Activin a

Manufactured by Miltenyi Biotec

Sourced in United States, Germany

Activin A is a recombinant human protein that belongs to the transforming growth factor-beta (TGF-β) superfamily. It is a dimeric protein composed of two inhibin beta A subunits. Activin A plays a key role in the regulation of various cellular processes, including cell differentiation, proliferation, and apoptosis.

Automatically generated - may contain errors

Lab products found in correlation

Bone morphogenetic protein 4 (bmp4), by R&D Systems (12 mentions) Matrigel, by Corning (7 mentions) Xav939, by R&D Systems (6 mentions) Chir99021, by Axon Medchem (5 mentions) Retinoic acid, by Merck Group (5 mentions) Fibroblast growth factor 2 (fgf2), by Miltenyi Biotec (3 mentions) Bone morphogenetic protein 4 (bmp4), by Miltenyi Biotec (3 mentions) Stemflex medium, by Thermo Fisher Scientific (3 mentions) Revitacell, by Thermo Fisher Scientific (3 mentions) Bone morphogenetic protein 4 (bmp4), by Merck Group (3 mentions) Sb431542, by R&D Systems (3 mentions) Essential 8 medium, by Thermo Fisher Scientific (3 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Oncostatin m, by Miltenyi Biotec (2 mentions) Hepatocyte culture medium, by Lonza (2 mentions) Hepatocyte growth factor (hgf), by Miltenyi Biotec (2 mentions) Vascular endothelial growth factor (vegf), by R&D Systems (2 mentions) Xav939, by Bio-Techne (2 mentions) Sb431542, by Bio-Techne (2 mentions) Chir99021, by Bio-Techne (2 mentions)

Spelling variants of this product

Human recombinant Activin A (2 citations)

21 protocols using activin a

Stepwise Differentiation of hiPSCs into Mature Hepatocytes

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

Control hiPSCs were differentiated into HLCs as previously described (Si-Tayeb et al., 2016 ) in triplicates. Briefly, once cells reach ∼70%–80% confluency, hiPSCs were cultured in RPMI 1640 medium (Life Technologies) supplemented with B27 (with insulin) (Life Technologies), Activin A 100 ng/mL (Miltenyi), FGF2 20 ng/mL (Miltenyi), and BMP4 10 ng/mL (Miltenyi) for 2 days in normoxia (20% O₂, 5% CO₂), then switched to RPMI 1640 with Activin A 100 ng/mL for 3 days to induce definitive endoderm (DE) cells. DE cells were further differentiated into hepatic progenitor cells for 5 days in RPMI supplemented with BMP4 20 ng/mL and FGF2 10 ng/mL in hypoxia (4% O₂, 5% CO₂). Then cells were cultured for 5 days into immature hepatocytes in RPMI 1640 supplemented with HGF 20 ng/mL (Miltenyi) in hypoxia (4% O₂, 5% CO₂). Then, cells were directed into mature hepatocytes being cultured in hepatocyte culture medium (HCM) (Lonza) supplemented with Oncostatin M 20 ng/mL (Miltenyi) for additional 5–6 days in normoxia (20% O₂, 5% CO₂). RNA samples were collected every day of the differentiation (day 0 to day 20) and processed further for qPCR (detailed explanation is shown in the gene expression analysis). Twenty-four-hour conditioned medium was collected, centrifuged, then kept for further secretion study using an ELISA assay kit.

Roudaut M., Idriss S., Caillaud A., Girardeau A., Rimbert A., Champon B., David A., Lévêque A., Arnaud L., Pichelin M., Prieur X., Prat A., Seidah N.G., Zibara K., Le May C., Cariou B, & Si-Tayeb K. (2021). PCSK9 regulates the NODAL signaling pathway and cellular proliferation in hiPSCs. Stem Cell Reports, 16(12), 2958-2972.

+ Open protocol

+ Expand

Cardiac and Endothelial Induction from hiPSCs

Cited 1 time

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

Endothelial and cardiac cells were induced from hiPSCs in monolayer culture using the CMEC protocol described previously.^{33 (link)} Briefly, hiPSCs were split at a 1:12 ratio and seeded on 6-well plates coated with 75 µg/mL (growth factor reduced) Matrigel (Corning) on day −1. On day 0, cardiac mesoderm was induced by changing TeSR-E8 to BPEL medium,^{36 (link)} supplemented with 20 ng/mL BMP4 (R&D Systems), 20 ng/mL ACTIVIN A (Miltenyi Biotec), and 1.5 µM CHIR99021 (Axon Medchem). On day 3, cells were refreshed with BPEL supplemented with 5 µM XAV939 (Tocris Bioscience) and 50 ng/mL VEGF (R&D Systems). From day 6 onward, cells were refreshed every 3 days with BPEL medium supplemented with 50 ng/mL VEGF.

