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Several Stem Cell Core Facility Directors Explain Why They Choose R&D Systems Cytokines

Directors at several different stem cell core facilities describe their experiences using R&D Systems cytokines and growth factors. The speakers on the video include Sunita D’Souza, Director of the Center for Modern Pediatric Diseases at St. Jude’s Medical Center, Deborah French, Director of the Human Pluripotent Stem Cell Core at the Children’s Hospital of Philadelphia, Barbara Corneo, Director of the Stem Cell Core Facility at Columbia University Medical School, and Wenli Yang, Director of the iPSC Core Facility at the University of Pennsylvania. Learn why they use R&D Systems cytokines for their research. 

Pluripotent Stem Cells

Cytokines for ESC and iPSC Expansion - Products by Molecule

Activin A * BMP-4 * FGF basic/FGF2 * Nodal TGF-beta 1 * Wnt-3a *

Cytokines for Ectoderm Lineage Differentiation - Products by Molecule

BMP-4 * EGF * FGF basic/FGF-2 * Noggin *

Cytokines for Mesoderm Lineage Differentiation - Products by Molecule

Activin A * BMP-2 * BMP-4 * Dkk-1
FGF basic/FGF2 * FGF-3 FGF-10 FGF-17
Flt-3 Ligand * IL-3 * IL-6 * IL-11
Noggin * PDGF-BB * SCF/c-kit Ligand * TGF-beta 1 *
Thrombopoietin/TPO * VEGF Wnt-5a  

Cytokines for Endoderm Lineage Differentiation - Products by Molecule

Activin A * BMP-4 * EGF * FGF basic/FGF-2 *
FGF-4 FGF-7/KGF * FGF-10 FGF-18
HGF * IGF-I/IGF-1 * Noggin * PDGF-BB *
Sonic Hedgehog/Shh * VEGF Wnt-3a *  

*GMP-grade Proteins are available for these molecules.

ExCellerate™ iPSC Expansion Medium

New ExCellerate iPSC Expansion Medium

Supports robust expansion and maintenance of pluripotent stem cell culture for enhanced consistency and reproducibility.

  • Animal component-free
  • No growth factor supplementation required
  • Stable cell integrity over long term culture

Assessment of the Bioactivity of R&D Systems Recombinant Human FGF2 and Wnt-3a

A. Assessment of the bioactivity of R&D Systems Recombinant Human FGF basic and compared with a top competitor’s FGF basic B. (Data image from Catalog # 5036-WN – change the y-axis label to Alkaline Phosphatase Production (Mean O.D.)

R&D Systems Recombinant Human FGF basic/FGF2 (146 aa) and Recombinant Human Wnt-3a Display Higher Activity than the Same Proteins Provided by Leading Competitors.

(A) The bioactivity of R&D Systems Recombinant Human FGF basic/FGF2 (146 aa) (Catalog # 233-FB; orange line) or recombinant human FGF basic/FGF2 (146 aa) from a leading competitor (green line) was assessed by measuring the ability of the proteins to stimulate proliferation of the NR6R-3T3 mouse fibroblast cell line. The bioactivity of R&D Systems Recombinant Human FGF basic/FGF2 (146 aa) was approximately 3-fold greater than the top competitor’s FGF basic (146 aa). (B) The bioactivity of R&D Systems Recombinant Human Wnt-3a (Catalog # 5036-WN; orange line) or recombinant human Wnt-3a from another company (green line) was assessed by measuring the ability of the proteins to induce alkaline phosphatase production in the MC3T3-E1 mouse preosteoblast cell line. The ED50 for this effect for R&D Systems Recombinant Human Wnt-3a was 1.7-fold better, with more than twice the maximum response compared to the competitor's Wnt-3a.

Differentiation of hESCs into Endoderm Induced by Recombinant Human Activin A

A. Claudin-6 and Sox17 immunostaining of human embryonic stem cells differentiated into endoderm in Activin A-supplemented media B. Claudin-6 and Sox17 immunostaining of human embryonic stem cells cultured in media without Activin A

Differentiation of Human Embryonic Stem Cells in Media Supplemented with R&D Systems Recombinant Human Activin A.

BG01V human embryonic stem cells were (A) cultured in media without any protein added or (B) differentiated into endoderm using media supplemented with Recombinant Human Activin A (R&D Systems, Catalog # 338-AC). Differentiation into endoderm was confirmed by positive staining for Claudin-6 (red) and Sox17 (green) using the Mouse Anti-Human Claudin-6 Monoclonal Antibody (R&D Systems, Catalog # MAB3656) and the Goat Anti-Human Sox17 Polyclonal Antibody (R&D Systems, Catalog # AF1924), respectively. BG01V human embryonic stem cells are licensed from ViaCyte, Inc. 

