Ldn193189

Manufactured by Merck Group

Sourced in United States, United Kingdom, Germany

LDN193189 is a small molecule inhibitor of the ALK2 (ACVR1) receptor. It functions by blocking the kinase activity of ALK2, a type I bone morphogenetic protein (BMP) receptor. This inhibition can modulate BMP signaling pathways.

Automatically generated - may contain errors

Lab products found in correlation

90 protocols using ldn193189

Modeling Chondrocyte Degeneration with LPS

Check if the same lab product or an alternative is used in the 5 most similar protocols

To establish the chondrocyte degeneration model, chondrocytes were stimulated with 1 µg/ml LPS (Sigma-Aldrich; Merck KGaA) for 24 h (LPS group) at 37°C. Then, in order to examine the relationship of the TIMP3/TGF-β1 signaling pathway and cartilage degeneration, chondrocytes treated with LDN-193189 (0.5 µM; Sigma-Aldrich; Merck KGaA), an inhibitor of the TGF-β1 signaling pathway, for 24 h (LDN-193189 group) at 37°C. The chondrocytes without treatment were used as the control group.

Zhao D.L., Li H.T, & Liu S.H. (2020). TIMP3/TGF-β1 axis regulates mechanical loading-induced chondrocyte degeneration and angiogenesis. Molecular Medicine Reports, 22(4), 2637-2644.

+ Open protocol

+ Expand

Generation of Oligocortical Spheroids from hESCs

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human embryonic stem cells (line H7, WiCell) were grown in mTesR1 media and oligocortical spheroids were generated with minor modifications to the protocol previously described (Madhavan et al., 2018 (link)). Briefly, in the first step of generating oligocortical spheroids, CloneR (Stem Cell Technologies, 5889) was used instead of Y-27632 and Dorsomorphin was replaced with 150nM LDN193189 (Sigma, SML0559). Spheroids were treated with 150nM LDN193189 and 10μM SB-43152 (Sigma, S4317) for the first 6 days followed 20ng/ml FGF-2 (R&D Systems, 233-FB-25/CF) and 20ng/ml EGF (R&D Systems, 236-EG-200) from day 7 to 25.
This was followed by 10ng/ml BDNF (R&D Systems, 248-BD) and 20ng/ml NT-3 (R&D Systems, 267-N3) treatment every other day between days 27 and 40. For OPC development and oligodendrocyte differentiation cultures 10ng/ml PDGF-AA (R&D Systems, 221-AA) and 10ng/ml IGF (R&D Systems, 291-GF-200) were added to cultures every other day between days 51 and 60 an 40n/ml T3 (Sigma, T6397) every other day between days 61 and 70. Spheroids were treated every other day with vehicle DMSO or 300nM MEKi between days 70 and 74 and harvested on day 90. Spheroids were treated with 200μM Hypoxyprobe-1 two hours prior to harvesting for IHC (pimonidazole, Hypoxyprobe Inc, Burlington MA, HP1–100Kit).

Allan K.C., Hu L.R., Scavuzzo M.A., Morton A.R., Gevorgyan A.S., Cohn E.F., Clayton B.L., Bederman I.R., Hung S., Bartels C.F., Madhavan M, & Tesar P.J. (2020). Non-Canonical Targets of HIF1a Impair Oligodendrocyte Progenitor Cell Function. Cell stem cell, 28(2), 257-272.e11.

+ Open protocol

+ Expand

Midbrain Neuron Differentiation from hiPSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

On day −1, 400,000 KOLF2-1J or SFC065 (kindly provided by the laboratory of C. Klein) hiPSC/cm² were seeded in Matrigel-coated 6-well plate wells in StemFlex medium supplemented with 10 μM RI. On day 0, medium was switched to Neurobasal containing 0.5x B27 supplement without vitamin A, 0.5x N2, GlutaMAX, Pen/strep, non-essential amino acids, and LDN193189 (500 nM, Sigma), SB431542 (10 μM, Tocris), SHH-C24II (200 ng/ml, Miltenyi Biotec), Purmorphamine (0.7 μM, Sigma) and 0.7 μM CHIR99021 (Stemcell Technologies). CHIR99021 concentration was raised to 3 μM from day 4 to day 11, moment at which it was withdrawn from the medium. LDN193189 (500 nM, Sigma), SB431542 (10 μM, Tocris), SHH-C24II (200 ng/ml, Miltenyi Biotec), Purmorphamine (0.7 μM, Sigma) were withdrawn from the medium at day 7 and FGF8b (100 ng/mL, R&D Systems) was introduced from day 9 until day 16. At day 11, medium was shifted to Neurobasal-A medium (Life Technologies; 10888-022) supplemented with 1x B27 without Vit. A (Life Technologies; 12587010), 1x GlutaMAX (Life Technologies; 35050-038), 1x PenStrep (Life Technologies; 15140-122), 10 ng/mL BDNF (R&D Systems; 248-BDB-050/CF), 10 ng/mL GDNF (R&D Systems; 212-GD-010), 200 μM ascorbic acid, 0.5 mM dbcAMP (Sigma-Aldrich), 10 μM DAPT (Tocris; 2634). On day 17, ventral midbrain neural progenitors were cryopreserved.