Cao X., Mircea M., Yakala G.K., van den Hil F.E., Brescia M., Mei H., Mummery C.L., Semrau S, & Orlova V.V. (2022). ETV2 Upregulation Marks the Specification of Early Cardiomyocytes and Endothelial Cells During Co-differentiation. Stem Cells, 41(2), 140-152.

+ Open protocol

+ Expand

Differentiation of iPSCs to Cardiomyocytes

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

Cell reprogramming into iPSC lines was previously described by Rovina et al. [15 (link)]. Briefly, iPSCs were seeded in Matrigel (Corning, 356,230) coated 6 well plates in StemFlex medium (ThermoFisher, A3349401) supplemented with RevitaCell (ThermoFisher, A2644501). On Day 0, medium was replaced with BPEL medium and cells were treated with 6 mM CHIR99021 (Bertin bioreagent, 13,122), 100 µg/ml Activin A (MiltenyiBiotec, 130–115-008), and 100 µg/ml BMP4 (R&D Systems, 5020-BP). On Day 3, cells were treated with 10 mM XAV939 (R&D Systems,3748), 100 µg/ml BMP4 and 2 mM Retinoic acid (Sigma Aldrich, R2625). Medium is changed on Day6 with BPEL + 30 ng/ml BMP4 + 1 µM Retinoic acid. After 3 days, cells were replated on fibronectin coated plates with BPEL medium supplemented with 20 mM SB 431542 (R&D Systems,161).

Soussi S., Savchenko L., Rovina D., Iacovoni J.S., Gottinger A., Vialettes M., Pioner J.M., Farini A., Mallia S., Rabino M., Pompilio G., Parini A., Lairez O., Gowran A, & Pizzinat N. (2023). IPSC derived cardiac fibroblasts of DMD patients show compromised actin microfilaments, metabolic shift and pro-fibrotic phenotype. Biology Direct, 18, 41.

+ Open protocol

+ Expand

Photocrosslinkable Hydrogel for Stem Cell Culture

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

PEG (Mn = 10 kg/mol), tin(II) 2-ethylhexanoate (Sn(Oct)₂), hydroquinone (HQ), triethylamine (TEA), methacrylic anhydride (MAAh), deuterated chloroform, 1,6-hexanediol, 2-hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophenone (Irgacure 2959), tris(hydroxymethyl)aminomethane (TRIS), Matrigel, verapamil hydrochloride (CAS RN: 52-53-9) and 3,4-dihydroxy-L-phenylalanine (L-DOPA) were purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands). Propylene carbonate (PC), ethanol absolute and calcium hydride were obtained from Merck Millipore (Darmstadt, Germany). Methanol, dichloromethane (DCM) and diethyl ether were purchased from VWR (Amsterdam, The Netherlands). Trimethylene carbonate (TMC) was obtained from Huizhou ForYou Medical Devices Co (Huizou, China). Essential 8 medium was purchased from Thermo Fisher (Naarden, The Netherlands), Wnt activator CHIR99021 (1.5 µmol/L) from Axon Medchem (Groningen, The Netherlands), Activin-A (20 ng/mL) from Miltenyi Biotec (Bergisch Gladbach, Germany) and BMP4 (20 ng/mL) and XAV939 (5 µmol/L) from R&D Systems (Abingdon, UK).

Allijn I., Ribeiro M., Poot A., Passier R, & Stamatialis D. (2020). Membranes for Modelling Cardiac Tissue Stiffness In Vitro Based on Poly(trimethylene carbonate) and Poly(ethylene glycol) Polymers. Membranes, 10(10), 274.

