Activin a

Manufactured by Sino Biological

Activin A is a protein that belongs to the transforming growth factor-beta (TGF-β) superfamily. It plays a key role in the regulation of various biological processes, such as cell growth, differentiation, and homeostasis. Activin A is involved in the activation of the SMAD signaling pathway and can modulate the expression of target genes.

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Lab products found in correlation

Bone morphogenetic protein 4 (bmp4), by Thermo Fisher Scientific (4 mentions) Basic fibroblast growth factor (bfgf), by Sino Biological (4 mentions) Stem cell factor (scf), by Thermo Fisher Scientific (3 mentions) Chir99021, by Selleck Chemicals (3 mentions) Vascular endothelial growth factor (vegf), by Sino Biological (3 mentions) Basic fibroblast growth factor (bfgf), by Selleck Chemicals (3 mentions) Ly294002, by Selleck Chemicals (3 mentions) Sb431542, by Selleck Chemicals (3 mentions) Matrigel, by Corning (2 mentions) Accutase, by Merck Group (2 mentions) A83 01, by Selleck Chemicals (2 mentions) Thiazovivin, by Selleck Chemicals (2 mentions) Thrombopoietin, by Sino Biological (2 mentions) Iwr 1 endo, by Selleck Chemicals (2 mentions) Flt3l, by Thermo Fisher Scientific (2 mentions) Vascular endothelial growth factor (vegf), by Selleck Chemicals (2 mentions) Interleukin 3 (il 3), by Sino Biological (2 mentions) Interleukin 6 (il 6), by Sino Biological (2 mentions) Insulin transferrin selenium, by Thermo Fisher Scientific (1 mentions) Vitamin c, by Merck Group (1 mentions)

5 protocols using activin a

Directed Hematopoietic Differentiation of hPSCs

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Before haematopoietic differentiation, hPSCs were dissociated by Accutase (Sigma) and plated on growth factor‐reduced Matrigel (Corning)‐coated plates with thiazovivin (0.1 μM, Selleck). Firstly, at day 0, 40 ng/ml of BMP4 (Peprotech), 30 ng/ml of ACTIVINA (Sino Biological Inc.), 20 ng/ml of bFGF (Sino Biological Inc.), 6 μM CHIR99021 (Selleck) and 10 μM LY294002 (Selleck) were added to the basic medium (BM, mimics of the CustommTeSR1) of Dulbecco's‐modified Eagle's medium/F‐12 (GIBCO) supplemented with 1% insulin–transferrin–selenium (GIBCO), 70 μg/ml of vitamin C (Sigma). Second, 30 ng/ml of BMP, 1 μM A8301 (Selleck) and 2 μM IWR‐1‐endo (Selleck) were added to the BM on day 1. Then, on days 2–4 of differentiation, 40 ng/ml of vascular endothelial growth factor (Sino Biological Inc.) and 50 ng/ml of bFGF were added to the BM. Finally, 40 ng/ml of vascular endothelial growth factor, 50 ng/ml of bFGF, 10 μM SB431542 (Selleck), 10 ng/ml of stem cell factor (Peprotech), 50 ng/ml of thrombopoietin (Sino Biological Inc.), 10 ng/ml ofinterleukin 3 (Sino Biological Inc.) and 50 ng/ml of interleukin 6 (Sino Biological Inc.) were added in the BM at days 4–6 of differentiation and further haematopoietic commitment and maturation.

Kang B., Zhang T., Huang K., Wang T., Li Y., Mai Y., Li J., Dang S., Zhang Z., Huang W., Wang J., Gao M., Wang Y, & Pan G. (2022). GFI1 regulates chromatin state essential in human endothelial‐to‐haematopoietic transition. Cell Proliferation, 55(5), e13244.

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Hematopoietic Differentiation of H1 hESCs

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Hematopoietic differentiation was performed as previously described (Zhu et al., 2020 (link)). Briefly, H1 hESCs were plated onto Growth Factor Reduced Matrigel (1:200 dilution; BD Biosciences)-coated plates at a proper initial density. The hematopoietic differentiation medium was changed every day, supplemented with corresponding cytokines or inhibitors, and floated hESCs-HSPCs were collected from D8 for further assays. The cytokines or inhibitors added each day were as follows: D0–D1, 40 ng/mL BMP4 (PeproTech), 30 ng/mL Activin A (Sino Biological), 20 ng/mL bFGF (Sino Biological), 6 μM CHIR99021 (Selleck), and 10 μM LY294002 (Selleck); D1–D2, 30 ng/mL BMP4, 1 μM A8301 (Selleck), and 2 μM IWR-1-endo (Selleck); D2–D4, 40 ng/mL vascular endothelial growth factor (VEGF) (Sino Biological) and 50 ng/mL bFGF; D4 and later, 40 ng/mL VEGF, 50 ng/mL bFGF, 10 μM SB431542 (Selleck), 10 ng/mL SCF (PeproTech), 50 ng/mL TPO (Sino Biological), 10 ng/mL IL3 (Sino Biological), 50 ng/mL IL6 (Sino Biological), and 50 ng/mL Flt3L (PeproTech).

Gu J., Zhu Y., Lin H., Huang Y., Zhang Y., Xing Q., Kang B., Zhang Z., Wang M., Zhou T., Mai Y., Chen Q., Li F., Hu X., Wang S., Peng J., Guo X., Long B., Wang J., Gao M., Shan Y., Cui Y, & Pan G. (2024). Autophagy is essential for human myelopoiesis. Stem Cell Reports, 19(2), 196-210.

