Poly l ornithine

Manufactured by Merck Group

Sourced in United States, Germany, Sao Tome and Principe, Japan, Canada, Macao, Switzerland, France

Poly-L-ornithine is a synthetic polymer consisting of the amino acid L-ornithine. It is commonly used as a cell culture substrate to promote cell attachment and growth.

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Lab products found in correlation

Close spelling variants of this product

Poly-ornithine (150 citations) L-ornithine (30 citations) Poly-L-ornithine solution (30 citations) Poly-L-ornithine hydrobromide (27 citations) Poly-L-ornithine (PLO, (24 citations) Poly-L-ornithine/laminin (18 citations) Poly-L-ornithine-coated (8 citations) Poly-L-ornithine and laminin (5 citations) Poly-L-ornithine hydrobromide (PLO) (5 citations) Poly-L-ornithine and fibronectin (3 citations)

475 protocols using poly l ornithine

Spontaneous Differentiation of Oligodendrocyte/Neuron Progenitors

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Example 81

Spontaneous differentiation of human oligodendrocyte/neuron progenitors in 24-well and 96-well plates. Coat plate with 10 μg/ml poly-L-ornithine (Sigma Cat. No. P4957) overnight at room temperature. Remove the poly-L-ornithine solution and rinse the plate with 1×PBS, add 10 μg/ml laminin (Millipore Cat. No. CC095) and incubate at 37° C. for at least 2 hours. Seed cells on the coated plates in expansion media and allowed to attach overnight. The next day expansion media was removed and replaced with freshly prepared spontaneous differentiation media only containing OPC/neuron basal media and N21 media supplement. For 14 day culture, differentiation media was replaced every 3-4 days. Cells were imaged and neurite length was analyzed by IncuCyte S3 Live Cell Analysis System every 4 hours. After 14 days of differentiation, cells were fixed by 4% paraformaldehyde and stained with DAPI and Neuron-specific ß-III tubulin antibody (R&D systems, NL1195R).

US11479533B2. Inhibitors of Rho associated coiled-coil containing protein kinase (2022-10-25). KADMON CORPORATION, LLC [US]. Inventors: Eduardas Skucas [US], Kevin G. Liu [US], Ji-In Kim [US], Masha V. Poyurovsky [US], Rigen Mo [US], Jingya Zhang [US].

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Isolation and Differentiation of Mouse NSCs

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NSCs were isolated from E12.5 mouse embryonic cortex, in accordance with a previously described procedure [28 (link), 29 (link)]. The primary NSCs were seeded onto poly-l-ornithine (Sigma-Aldrich)- and laminin (Invitrogen)-coated dishes, cultured as monocultures. Neural basal medium with EGF (20 ng/ml; PeproTech), bFGF (20 ng/ml; PeproTech), heparin (20 ng/ml; Sigma-Aldrich), and 2% B27 (Invitrogen) was used as NSC proliferation medium. For differentiation, NSCs were seeded on poly-l-ornithine (Sigma-Aldrich)- and laminin (Invitrogen)-coated dishes. Upon NSC attachment, the medium was changed to differentiation medium identical to the NSC proliferation medium without growth factors (EGF and bFGF). After differentiation for 10 days, neural- and glial-specific markers were used to determine the differentiation potential of cultured NSCs.

Tian G.G., Zhao X., Hou C., Xie W., Li X., Wang Y., Wang L., Li H., Zhao X., Li J, & Wu J. (2022). Integrative analysis of the 3D genome structure reveals that CTCF maintains the properties of mouse female germline stem cells. Cellular and Molecular Life Sciences, 79(1), 22.

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Astrocyte-Neuron Co-culture Viability Assay

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Cryopreserved astrocytes from Control and Krabbe donors were seeded onto poly-L-ornithine (10 μg/mL, Sigma-Aldrich) and laminin (10 μg/mL, Sigma-Aldrich) 96 well CellCarrier Ultra high content imaging plates (PerkinElmer) in complete astrocyte differentiation medium at 10,000 cells/well. Following 3 weeks of culture, with 50% astrocyte medium replaced twice per week, medium was fully replaced with complete NDM-2 and conditioned for 96 hours. At this time, cryopreserved neurons from the Control 2 donor were seeded at 10,000 cells/well onto a poly-L-ornithine (10 μg/mL, Sigma-Aldrich) and laminin (10 μg/mL, Sigma-Aldrich) 384-well CellCarrier Ultra high content imaging plate (PerkinElmer) in NDM-2. Following 96-hours of culture, astrocyte conditioned medium was collected and medium from the neurons was aspirated and replaced with the astrocyte conditioned medium. After 96-hours of exposure to astrocyte conditioned medium, neuron viability was quantified by incubating with Calcein Green AM, NucRed Dead, and Hoechst fluorescent probes in PBS +magnesium +calcium for 10 minutes at 37C and imaging on a PerkinElmer Opera Phenix microscope. Cells were imaged with a 20X objective and viability was calculated as the percentage of Calcein Green AM positive, NucRed Dead negative nuclei over the total nuclei (identified with Hoechst) using associated Perkin Elmer Harmony software.

