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GelEnhance-Cytocaf

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A simple additive for 3D materials that enhances cell motility and viability.

GelEnhance-Cytocaf is a formulation of the unmodified Cytocaf (Caf1) material. The native function of the Cytocaf protein is to prevent the adherence of mammalian cells to bacteria, and so it has evolved over millennia to be perfectly engineered for this purpose.

By adapting the Cytocaf protein, we are able to exploit this functionality to provide a simple additive for 3D materials, such as basement membrane extract (BME) hydrogels, that enhances cell motility and viability.

Key Features:

Recombinant protein polymer for the enhancement of basement membrane extract gels:

Improves cell motility
Increases the number of vial cells
Improves vascular network formation ~3x
Simply mixes in with existing hydrogels

HEK293 cells were seeded at 1 x 10⁴ cells per well encapsulated in 1% w/v alginate or 1% w/v alginate supplemented with 0.2% w/v GelEnahnce-Cytocaf. Per well, 100 µL of the cell and hydrogel mixtures were added to 75 µL of 25 mM calcium chloride and incubated at 37°C and 5% CO for 1 hour to allow gelation. Following that, 100 µL of cell culture media was added per well. Two media formulations were used in this experiment comparing DMEM supplemented with 10% FBS (normal serum conditions) and DMEM supplemented with 2% FBS (low serum conditions). At regular intervals, a PrestoBlue assay was performed with the fluorescence reading correlating to the relative number of viable cells per well. Error bars represent SEM from three replicates.

Human umbilical cord derived endothelial cells (Angicyte) were seeded into Matrigel either neat or supplemented with GelEnhance-Cytocaf, then grown for three days in EGM-2 media supplemented with 10% FBS and imaged with a light microscope at 10x magnification.

Matrigel Only

Matrigel + 10 µg/mL GelEnhance-Cytocaf

Matrigel + 20 µg/mL GelEnhance-Cytocaf

2.5 x 10⁴ HCT116 cells were encapsulated per droplet of 1% w/v alginate or 1% alginate/0.2% w/v GelEnhance-Cytocaf. Droplets were formed by mixing 7.5 µl of cell/gel mixture with 5 µl 50 mM CaCl₂. Droplets were incubated at 37°C for 45 minutes. DMEM supplemented with 10% FBS was added to each well and images taken at the point of seeding and following four days of growth. ImageJ analysis was performed to calculate the % migrated across the four days.