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Hematopoietic Differentiation of hESCs

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To initiate hematopoietic differentiation^{42 (link)}, the hESCs were passaged with dispase (2 mg mL⁻¹) onto matrigel-coated 12-well plates. These cells were cultured in E6 medium^{46 (link)} plus ACTIVIN A and BMP4 (DMEM/F12 (Hyclone), 64 mg L⁻¹ Lascorbic acid (Sigma), NaCl (Sigma, adjusting the osmolarity to 340 mOsm), ITS –G (Gibco, 100×) and 50 ng mL⁻¹ ACTIVIN A (Sino biological, 10429-HNAH-50), 50 ng mL⁻¹ BMP4 (Peprotech, 120-05ET)) for 2 days. And then cells were cultured in E6 medium plus 40 ng mL⁻¹ VEGF (Sino biological, 10008-HNAB-50) and 50 ng mL⁻¹ bFGF (Sino biological, 10014-HNAE-50) for next two days. For next 3 days, these cells were cultured in E6 medium plus 40 ng mL⁻¹ VEGF, 50 ng mL⁻¹ bFGF, 10 μM SB431542 (Selleck). After 7 days, these cells were collected for extracting total RNA and FACS analysis for CD34 expression. CD34 antibody conjugated with PerCP-Cy5.5 were used according to the manufacturer’s recommendations.

Shan Y., Liang Z., Xing Q., Zhang T., Wang B., Tian S., Huang W., Zhang Y., Yao J., Zhu Y., Huang K., Liu Y., Wang X., Chen Q., Zhang J., Shang B., Li S., Shi X., Liao B., Zhang C., Lai K., Zhong X., Shu X., Wang J., Yao H., Chen J., Pei D, & Pan G. (2017). PRC2 specifies ectoderm lineages and maintains pluripotency in primed but not naïve ESCs. Nature Communications, 8, 672.

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Stepwise Differentiation of hPSCs to Macrophages

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hPSCs were dissociated by Accutase (Sigma) and plated on growth factor-reduced Matrigel (Corning)-coated plates with thiazovivin (0.1 μM, Selleck). hPSCs were induced for stepwise blood differentiation in basal medium supplemented with cytokines and inhibitors.²³ (link) Briefly, hPSCs were treated with DMEM/F12 (Gibco) medium containing 40 ng/mL BMP4 (Peprotech), 30 ng/mL ACTIVIN A (Sino Biological), 20 ng/mL basic fibroblast growth factor (bFGF) (Sino Biological), 6 μM CHIR99021 (Selleck), and 10 μM LY294002 (Selleck) for 1–2 days. Then, we switched to medium containing 40 ng/mL vascular endothelial growth factor (Sino Biological) and 50 ng/mL bFGF for 2 or 3 days. HSPCs were induced by medium containing 10 ng/mL SCF (Peprotech), 50 ng/mL thrombopoietin (Sino Biological), 10 ng/mL IL-3 (Sino Biological), 50 ng/mL IL-6 (Sino Biological), and 50 ng/mL FMS-like tyrosine kinase 3 ligand (FLT3L) (Peprotech). Floating iHSPCs were collected and plated in myeloid differentiation medium (StemPro +50 ng/mL FLT3L + 50 ng/mL M-CSF + 25 ng/mL GM-CSF) for 12 days and switched to macrophage maturation medium (RPMI-1640 + 10% FBS + 50 ng/mL M-CSF) for 7 days. All of the recombinant human cytokines were purchased from Sino Biological.

Kang B., Xing Q., Huang Y., Lin H., Peng J., Zhang Z., Wang M., Guo X., Hu X., Wang S., Wang J., Gao M., Zhu Y, & Pan G. (2024). Large-scale generation of IL-12 secreting macrophages from human pluripotent stem cells for cancer therapy. Molecular Therapy. Methods & Clinical Development, 32(1), 101204.

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Inducing Intestinal Stem Cell-like Cells from BM-MSCs

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BM-MSCs of passage 3 were seeded onto six-well plates. After adherence, the medium was changed to DMEM+1% FBS. Different concentrations of Activin A (0, 1, 5, 10, 20, and 100 ng/mL) were added to the medium. Activin A was purchased from Sino Biological. After 5 days, the expression of Foxa2 and Sox17 was measured by qRT-PCR and western blot. Then the optimal concentration of Activin A was used in the following experiment. For ISC-like cell differentiation, DMEM/F12 (GIBCO)+2% FBS supplemented with 250 ng/mL FGF2 (R&D Systems) was applied to the cells for 4 days. Lgr5 and Musashi-1, two ISC markers, were evaluated at the end of the protocol. To further observe whether the induced ISC-like cells could differentiate into intestinal epithelial cells, we cultured cells in DMEM/F12 containing 2% FBS and 20 ng/mL EGF (R&D systems) for 16 days. The medium was changed every 2 days.

Ye L., Sun L.X., Wu M.H., Wang J., Ding X., Shi H., Lu S.L., Wu L., Wei J., Li L, & Wang Y.F. (2018). A Simple System for Differentiation of Functional Intestinal Stem Cell-like Cells from Bone Marrow Mesenchymal Stem Cells. Molecular Therapy. Nucleic Acids, 13, 110-120.

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