Neural Stem Cells

Cytokines for Neural Stem Cells - Products by Molecule

BDNF BMP-2 * BMP-4 * CNTF EGF * FGF basic/FGF2 *
FGF-4 FGF-8a FGF-8b FGF-10 GDNF * IGF-I/IGF-1 *
beta-NGF Noggin * NT-4 PDGF-BB * Sonic Hedgehog/Shh * VEGF

*GMP-grade Proteins are available for these molecules.

Analysis of the Bioactivity and Consistency of R&D Systems Recombinant Human Noggin

ASOX1, Otx2 staining of human embryonic stem cells differentiated to neuroectoderm with FGF basic and Recombinant Human Noggin B Quantification of SOX1 clusters in human embryonic stem cells incubated with R&D Systems Noggin or Noggin from two competitors

R&D Systems Recombinant Human Noggin Displays Minimal Lot-to-Lot Variability and Higher Activity than Leading Competitors’ Noggin Proteins.

BG01V human embryonic stem cells were cultured in Mouse Embryonic Fibroblast Conditioned Media supplemented with FGF basic (5 ng/mL). Stem cells were driven into early cells of the neuroectoderm using a 3-day incubation in Recombinant Human Noggin (25 ug/mL) from either R&D Systems (Lot 1, Lot 2; Catalog # 6057-NG) or from two different competitors (Competitor 1, Competitor 2). Control cells were incubated in media without Noggin (No Noggin). The cells were stained for the early ectoderm marker, Otx2, and the neuroectoderm marker, SOX1. (A) Representative images of SOX1 (green), Otx2 (red), and DAPI (blue) staining in embryonic stem cells differentiated with Noggin from R&D Systems or Noggin from Competitor 2. (B) SOX1+ clusters were quantified under each of the indicated culture conditions. Cells treated with R&D Systems Noggin showed an increase in SOX1+ cells compared to both untreated and competitor-treated cells. R&D Systems Noggin showed consistent differentiation across the lots tested. BG01V human embryonic stem cells were licensed from ViaCyte, Inc. (C) The bioactivity of R&D Systems Recombinant Human Noggin (Catalog # 6057-NG; orange line) or recombinant human noggin from a top competitor (green line) was determined by assessing the ability of the proteins to inhibit alkaline phosphatase production induced by 50 ng/mLRecombinant Human BMP-4 (R&D Systems, Catalog # 314-BP) in the ATDC5 mouse chondrogenic cell line. In the presence of 50 ng/mL Recombinant Human BMP-4, the ED50 for this effect for R&D Systems Recombinant Human Noggin was approximately 30-fold greater than the top competitor’s Noggin protein. 

Mesenchymal Stem Cells

Cytokines for Mesenchymal Stem Cells - Products by Molecule

BMP-2 * BMP-4 * BMP-6 EGF * FGF basic/FGF2 * FGF-4
GDF-5/BMP-14 HB-EGF IGF-I/IGF-1 * LR3 IGF-I/IGF-1 * IL-6 * PDGF-BB *
Sonic Hedgehog/Shh * TGF-beta 1 * TGF-beta 2 TGF-beta 3 Wnt-5b Wnt-10b

*GMP-grade Proteins are available for these molecules.

Analysis of the Bioactivities of R&D Systems Recombinant Human Shh Proteins

A. Assessment of the activity of HEK293-purified High Activity Recombinant Human Shh compared with E.coli-purified Shh proteins B. LC/ESI-MS analysis of HEK293-purified High Activity Recombinant Human Shh identifies cholesterol and fatty acid modification

R&D Systems Recombinant Human Sonic Hedgehog/Shh Purified from HEK293 Cells and Containing the Correct Post-Translational Modifications Displays Higher Activity than E. coli-purified Recombinant Human Shh Proteins.

