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LamininB4-Cytocaf

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LamininB4-Cytocaf mimics the action of the extracellular matrix protein Laminin (Beta 4 Subunit) through a genetically encoded bioactive peptide that can bind to integrin receptors. This makes it adhesive for a range of cell types including both human and animal iPSCS.

Key Features:

Recombinant protein polymer that mimics the action of Laminin Beta 4 for adhesive cell culture:

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.

References

1. Adam R. Creigh, Helen Waller, Jeremy H. Lakey, Zofia M. Chrzanowska-Lightowlers, Robert N. Lightowlers, Daniel T. Peters Engineering bacterial protein polymers to support human pluripotent stem cell growth and differentiation in culture bioTxiv 2024.04.29.591606; doi: https://doi.org/10.1101/2024.04.29.591606

Data & Specifications

Test	Result
Bacterial growth	Negative
Fungal growth	Negative
Particulate examination	Clear
Mycoplasma	Negative
Host Cell DNA	Undetectable

Human iPSCs were seeded onto LamininB4-Cytocaf coated non tissue culture treated 3.5 cm plates and grown for three days in E8 flex media, before imaging with a light microscope at 10X magnification.

Human iPSCs were seeded at 0.5 x 104 cells per well in wells of a non-tissue culture treated 96-well plastic plate that were either (i) uncoated, (ii) coated with unmodified Cytocaf or (iii) coated with different batches of LamininB4-Cytocaf, in E8 flex media supplemented with CloneR2. After 24 h the media was exchanged for fresh E8 flex media and cells were then grown for a further 24 hours. Cell viability was then measured using a PrestoBlue assay according to the manufacturer’s instructions. Error bars represent SEM from three technical replicates.

After 5 passages of human iPSCs on LamininB4-Cytocaf coated plates, cells were seeded at a split ratio of 1 in 5 into LowAdhere-Cytocaf (which is non-adhesive for iPSCs) coated 6 cm non-TC treated dish in E8 flex media supplemented with 4 mg/mL polyvinyl alcohol (PVA) and 10 µM ROCKi. After 24 hours the media was changed to E6 media with 4 mg/mL PVA. The same media exchange was performed every 2-3 days for 7 days before embryoid bodies were harvested for qPCR analysis.

An ideal growth substrate for adherent cells.