Assess The Pluripotency of Caprine Umbilical Cord Wharton’s Jelly Mesenchymal Cells By RT-PCR Analysis of Early Transcription Factor Nanog

Abstract

Objective- In the present study, we investigated the isolation protocol, population doubling time (PDT) and the expression of a pluripotential gene by RT-PCR analysis of early transcription factor Nanog in caprine umbilical cord (CUC) Wharton's jelly mesenchymal cells (WJMCs). Design- Experimental in vitro study. Animals- Four mix breed goat. Procedures- CUCs were collected from abattoir pregnant uteri and their Wharton's jellies (WJs) were cut into 2×2 mm2 segments for explanting. 8-10 segments were explanted into each 35 mm culture dish. WJ explants were removed 5 days after plating and the remaining adherent cells were cultured for another 5 days. Isolated cells were histochemically assayed for the presence of alkaline phosphatase (AP) activity. RT-PCR was used to assess the presence of Nanog mRNA. Besides, in this study the growth kinetic was evaluated for the isolated cells. Results- CUC isolated cells displayed spindle-form and small round-shape with high nucleus. Confluent cells formed colonies that indicated AP activity. Initial seeding concentration of 2 × 104 CUC-WJMCs resulted in shorter PDT compared to fetal fibroblasts (46.57 vs. 54.29 hours, respectively). Expression of Nanog was undetectable in 9th passage of CUC-WJMCs. Conclusion and Clinical Relevance- CUC contains an easily obtainable source of mesenchymal cells which exhibit stem cells properties but probably these cells are not pluripotent.