Sphere formation from corneal keratocytes and phenotype specific markers

Area of Science:

• Ophthalmology
• Cell Biology
• Tissue Engineering

Background:

• Keratocytes are crucial mesenchymal cells for corneal stroma maintenance.
• Phenotypic plasticity of keratocytes into fibroblasts/myofibroblasts impacts corneal transparency.
• Understanding keratocyte behavior is vital for corneal tissue engineering.

Purpose of the Study:

• Investigate sphere-forming capabilities of human and bovine keratocytes.
• Identify distinguishing markers for keratocyte and fibroblast phenotypes.
• Optimize culture conditions for preserving keratocyte phenotype.

Main Methods:

• Isolated human and bovine keratocytes.
• Cultured cells in serum-free and serum-containing media.
• Analyzed sphere formation under varying attachment conditions.
• Assessed expression of vimentin, keratocan, BMP3, and cadherin 5.

Main Results:

• Keratocyte morphology and phenotype were modulated by serum availability.
• Bovine keratocytes formed spheres in adherent and low-attachment conditions; human keratocytes only in low-attachment.
• Vimentin was expressed in both keratocytes and fibroblasts.
• Keratocan was found in early bovine fibroblasts.
• BMP3 was detected in keratocytes and spheres; cadherin 5 in keratocytes only.

Conclusions:

• Keratocyte sphere formation varies between species and culture conditions.
• BMP3 and cadherin 5 show potential as specific keratocyte markers.
• These findings aid in developing strategies for corneal tissue regeneration.