Lieberman R., Cortes L.K., Gao G., Park H., Wang B., Jones P.L., Hunter R.B., Leonard J.P, & Barker RH J.r. (2022). Human iPSC-derived astrocytes generated from donors with globoid cell leukodystrophy display phenotypes associated with disease. PLoS ONE, 17(8), e0271360.

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Differentiation and Maintenance of LUHMES Cells

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For proliferation, LUHMES cells were kept in flasks coated with poly-L-ornithine (0.1 mg/mL; 4 °C, overnight; Sigma-Aldrich) in a DMEM/F12 medium (Sigma-Aldrich) with 1% N2-supplement (Thermo Fisher Scientific, Cat#: 17502048), 2 mM L-Glutamine (Thermo Fisher Scientific), and a 0.04 μg/mL human basic fibroblast growth factor (bFGF, Gibco, Cat#: 13256-029). For differentiation, the cells were seeded in a differentiation medium at a density of 100,000 cells per cm² in the cell culture dishes coated with poly-L-ornithine (50 μg/mL; 4 °C, overnight; Sigma-Aldrich), followed by coating with human fibronectin (1 μg/mL, 37 °C, overnight; Sigma-Aldrich). The differentiation medium consisted of a DMEM/F12 medium with 1% N2-supplement, 1 μg/mL tetracycline (Sigma-Aldrich), 1 mM dibutyryl cAMP (Sigma-Aldrich), and a 2 ng/mL glial cell-derived neurotrophic factor (GDNF) (Sigma-Aldrich, Cat#: G1777). To handle LUHMES spheroid culture, between 1000 and 3000 cells were seeded into ultra-low attachment, U-shaped, 96-well plate (CellCarrier Spheroid ULA 96-well Microplates, Perkin Elmer, Cat#: 6055330) with 100 μL of LUHMES proliferating medium.

Vianello C., Dal Bello F., Shin S.H., Schiavon S., Bean C., Magalhães Rebelo A.P., Knedlík T., Esfahani E.N., Costiniti V., Lacruz R.S., Covello G., Munari F., Scolaro T., Viola A., Rampazzo E., Persano L., Zumerle S., Scorrano L., Gianelle A, & Giacomello M. (2023). High-Throughput Microscopy Analysis of Mitochondrial Membrane Potential in 2D and 3D Models. Cells, 12(7), 1089.

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Culturing Lund Human Mesencephalic Cells

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Lund human mesencephalic cells were cultured as described previously (Höllerhage et al., 2017 ). Briefly, cells were plated in T75 flasks (EasYFlasks, Nunclon DELTA, Thermo Fisher Scientific, Waltham, MA, United States) coated with 50 μg/mL poly-L-ornithine (Sigma-Aldrich, St. Louis, MO, United States) in DMEM/F12 growth medium (Sigma-Aldrich) with 1% N2-supplement (Life Technologies, Carlsbad, CA, United States) and 0.04 μg/mL human basic fibroblast growth factor (bFGF; PeproTech, Rocky Hill, CT, United States). Multi-well dishes and flasks (Nunc MicroWell plates, Thermo Fisher Scientific, Waltham, MA, United States) were coated with 50 μg/mL poly-L-ornithine (Sigma-Aldrich) at 37°C overnight and washed three times with phosphate-buffered saline (PBS; LifeTechnologies) followed by coating with 5 μg/mL fibronectin (Sigma-Aldrich) for 24 h in the incubator (37°C, 5% CO₂). For experiments, cells were plated at a density of 110,000 cells/cm² in differentiation medium [DMEM/F12 with 1% N2-supplement, 1 μg/mL tetracycline, 0.49 mg/mL dibutyryl cyclic-AMP (Sigma-Aldrich), and 2 ng/mL human glial cell-derived neurotrophic factor (GDNF; R&D Systems, Minneapolis, MN, United States)]. Cells were routinely tested for mycoplasma contamination.

Findeiss E., Schwarz S.C., Evsyukov V., Rösler T.W., Höllerhage M., Chakroun T., Nykänen N.P., Shen Y., Wurst W., Kohl M., Tost J, & Höglinger G.U. (2021). Comprehensive miRNome-Wide Profiling in a Neuronal Cell Model of Synucleinopathy Implies Involvement of Cell Cycle Genes. Frontiers in Cell and Developmental Biology, 9, 561086.