+ Open protocol

+ Expand

Directed Differentiation of hiPSCs into Cardiac Lineages

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

hiPSC line LUMC0099iCTRL#04 was maintained in mTESR1 medium (Stem Cell
Technologies, #5850) on growth factor reduced Matrigel, (Corning #356234) at
37°C/5% CO₂. For cardiomyocyte differentiation, cells were seeded at
a density of 10 × 10³ cells per cm². Differentiation was
induced when hiPSCs reached 80%–90% confluency. BPEL medium^{71 (link)}supplemented with 20 ng/mL Activin-A (Miltenyi Biotec #130-115-012), 20 ng/mL
BMP4 (R&D systems, #314-BP-101/CF) and 1.5 μM CHIR99021 (Axon Medchem,
#1386) was used to initiate differentiation to cardiac mesoderm. Three days
after initiation, medium was replaced with BPEL containing 5 μM XAV939
(Bioscience, #3748/10) for ventricular differentiation, in combination with BMS
753 (Tocris # 3505) for atrial differentiation or in combination with 2.5 ng/mL
BMP4 (R&D Systems 314-BP-010/CF), 5 μM SB431542 (Tocris #1614), 250 nM RA
and 250 nM PD173074 for sinoatrial nodal differentiation. Differentiation medium
was replaced with BPEL after 48–72 h, and cells were refreshed every 3 days
thereafter.

Li J., Wiesinger A., Fokkert L., Boukens B.J., Verkerk A.O., Christoffels V.M., Boink G.J, & Devalla H.D. (2022). Molecular and electrophysiological evaluation of human cardiomyocyte subtypes to facilitate generation of composite cardiac models. Journal of Tissue Engineering, 13, 20417314221127908.

+ Open protocol

+ Expand

Directed Cardiomyocyte Differentiation of hiPS Cells

Cited 1 time

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

Human iPS cells were differentiated into CMs using the established matrix sandwich method^{35 (link)} with modifications. Briefly, 6 days before initiating differentiation, hiPS cell colonies were passaged on human embryonic stem cell–qualified Matrigel-coated plates (0.05 mg/mL; BD Biosciences) using Gentle Cell Dissociation Buffer (Stemcell Technologies) and cultured as a monolayer in mTeSR1 with 1× Y-27632 ROCK inhibitor (Stemcell Technologies) in a normal oxygen atmosphere. When cells reached 80% confluence, cold mTeSR1 with Growth Factor Reduced Matrigel (0.033 mg/mL) was added to create an overlay of Matrigel. Differentiation was initiated 24 hours later (day 0) by culturing the cells in RPMI-1640 medium (Life Technologies) supplemented with B27 (without insulin; Life Technologies), 100 ng/mL activin A (Miltenyi Biotec), and 10 ng/mL FGF2 (Miltenyi Biotec) for 24 hours. On the next day, the medium was replaced by RPMI-1640 medium supplemented with B27 without insulin, 10 ng/mL BMP4 (Miltenyi Biotec), and 5 ng/mL FGF2 for 4 days. By day 5, cells were cultured in RPMI-1640 medium supplemented with B27 complete (Life Technologies) and 1% NEAA and changed every 2 to 3 days.

Gaignerie A., Lefort N., Rousselle M., Forest-Choquet V., Flippe L., Francois–Campion V., Girardeau A., Caillaud A., Chariau C., Francheteau Q., Derevier A., Chaubron F., Knöbel S., Gaborit N., Si-Tayeb K, & David L. (2018). Urine-derived cells provide a readily accessible cell type for feeder-free mRNA reprogramming. Scientific Reports, 8, 14363.

+ Open protocol

+ Expand

Differentiation of hiPSCs into ECs

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

hiPSCs were differentiated into hiPSC-ECs with a protocol by Orlova et al. [9 (link)] (S2A Fig). In short, hiPSCs were dissociated with TrypLE (Gibco) and seeded as single cells at 2x10⁴ cells/cm² on day -1 on vitronectin (Stem Cell Technologies, Vancouver, Canada) coated plates in E8 medium (Stem Cell Technologies) supplemented with ROCK inhibitor (ROCKi, Y-27632) (Stem Cell Technologies) (10uM). On day 0 cells were replated and cultured in BPEL medium (S1 Table) supplemented with VEGF (50ng/ml) (R&D Systems, Minneapolis, MN, United States), Activin A (25ng/ml) (Miltenyi, Bergisch Gladbach, Germany), BMP4 (30ng/ml) (R&D Systems) and CHIR99021 (1.5μM) (Tocris Bioscience, Bristol, United Kingdom). On day 3 medium was changed to BPEL supplemented with VEGF (25ng/ml) and SB431542 (10μM) (Tocris Bioscience), followed by a CD31 bead (Invitrogen) isolation step on day 10. Cells were then plated on 0.2% gelatin (Sigma-Aldrich, St. Louis, MO, United States) and cultured in expansion EC-SFM medium (Gibco) supplemented with VEGF (30ng/ml), FGF (20ng/ml) (Miltenyi), and PPP (1%) (Bio-Connect, Huissen, The Netherlands).

de Boer S., Laan S., Dirven R, & Eikenboom J. (2024). Approaches to induce the maturation process of human induced pluripotent stem cell derived-endothelial cells to generate a robust model. PLOS ONE, 19(2), e0297465.

