Vitronectin Cytocaf®
Pack size: 1 mL, 1 mg/mL
Price: £48
Available from TCS Biosciences Ltd.
Features checklist
- Provides adhesive surface: Ideal for cells such as iPSCs
- Easy to handle: No need to keep cool, can be used at room temperature and above without denaturing or losing activity
- Simple 1 hour protocol for coating plates
- Free from animal material
Product description
Cytocaf® proteins are based on the bacterial protein Caf1, which associates into long, flexible and bioinert polymers. Bioactivity can be imparted by genetically encoding bioactive motifs into the protein’s sequence. Cytocaf polymers are highly stable, modular and flexible.
Vitronectin Cytocaf® mimics the action of the extracellular matrix protein vitronectin and is adhesive for a range of cell types such as human mesenchymal stromal cells (MSCs) and induced pluripotent stem cells (iPSCs).
Cells grown on Vitronectin Cytocaf® display adherence and morphologies similar to those seen on recombinant vitronectin and Geltrex™
Vitronectin/Integrin Interaction
Vitronectin Cytocaf®/Integrin Interaction
References
- Dura, G., Crespo-Cuadrado, M., Waller, H., Peters, D. T., Ferreira-Duarte, A., Lakey, J. H. & Fulton, D. A. (2022). Exploiting Meltable Protein Hydrogels to Encapsulate and Culture Cells in 3D. Macromolecular Bioscience 22, 2200134
- Dura, G., Crespo-Cuadrado, M., Waller, H., Peters, D. T., Ferreira, A. M., Lakey, J. H. & Fulton, D. A. (2021). Hydrogels of engineered bacterial fimbriae can finely tune 2D human cell culture. Biomaterials Science 9, 2542-2552.
- Dura, G., Peters, D. T., Waller, H., Yemm, A. I., Perkins, N. D., Ferreira, A. M., Crespo-Cuadrado, M., Lakey, J. H. & Fulton, D. A. (2020). A Thermally Reformable Protein Polymer. Chem 6, 3132-3151.
- Roque, A. I., Soliakov, A., Birch, M. A., Philips, S. R., Shah, D. S. & Lakey, J. H. (2014). Reversible non-stick behaviour of a bacterial protein polymer provides a tuneable molecular mimic for cell and tissue engineering. Adv Mater 26, 2704-2709, 2616.
Data & specifications
| Test | Result |
|---|---|
| Bacterial growth | Negative |
| Fungal growth | Negative |
| Particulate examination | Negative |
| Endotoxin level | < 1 EU/µg |
Figure 1: SDS-PAGE showing purified Vitronectin-Cytocaf, heated (monomer) and unheated (polymer).
Figure 2: (A) iPSCs derived from human dermal fibroblasts were cultured on non-TC plastic culture dishes coated with 2µg/cm² Vitronectin-Cytocaf. Images were taken at 10X magnification, 48 hours after seeding, by bright field microscopy. (B) Non-TC plastic was either left uncoated, coated with unmodified Cytocaf or Vitronectin-Cytocaf batch 8 prior to seeding hDfa iPSCs in E8 Flex media supplemented with 10 µM ROCK inhibitor. After 24 hours media was exchanged to remove the inhibitor. A further 24 hours later, cell viability was measured using the Presto Blue cell viability assay, as per manufacturer’s specifications.
Figure 3: iPSCs were seeded onto non-TC plastic coated with 2 µg/mL of Vitronectin-Cytocaf. Cells were grown in E8 media and subcultured using ReleasR (Stem Cell Technologies) as a dissociation reagent. At passage 1, 3 and 5 cells were harvested for RNA analysis. RNA was harvested, reverse transcribed to cDNA and analysed using quantitative real-time PCR. OCT4, SOX2, and NANOG are well-known markers of iPSCs, and CD44 was used as the negative marker. The graph displays the average expression of positive and negative markers across three independent batch tests of Vitronectin Cytocaf. Error bars represent SEM.
Figure 4: Heat map of gene expression fold changes from TaqMan hiPSC scorecard assay with mesoderm, ectoderm, and endoderm gene markers highlighted. Color range illustrates reduced increasing expression (red), gene identifier shown below. Boxplots correspond to center quartiles for all genes in each linage set, the mean is marked by a black bar and whiskers show total range of values. Significance between sets determined by one way ANOVA coupled with Šídák’s correction. Dotted lines plot baseline reference levels from supplier (<-1: downregulation, >+1: upregulation) plotted as Log2 of fold change. ****p < 0.0001. Adapted from Creigh et al., 2024, bioRxiv 2024.04.29.591606