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Ldn 193189 dihydrochloride

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LDN-193189 dihydrochloride is a potent and selective small-molecule inhibitor of the bone morphogenetic protein (BMP) type I receptors ALK2, ALK3, and ALK6. It functions by blocking BMP signaling, which is involved in various cellular processes.

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Lab products found in correlation

Ecis 1600r, by Applied Biophysics (2 mentions) Forskolin, by Bio-Techne (1 mentions) Ascorbic acid, by Bio-Techne (1 mentions) Lipid supplement, by Thermo Fisher Scientific (1 mentions) Pdgf aa, by R&D Systems (1 mentions) Jagged1, by R&D Systems (1 mentions) Phorbol 12 myristate 13 acetate, by Bio-Techne (1 mentions) Dll 1, by R&D Systems (1 mentions) Jagged1, by Thermo Fisher Scientific (1 mentions) B27 without vitamin a, by Thermo Fisher Scientific (1 mentions) Oncostatin m, by R&D Systems (1 mentions) Dorsomorphin dihydrochloride, by Bio-Techne (1 mentions) Recombinant human bmp 2, by R&D Systems (1 mentions)

4 protocols using ldn 193189 dihydrochloride

1

Astrocyte Differentiation Media Compositions

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Astro-1 medium was composed of DMEM/F12 medium supplemented with N2 (Gibco, 17502048), B27 without vitamin A (Gibco, 12587010), 100 nM LDN-193189 dihydrochloride (Tocris, 6053), and the human recombinant proteins PDGF-AA (R&D Systems, 221-AA), JAGGED-1 (R&D Systems, 1277-JG), DLL-1 (R&D Systems, 1818-DL), ONCOSTATIN M (R&D Systems, 295-OM), LIF (R&D Systems, 7734-LF), and CNTF (R&D Systems, 257-NT) (all at 10 ng/mL concentration).
Astro-2 medium was composed of DMEM/F12 base medium supplemented with N2 (Gibco), B27 complete (Gibco, 17504044), 1% lipid supplement (Gibco, 11905031), and the recombinant proteins JAGGED1, DLL-1, ONCOSTATIN M, LIF, and CNTF (all at 10 ng/mL concentration) (R&D Systems).
Astro-3 medium was composed of DMEM/F12 base medium supplemented with N2, B27 with vitamin A, 1% lipid supplement (Gibco), and JAGGED1, DLL-1, LIF, CNTF (all at 10 ng/mL concentration), hNRG1/EGF domain (20 ng/mL, R&D Systems, 396-HB), 2 μM forskolin (Tocris, 1099), 200 nM phorbol-12 myristate-13 acetate (Tocris, 1201), 40 ng/mL triiodothyronine T3 (Tocris, 6666), and 200 μM ascorbic acid (Tocris, 4055).
Jovanovic V.M., Weber C., Slamecka J., Ryu S., Chu P.H., Sen C., Inman J., De Sousa J.F., Barnaeva E., Hirst M., Galbraith D., Ormanoglu P., Jethmalani Y., Mercado J.C., Michael S., Ward M.E., Simeonov A., Voss T.C., Tristan C.A, & Singeç I. (2023). A defined roadmap of radial glia and astrocyte differentiation from human pluripotent stem cells. Stem Cell Reports, 18(8), 1701-1720.
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2