+ Open protocol

+ Expand

Hepatocyte Differentiation Protocol

Cited 1 time

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

We followed the differentiation protocol published in^{19 (link)}. In brief, were plated in tissue-culture plates previously coated with Matrigel at 0.05 mg/ml for 1 h, and cultured in mTSeR or StemMACS^TM iPS-Brew XF. Once cells reached 70–80% confluence, differentiation was performed with RPMI 1640 (Life Technologies) and B27 (Life Technologies) containing Activin A (AA; Miltenyi Biotec), FGF2, BMP4 (Miltenyi Biotec) and HGF (Miltenyi Biotec) with the following sequence: AA/BMP4/FGF2 (2 days); AA (3 days), BMP4/FGF2 (5 days) and HGF (5 days). Cells were then incubated with Hepatocyte Culture Medium (Lonza) supplemented with oncostatin M (Miltenyi Biotec) for 5 days.

+ Open protocol

+ Expand

Optimizing hESC Pluripotency and Cardiac Differentiation

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

For the first 24 h after hESC seeding (day -1 to day 0), the E8 medium in the chambers was fully exchanged in programmatically set intervals. To test the effect of different medium exchange intervals on hESC pluripotency and proliferation, intervals between 1 and 8 h were chosen for different chambers. For cardiac mesoderm differentiation experiments, the exchange interval was set to 2 h for all chambers. On day 0, the cells were either fixed (for pluripotency staining) or the chambers were all flushed with BPEL medium and subsequently growth factor-supplemented (20 ng/mL Activin A (Miltenyi Biotech), 20 ng/mL BMP-4 (R&D Systems), and 1.5 µM CHIR-99021 (Sigma Aldrich)) BPEL medium. The program was reset to exchange the medium at new intervals (between 1 and 5 h). Every medium exchange had a duration of 7 s per chamber and the pressure for the flow was set between 35 and 40 mbar.

Vollertsen A.R., Den S.A., Schwach V., van den Berg A., Passier R., van der Meer A.D, & Odijk M. (2021). Highly parallelized human embryonic stem cell differentiation to cardiac mesoderm in nanoliter chambers on a microfluidic chip. Biomedical Microdevices, 23(2), 30.

+ Open protocol

+ Expand

Mesoderm Differentiation of MESP1-NKX2.5 hESCs

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

A previously generated MESP1^mCherry/w-NKX2.5^eGFP/w human embryonic stem cell (hESC) dual reporter line (Den Hartogh et al. 2015 (link)) was maintained in standard wells plates coated with vitronectin (Thermo Fisher) and cultured in Essential 8 (E8) medium (Thermo Fisher) supplemented with 1% penicillin/streptomycin (Thermo Fisher). HESCs were passaged twice a week using EDTA (Thermo fisher).
Control experiments for cell differentiation were performed in 6-well plates in parallel to on-chip differentiation. On day -1, hESCs were seeded at 125 × 10³ cells/mL in E8 medium supplemented with 1% penicillin/streptomycin on a Matrigel-GFR (VWR) coated 6-well plate. On day 0, the cells were washed with in-house made BPEL (BSA, Polyvinyl alcohol, Essential Lipids) medium (Ng et al. 2008 (link)), before adding growth factor-supplemented medium (20 ng/mL Activin A (Miltenyi Biotech), 20 ng/mL BMP-4 (R&D Systems), and 1.5 µM CHIR-99021 (Sigma Aldrich)) in BPEL medium at 3 mL per well. The differentiation timeline is shown in Fig. 6. All cell cultures were maintained at 5% CO₂, 37 °C, and atmospheric O₂ levels.
Fig. 6

A schematic overview of the differentiation protocol towards mesodermal lineage. MESP1^mCherry expression is indicated in red around day 3

+ 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!