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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.