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Differentiation of LUHMES Cells into Dopaminergic Neurons

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Undifferentiated Lund human mesencephalic (LUHMES) cells^{17 (link)} were expanded on 50 µg/mL poly-l-ornithine (Sigma-Aldrich, St. Louis, MO) coated T75 flasks (EasYFlasks, Nunclon DELTA, VWR, Darmstadt, Germany) in DMEM/F12 (Sigma-Aldrich) supplemented with 1% N2 supplement (Life Technologies, Carlsbad, CA) and 0.04 µg/mL basic fibroblast growth factor (bFGF; PeproTech, Rocky Hill, CT).
For experiments, cells were seeded on either T25 flasks or multi-well-plates (Nunc MicroWell plates, Thermo Fisher Scientific, Waltham, MA) sequentially coated with 50 µg/mL poly-l-ornithine (Sigma-Aldrich) and 5 µg/mL bovine fibronectin (Sigma-Aldrich). To induce differentiation of LUHMES cells into dopaminergic neurons, differentiation medium consisting of DMEM/F12 with 1% N2 supplement, 1 µg/mL tetracycline, 0.49 µg/mL dibutyryl cyclic-AMP (Sigma-Aldrich) and 2 ng/mL glial cell-derived neurotrophic factor (GDNF; R&D Systems, Minneapolis, MN) was used. Cell density was kept at 100,000 cells/cm² across all flasks and well plate formats. Cells were kept at all times in standard cell culture conditions at 37 °C, 5% CO₂, and water-saturated air. Cells were routinely tested for mycoplasma contamination.

Chakroun T., Evsyukov V., Nykänen N.P., Höllerhage M., Schmidt A., Kamp F., Ruf V.C., Wurst W., Rösler T.W, & Höglinger G.U. (2020). Alpha-synuclein fragments trigger distinct aggregation pathways. Cell Death & Disease, 11(2), 84.

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Proliferating and Differentiating LUHMES Cells

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Proliferating Lund human mesencephalic (LUHMES) cells [28 (link)] were expanded in T75 flasks (EasYFlasks, Nunclon DELTA, VWR, Darmstadt, Germany) coated with 50 µg/mL poly-l-ornithine (Sigma-Aldrich, St. Louis, MO). Cells were kept in growth medium consisting of DMEM/F12 (Sigma-Aldrich), supplemented with 1% N2 supplement (Life Technologies, Carlsbad, CA) and 0.04 µg/mL basic fibroblast growth factor (bFGF; PeproTech, Rocky Hill, CT).
For differentiation, cells were seeded on T75 flasks, T25 flasks, or multi-well plates (Nunc MicroWell plates, Thermo Fisher Scientific, Waltham, MA) sequentially coated with 50 µg/mL poly-l-ornithine (Sigma-Aldrich) and 5 µg/mL bovine fibronectin (Sigma-Aldrich). Cells were cultured in differentiation medium consisting of DMEM/F12 with 1% N2 supplement, 1 µg/mL tetracycline, 0.49 µg/mL dibutyryl cyclic-AMP (Sigma-Aldrich), and 2 ng/mL glial cell–derived neurotrophic factor (GDNF; R&D Systems, Minneapolis, MN). Cells were kept in standard cell culture conditions at 37 °C, 5% CO₂, and water-saturated air at all times. Cell density was kept at 100,000 cells/cm² across all flasks and well plate formats.

Höllerhage M., Wolff A., Chakroun T., Evsyukov V., Duan L., Chua O.W., Tang Q., Koeglsperger T, & Höglinger G.U. (2022). Binding Stability of Antibody—α-Synuclein Complexes Predicts the Protective Efficacy of Anti-α-synuclein Antibodies. Molecular Neurobiology, 59(7), 3980-3995.

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LUHMES Cell Proliferation and Differentiation

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For proliferation, LUHMES cells (15 (link)) were kept on flasks (Nunc, Thermo Fisher Scientific, Waltham, MA, USA), coated with poly-L-ornithine (0.1 mg/ml; 4°C, overnight; Sigma-Aldrich, St. Louis, MO, USA) in a DMEM/F12 medium (Sigma-Aldrich, St. Louis, MO, USA) with 1% N2-supplement (Thermo Fisher Scientific) and a 0.04 μg/ml basic fibroblast growth factor (bFGF, PeproTech, Rocky Hill, CT, USA). For differentiation, the cells were seeded in a differentiation medium at a density of 100,000 cells per cm² on cell culture dishes coated with poly-L-ornithine (0.1 mg/ml; 4°C, overnight; Sigma-Aldrich), followed by coating with fibronection (5 μg/ml, 37°C, overnight; Sigma-Aldrich). The differentiation medium consisted of a DMEM/F12 medium with 1% N2-supplement, 1-μg/ml tetracycline (Sigma-Aldrich), 0.49-μg/ml dibutyryl cyclic adenosine monophosphate, and a 2-ng/ml glial cell-derived neurotrophic factor (GDNF, R&D Systems, Minneapolis, MN, USA).