Examining Sema3A-BMP2 Pathway Interaction

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To determine whether Sema3A interacts with the BMP2 pathway, cultures were treated with chemicals known to inhibit receptors of BMPs. 0.1μM or 1μM of two chemicals known to inhibit ALK2, ALK3, and ALK6 [DMH-2 and dorsomorphin dihydrochloride (DDCl); Tocris Bioscience]; or LDN 193189 dihydrochloride (ALK2 and ALK3 inhibitor; Tocris Bioscience) were used to treat surface cultured MSCs. 0.1μM, 1μM, or 10μM DMH-1 (ALK2 inhibitor; Tocris Bioscience) with or without the addition of 1μg/mL Sema3A was also used to treat surface cultured MSCs. The production of Sema3A after the addition of 40ng/mL recombinant human BMP2 (R&D Systems) was also measured. Cells were plated as described above (Section 2.2), and media were changed every 48h with treatment for 7d. At 7d, cells were incubated with fresh CCM without treatment for 24h and assayed as described previously (Section 2.5).
Lotz E.M., Berger M.B., Boyan B.D, & Schwartz Z. (2020). Regulation of Mesenchymal Stem Cell Differentiation on Microstructured Titanium Surfaces by Semaphorin 3A. Bone, 134, 115260.
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3

Modeling Brain Barrier Integrity with bEnd.3 Cells

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Murine bEnd.3 cell line (ATCC, CRL-229) was isolated from brain tissue derived from BALB/c mouse with endothelioma and was is sub-cultured at a 1:3 to 1:4 split ratio approximately every 3 days. These cells were widely used in both monolayer and in co-culture with astrocytes and pericytes in transwells to establish BBB models and were both established as valid in vitro models based on the barrier integrity as determined by the transendothelial electrical resistance/TEER [29 (link)]. We have compared the bEnd.3 monolayer with the eEnd.3 and C8-D1A (astrocyte type I clone, ATCC, CRL-2541) co-culture (with either bEnd3 on top or on bottom in transwells) and repeated the experiments in both systems (N = 3–4 on each assay of Western blots and TEER measurement). Cells were treated with murine recombinant rBMP4 (R&D 5020-BP; 25 ng/mL) for various time points with or without adding BMP4 inhibitor (LDN 193189 dihydrochloride, 100 nM, Bio-Techne Corporation). TEER was determined using the device model (ECIS 1600R, Applied Biophysics, Troy, NY).
Chen X., Chen L., Lin G., Wang Z., Kodali M.C., Li M., Chen H., Lebovitz S.G., Ortyl T.C., Li L., Ismael S., Singh P., Malik K.U., Ishrat T., Zhou F.M., Zheng W, & Liao F.F. (2022). White matter damage as a consequence of vascular dysfunction in a spontaneous mouse model of chronic mild chronic hypoperfusion with eNOS deficiency. Molecular Psychiatry, 27(11), 4754-4769.
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4

Establishing Murine bEnd.3 BBB Models

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Murine bEnd.3 cell line (ATCC, CRL-229) was isolated from brain tissue derived from BALB/c mouse with endothelioma and was is sub-cultured at a 1:3 to 1:4 split ratio approximately every 3 days. These cells were widely used in both monolayer and in co-culture with astrocytes and pericytes in transwells to establish BBB models and were both established as valid in vitro models based on the barrier integrity as determined by the transendothelial electrical resistance/TEER [29 (link)]. We have compared the bEnd.3 monolayer with the eEnd.3 and C8-D1A (astrocyte type I clone, ATCC, CRL-2541) co-culture (with either bEnd3 on top or on bottom in transwells) and repeated the experiments in both systems (N = 3–4 on each assay of Western blots and TEER measurement). Cells were treated with murine recombinant rBMP4 (R&D 5020-BP; 25 ng/mL) for various time points with or without adding BMP4 inhibitor (LDN 193189 dihydrochloride, 100 nM, Bio-Techne Corporation). TEER was determined using the device model (ECIS 1600R, Applied Biophysics, Troy, NY).
Chen X., Chen L., Lin G., Wang Z., Kodali M.C., Li M., Chen H., Lebovitz S.G., Ortyl T.C., Li L., Ismael S., Singh P., Malik K.U., Ishrat T., Zhou F.M., Zheng W, & Liao F.F. (2022). White matter damage as a consequence of vascular dysfunction in a spontaneous mouse model of chronic mild chronic hypoperfusion with eNOS deficiency. Molecular Psychiatry, 27(11), 4754-4769.
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