Establishment and characterization of Caspian horse fibroblast cell bank in Iran

Area of Science:

• Equine genetics
• Conservation biology
• Cell culture technology

Background:

• The Caspian horse, a rare breed originating from Iran, faces a significant risk of extinction.
• Urgent conservation strategies are needed to preserve this unique equine genetic resource.

Purpose of the Study:

• To establish and characterize fibroblast cell lines from Caspian horses for conservation and research.
• To create a cryopreserved cell bank for the long-term preservation of Caspian horse genetic material.

Main Methods:

• Primary explant technique was used to establish 51 fibroblast cell lines from ear marginal tissue of 60 Caspian horses.
• Cell line authentication, growth curve analysis, and contamination testing (multiplex PCR, bacterial/fungal cultures, Hoechst staining) were performed.
• Population doubling time (PDT) was calculated for the established cell lines.

Main Results:

• Successfully established 51 viable Caspian horse fibroblast cell lines.
• Determined a population doubling time (PDT) of 23 ± 0.5 hours for all cell lines.
• Confirmed the absence of interspecies contamination and microbial contamination through rigorous testing.

Conclusions:

• The established Caspian horse cell lines represent a valuable resource for genomic, post-genomic, and somatic cloning research.
• These cell lines are essential for preserving the genetic diversity of the critically endangered Caspian horse.
• The creation of an accessible cell line database will support future research and conservation initiatives.