Höllerhage M., Stepath M., Kohl M., Pfeiffer K., Chua O.W., Duan L., Hopfner F., Eisenacher M., Marcus K, & Höglinger G.U. (2022). Transcriptome and Proteome Analysis in LUHMES Cells Overexpressing Alpha-Synuclein. Frontiers in Neurology, 13, 787059.

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Cryopreservation of Differentiated Monkey ESCs

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The NE cells from monkey ESCs were detached with TrypLE Select (Thermo Fisher Scientific) and suspended in Dulbecco’s Modified Eagle Medium (DMEM)/F12 (Thermo Fisher Scientific) supplemented with 20 ng/mL FGF2, 200 ng/mL SHH, 20 ng/mL PDGF-AA (R&D Systems), 1 × B-27 Supplement without vitamin A (Thermo Fisher Scientific), 1 × N-2 Supplement (Thermo Fisher Scientific), 100 units/mL penicillin, and 100 μg/mL streptomycin (Sigma-Aldrich), and were then allowed to grow in suspension as spheres for 44 days. The medium was changed twice weekly. The spheres were treated with 1 × Accutase (Thermo Fisher Scientific) for 5 min and centrifuged. The cell pellet was resuspended with the same medium and dissociated into single cells by pipetting. The cells were transferred to plates coated with 0.01% poly-L-ornithine (Sigma-Aldrich) and 10 μg/mL laminin (Sigma-Aldrich) (poly-L-ornithine/laminin) and cultured for 7 days. The medium was changed twice weekly. The cells were detached with Accutase and centrifuged. The cells were suspended in STEM-CELLBANKER (Takara Bio) and cryopreserved in -80°C refrigerator.

Yamashita T., Miyamoto Y., Bando Y., Ono T., Kobayashi S., Doi A., Araki T., Kato Y., Shirakawa T., Suzuki Y., Yamauchi J., Yoshida S, & Sato N. (2017). Differentiation of oligodendrocyte progenitor cells from dissociated monolayer and feeder-free cultured pluripotent stem cells. PLoS ONE, 12(2), e0171947.

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Induction and Maintenance of EpiLCs from ESCs

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ESCs were cultured in N2B27 basal medium containing 3 μM CHIR99021 (Wako), 0.4 μM PD0325901 (Wako), and LIF in a dish coated with 0.01% poly-L-ornithine (Millipore) and 10 ng/mL laminin (BD Biosciences). Induction of EpiLCs was performed as described previously (Hayashi et al., 2011 (link)). EpiLCs were induced from ESCs in N2B27 basal medium containing 20 ng/mL activin A (Peprotech), 12 ng/mL bFGF (Invitrogen), and 0.1–1% KSR for 2 days on a dish coated with 16.7 μg/mL human plasma fibronectin (Millipore). Two days after EpiLC induction, the cells were collected and replated (0.5–1.0 × 10⁵) with or without doxycycline in GK15 medium on a dish coated with 16.7 μg/mL human plasma fibronectin. For the colony formation assay for AP-positive cells, ESCs, EpiLCs, and EpiLCs induced with or without PRDM14 were dissociated by TrypLE Select (Invitrogen). Cells (1.0 × 10⁴) were cultured under standard ESC culture conditions. After culture for 3 days, the cells were stained for AP activity. The aggregate culture of EpiLCs was performed as previously reported (Nakaki et al., 2013 (link)). In brief, after 36 hr of EpiLC induction the cells were collected, and 2,000 cells per well were transferred to a low-cell binding 96-well plate (NUNC). The cells were grown for 2 days in GK15 medium with or without 1.5 μg/mL doxycycline.

Okashita N., Suwa Y., Nishimura O., Sakashita N., Kadota M., Nagamatsu G., Kawaguchi M., Kashida H., Nakajima A., Tachibana M, & Seki Y. (2016). PRDM14 Drives OCT3/4 Recruitment via Active Demethylation in the Transition from Primed to Naive Pluripotency. Stem Cell Reports, 7(6), 1072-1086.

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