Pantazis C.B., Yang A., Lara E., McDonough J.A., Blauwendraat C., Peng L., Oguro H., Kanaujiya J., Zou J., Sebesta D., Pratt G., Cross E., Blockwick J., Buxton P., Kinner-Bibeau L., Medura C., Tompkins C., Hughes S., Santiana M., Faghri F., Nalls M.A., Vitale D., Ballard S., Qi Y.A., Ramos D.M., Anderson K.M., Stadler J., Narayan P., Papademetriou J., Reilly L., Nelson M.P., Aggarwal S., Rosen L.U., Kirwan P., Pisupati V., Coon S.L., Scholz S.W., Priebe T., Öttl M., Dong J., Meijer M., Janssen L.J., Lourenco V.S., van der Kant R., Crusius D., Paquet D., Raulin A.C., Bu G., Held A., Wainger B.J., Gabriele R.M., Casey J.M., Wray S., Abu-Bonsrah D., Parish C.L., Beccari M.S., Cleveland D.W., Li E., Rose I.V., Kampmann M., Aristoy C.C., Verstreken P., Heinrich L., Chen M.Y., Schüle B., Dou D., Holzbaur E.L., Zanellati M.C., Basundra R., Deshmukh M., Cohen S., Khanna R., Raman M., Nevin Z.S., Matia M., Van Lent J., Timmerman V., Conklin B.R., Chase K.J., Zhang K., Funes S., Bosco D.A., Erlebach L., Welzer M., Kronenberg-Versteeg D., Lyu G., Arenas E., Coccia E., Sarrafha L., Ahfeldt T., Marioni J.C., Skarnes W.C., Cookson M.R., Ward M.E, & Merkle F.T. (2022). A reference human induced pluripotent stem cell line for large-scale collaborative studies. Cell stem cell, 29(12), 1685-1702.e22.

+ Open protocol

+ Expand

Differentiation of Induced Pluripotent Stem Cells into Neural Lineages

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The cells at a density of 1 × 10⁵ cells/cm² were seeded on Matrigel (BD Biosciences) coated plates with Sox2-inducing medium [31 (link)] containing the Optimem 20 ng/mL bFG2, 20 ng/mL EGF, 2 μM SB43152, 1 μM LDN-193189 (Sigma), 1× N2, 1× B27, and 10 ng/mL Leukemia Inhibitory Factor (LIF; Peprotech). On the third day, the medium was switched to neural induction medium [29 (link)] containing the Neurobasal-A medium (Thermo Fisher Scientific) supplemented with 1× N2 supplement (Thermo Fisher Scientific), 20 ng/mL bFG2, 10 ng/mL Brain Derived Neurotrophic Factor (BDNF, Peprotech), 10 ng/mL Glial-Derived Neurotrophic Factor (GDNF, Peprotech), 10 ng/mL Nerve growth factor (NGF, Peprotech), 10 ng/mL Neurotrophin-3 (NT-3, Gibco), 200 μM ascorbic acid (Sigma-Aldrich), 10 μM forskolin (Sigma-Aldrich), 250 μM 3-Isobutyl-1-methylxanthine (IBMX, Sigma-Aldrich) 2 μM SB43152(Sigma-Aldrich), and 1 μM LDN-193189(Sigma-Aldrich).

Gazarian K.G, & Ramírez-García L.R. (2017). Human Deciduous Teeth Stem Cells (SHED) Display Neural Crest Signature Characters. PLoS ONE, 12(1), e0170321.

