in an Optic Nerve Compression Animal Model. International Journal of Molecular Sciences, 22(22), 12529.

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