(A) The bioactivity of R&D Systems High Activity Recombinant Human Shh (Catalog # 8908-SH; green line), purified from HEK293 cells and containing the correct post-translational modifications (cholesterol and fatty acids), was compared to the bioactivity of E. coli-purified Recombinant Human Shh (C24II) N-terminus (R&D Systems, Catalog # 1845-SH; red line), and E.coli-purified Recombinant Human Shh, N-terminus (R&D Systems, Catalog # 1314-SH; blue line) by measuring the ability of the three proteins to induce alkaline phosphatase production by mesenchymal stem cells. The HEK293-purified High Activity Recombinant Human Shh protein was found to be over 14-fold more active than the E.coli-purified Recombinant Human Shh (C24II) N-Terminus protein, and over 250-fold more active than E. coli-purified Recombinant Human Shh-N protein. (B) LC/ESI-MS analysis of High Activity Recombinant Human Shh (R&D Systems, Catalog # 8908-SH) shows major peaks at 20119.3, 20145.2, and 20171.6 Da, suggesting that Recombinant Human Shh molecules are dual-modified with cholesterol at the C-terminus, and fatty acids (lauric acid, myristic acid, and palmitic acid) at the N-terminus. The minor peaks at 19776 Da corresponds to Recombinant Human Shh with only the fatty acid modification. 

Hematopoietic Stem Cells

Cytokines for Hematopoietic Stem Cells - Products by Molecule

BMP-4 * Erythropoietin Flt-3 Ligand * G-CSF
GM-CSF * IFN-gamma * IL-2 * IL-3 *
IL-4 * IL-6 * IL-7 * IL-10
IL-11 IL-15 * IL-21 * M-CSF *
SCF/c-kit Ligand * Thrombopoietin * VEGF Wnt-3a *

*GMP-grade Proteins are available for these molecules.

Assessment of the Bioactivity of R&D Systems Recombinant Human Thrombopoietin

Assessment of the bioactivity of R&D Systems Recombinant Human Tpo and compared with two leading competitors’ Tpo proteins

R&D Systems Recombinant Human Thrombopoietin/Tpo Displays Higher Activity than Leading Competitors’ Thrombopoietin Proteins.

The bioactivity of R&D Systems Recombinant Human Thrombopoietin (Catalog # 288-TPE; orange line) or recombinant human Thrombopoietin/Tpo from two different competitors (green and purple lines) was assessed by measuring the ability of the proteins to stimulate proliferation of the MO7e human megakaryocytic leukemic cell line. The ED50 for this effect for R&D Systems Recombinant Human Thrombopoietin is 0.05-0.5 ng/mL, which is over 2-fold more active than the two competitors’ proteins.

Consistency Testing of R&D Systems GMP-grade Recombinant Human Flt-3 L and SCF

A. Assessment of the lot-to-lot consistency of GMP-grade Recombinant Human Flt-3 Ligand using a cell proliferation assay B. Assessment of the lot-to-lot consistency of GMP-grade Recombinant Human SCF using a cell proliferation assay

R&D Systems GMP-grade Recombinant Human Flt-3 Ligand and GMP-grade Recombinant Human SCF Display High Lot-to-Lot Consistency.

A) Three independent lots of GMP-grade Recombinant Human Flt-3 Ligand (R&D Systems, Catalog # 308E-GMP) were tested for their ability to stimulate proliferation of the BaF3 mouse pro-B cell line transfected with mouse Flt-3. The ED50 for this effect is 0.2-1 ng/mL. (B) Three independent lots of GMP-grade Recombinant Human SCF (R&D Systems, Catalog # 255B-GMP) were tested for their ability to stimulate proliferation of the TF-1 human erythroleukemic cell line. The ED50 for this effect is 1-5 ng/mL. Each trace shown on the graphs represents data obtained from GMP-grade Recombinant Human Flt-3 Ligand or GMP-grade Recombinant Human SCF from a different manufacturing run to show the lot-to-lot consistency of the proteins.

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Embryonic stem (ES) cells and induced pluripotent stem (iPS) cells are self-renewing progenitors that have the capacity to differentiate into cells of the ectoderm, mesoderm, and endoderm. Explore this poster to learn about the markers that can be used to identify different stem cell lineages.

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Download this guide to learn about the products that we offer for isolating, differentiating, verifying, and investigating embryonic and induced pluripotent stem cells. From specialized media and growth factors to small molecules and antibody kits, our reagents are designed to reduce experimental variability and improve the consistency of your data.

Neural Stem Cells Product Guide

Learn about our portfolio of reagents designed to simplify the culture, expansion, and differentiation of neural stem cells. We offer a wide selection of serum-free neural media supplements, growth factors, and small molecules, along with neural lineage differentiation and functional identification kits to optimize your neural stem cell-related research. 

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Organoid Culture Handbook

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