+ Open protocol

+ Expand

Directed Differentiation of iPSC into Neural Lineages

Check if the same lab product or an alternative is used in the 5 most similar protocols

Undifferentiated iPSK3 cells (2 × 10⁵ cells) were seeded into low attachment 24-well plates in neural differentiation medium composed of DMEM/F-12 plus 2% B27. Y27632 (10 µM) was added during the seeding and removed after 24 h. For dorsal differentiation, the aggregates were treated with 10 µM SB431542 (Sigma) and 100 nM LDN193189 (Sigma) for 7 days. Then the spheroids were treated with FGF2 at 25 ng/mL for another 7 days^{46 (link),47 (link),49 (link)}. To generate a ventral identity, the aggregates were treated with 10 µM SB431542 (Sigma), 100 nM LDN193189 (Sigma), and 5 µM IWP4 (Sigma) for 7 days. Then the spheroids were incubated with 5 µM IWP4 and 1 µM purmorphamine (Sigma) for another 7 days. For maturation, the spheroids were maintained in neural differentiation medium without growth factors for additional 14–38 days (total up to 52 days). The dorsal and ventral identity was characterized using histology, flow cytometry, and RT-PCR.

Song L., Yuan X., Jones Z., Vied C., Miao Y., Marzano M., Hua T., Sang Q.X., Guan J., Ma T., Zhou Y, & Li Y. (2019). Functionalization of Brain Region-specific Spheroids with Isogenic Microglia-like Cells. Scientific Reports, 9, 11055.

+ Open protocol

+ Expand

Colonic Fibroblast and Intestinal Organoid Stimulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

The colonic fibroblast cell line CCD-18co was purchased from ATCC (American Type Culture Collection, Manassas, VA). 18co cells were cultured in DMEM/F12 GlutaMax (Gibco, Thermo Fisher Scientific, Bleiswijk, The Netherlands). Cell growth medium was supplemented with penicillin (100 U/ml, Gibco), streptomycin (100 µg/ml, Gibco), and 10% FCS (Gibco) unless stated otherwise. Fibroblasts were seeded in six well plates and stimulated for up to 96 h with 100 ng recombinant BMP2 (R&D Systems, Abingdon, UK), 200 nM LDN-193189 (Merck), or 100 ng recombinant Noggin (Peprotech, Rocky Hill, NJ, USA) in DMEM/F12. Normal intestinal mouse organoids were stimulated with 50 ng recombinant CXCL12 (Biolegend, Uithoorn, The Netherlands) and monitored for 5 consecutive days.

Ouahoud S., Westendorp B.F., Voorneveld P.W., Abudukelimu S., Koelink P.J., Pascual Garcia E., Buuren J.F., Harryvan T.J., Lenos K.J., van Wezel T., Offerhaus J.A., Fariña-Sarasqueta A., Crobach S., Slingerland M., Hardwick J.C, & Hawinkels L.J. (2022). Loss of bone morphogenetic protein signaling in fibroblasts results in CXCL12-driven serrated polyp development. Journal of Gastroenterology, 58(1), 25-43.

+ Open protocol

+ Expand

Myogenic Differentiation of MSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Myogenic differentiation was induced by culturing MSCs in tissue culture plates coated with 0.01% Collagen Type 1 (Merck cat # C8919) and 20 ug/ml Laminin (Merck, cat # L2020) at a density of 10,000 cells/cm² in growth medium. After reaching a confluence of 70%, myogenic differentiation was induced by adding M1 medium (DMEM, 5% horse serum, 0.1 µM dexamethasone, and 50 µM hydrocortisone) or M2 medium (DMEM, 15% KnockOut Serum Replacement (KSR, ThermoFisher Scientific), 0.5% DMSO (Merck), CHIRON99021 (Tocris) at 1 μM and 0.1 μM LDN193189 (Tocris) for 5 days followed by DMEM, 15% KSR, 0.1% BSA supplemented with 10 ng/ml HGF, 2 ng/ml IGF-1, 20 ng/ml FGF-2 (Peprotech, Merck) and 0.1 μM LDN193189 for the rest of the incubation period. For comparative analysis of myogenic differentiation between M1 and M2 media, media was changed every other day. We performed 3 technical replicates for each of the 5 biological donors for UCB and UCT.

Mishra S., Sevak J.K., Das A., Arimbasseri G.A., Bhatnagar S, & Gopinath S.D. (2020). Umbilical cord tissue is a robust source for mesenchymal stem cells with enhanced myogenic differentiation potential compared to cord blood. Scientific Reports, 10, 18978.

