Animal Reproduction (AR)
https://www.animal-reproduction.org/article/5b5a6069f7783717068b474e
Animal Reproduction (AR)
Original Article

Isolation and characterization of mesenchymal cells isolated from caprine umbilical cord matrix

M. Moshrefi, H. Babaei, S.N. Nematollahi-Mahani

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Abstract

To date, no investigations have been found about the isolation and in vitro cell culture characterization of Wharton's jelly mesenchymal cells (WJMC) from caprine umbilical cord (CUC). Therefore, in the present experiment the isolation protocol and growth kinetic of CUC matrix cells were studied. CUC were collected from an abattoir and pregnant uteri (lategestation) and their Wharton's jellies (WJ) were cut into 2 × 2 mm2 fragments for explanting. Explants (n = 8-10) were transferred to each 35 mm culture dish. WJ explants were removed 5 days after plating and the remaining adherent cells were cultured for another 5 days. CUC isolated cells were immunostained for Actin protein and histochemically were assayed for the presence of alkaline phosphatase (AP) activity. Besides, in this study growth kinetic and clonogenicity were evaluated for the isolated cells. CUC isolated cells displayed spindle-form and small round-shape with large nucleus. Confluent cells formed colonies that presented AP activity. Immunocytochemical analysis revealed expression of Actin protein. Initial seeding concentration of 2 × 104 cells resulted in smaller time for doubling the population compared to fetal fibroblasts (46.6 vs. 54.3 h, respectively). In conclusion, WJ of CUC contains an easily isolated and rich source of myofibroblast-like cells which exhibit some stem cell-like properties.

Keywords

actin, alkaline phosphatase, caprine, mesenchymal cells, umbilical cord
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