+ Open protocol

+ Expand

APAP-Induced Injury and BMP Receptor Inhibitors

Check if the same lab product or an alternative is used in the 5 most similar protocols

For specific approaches, stimulation with APAP was performed in presence of different pharmacological inhibitors of BMP type I receptors ALK2 and ALK3: LDN-193189, DMH2 and ML347 (SML-0559, Merck Life Science; 5580 and 4945, Tocris Bioscience, Minneapolis, USA). Inhibitors’ affinity for each receptor and working concentration is shown in Table 1. Inhibitors were used as pre-treatment 1 h prior to APAP stimulation or 1 h after of APAP challenge.Table 1

Pharmacological inhibitors of BMP type I receptors ALK2 and ALK3.

Table 1

	Ki ALK2 (nM)	Ki ALK3 (nM)	Working concentration	Control
LDN-193189	5	30	500 nM	–
DMH2	43	5.4	5 or 10 μM	DMSO
ML347	32	>200 fold selectivity	150 nM	DMSO

BMP, Bone morphogenetic protein.

Marañón P., Rey E., Isaza S.C., Wu H., Rada P., Choya-Foces C., Martínez-Ruiz A., Martín M.Á., Ramos S., García-Monzón C., Cubero F.J., Valverde Á.M, & González-Rodríguez Á. (2024). Inhibition of ALK3-mediated signalling pathway protects against acetaminophen-induced liver injury. Redox Biology, 71, 103088.

+ Open protocol

+ Expand

Ectodermal Differentiation of Mouse ESCs

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse embryonic stem cells (129/Sv) were cultured in ESC medium (LIF+2i) (DMEM high glucose with GlutaMAX-1 [Gibco], 20% FBS [Gibco], 1 × non-essential amino acid mix [Gibco], 0.05 mM β-mercaptoethanol, 10 µg/ml LIF [Sigma], 3 µM CHIR99021, 1 µM PD0325901 [both Axon Medchem]) in gelatine-coated culture dishes at 37 °C and 5% CO₂. Induction of embryoid body (EB) formation and EB outgrowth were performed as previously described [21 (link)]. Differentiation of ESCs into the ectodermal lineage was performed as described previously [22 ]. In brief, ESCs were cultured in N2B27 supplemented serum-free medium, containing 10 µg/mL LIF, 3 µM CHIR99021, and 1 µM PD0325901 for 24 h in 25 cm² flasks before passaging to 6-well plates in the same medium containing only 0.4 µM PD0325901 for 2 days. After that, cells were incubated with 1 µM of LDN193189 (BMP antagonist; Sigma) for additional 4 days.

Trixl L., Amort T., Wille A., Zinni M., Ebner S., Hechenberger C., Eichin F., Gabriel H., Schoberleitner I., Huang A., Piatti P., Nat R., Troppmair J, & Lusser A. (2017). RNA cytosine methyltransferase Nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity. Cellular and Molecular Life Sciences, 75(8), 1483-1497.

+ Open protocol

+ Expand

Neuroepithelial Differentiation from hESCs

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Neuroepithelium differentiation to primarily central nervous system epithelia requires the single-cell passaging of hESCs and the culturing of these cells to nearly 100% confluence on Matrigel-coated plates prior to beginning neural induction [19 (link), 20 (link)]. Upon reaching 100% confluence, the MEF-CM medium is replaced with N2/B27 Neural Induction medium supplemented with the small molecules SB431542 (Tocris Biosciences) and LDN193189 (Sigma). N2/B27 Neural Induction medium consists of DMEM/F12 (Gibco) as the base, N2 supplement (Invitrogen), B27 supplement minus vitamin A (Invitrogen), L-glutamine (2 mM), penicillin/streptomycin, beta-mercaptoethanol (Sigma), and MEM nonessential amino acids (Sigma). The supplemented small molecules inhibit BMP and TGF-β signalling which is paramount to successful neural induction. The medium is changed daily with a daily addition of the small molecules to final concentrations of 10 μM and 200 nM, respectively (Supplemental Figure S1). The focus for this study is the day 5 intermediate stage and the day 10 neuroepithelium. Further culturing of these cells takes place in the N2/B27 Neural Induction media without factors.

Worley K.E., Chin A.S, & Wan L.Q. (2018). Lineage-Specific Chiral Biases of Human Embryonic Stem Cells during Differentiation. Stem Cells International, 2018, 1848605.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!