Culture, characterization and differentiation of cells from buffalo (Bubalus bubalis) amnion

Abstract

Stem cells present an important tool in livestock assisted reproduction and veterinary therapeutic field such as tissue engineering. We report for the first time isolation of pluripotent stem cell-like cells expressing pluripotency markers (alkaline phospahatase, OCT-4, NANOG and SOX-2) from the amnion of water buffalo (Bubalus bubalis). The cells showed no apparent abnormalities in their chromosomal profiles before and after cryopreservation. The cytochemical staining revealed that pluripotent cells were capable of undergoing directed differentiation in vitro into osteocytes. It could be inferred that amnionderived pluripotent stem cell-like cells can be isolated, cultured for many passages and differentiated into mesoderm lineage, and may be an alternative source to mesenchymal stem cells. These cells can have applications in assisted reproduction, developmental biological and regenerative medicine.

Buffalo (Bubalus bubalis) term amniotic-membrane-derived cells exhibited mesenchymal stem cells characteristics in vitro

“…Chromosomal spreads at passage 5 and passage 10 revealed that these cells had normal number of chromosomes (2n=50) indicating no apparent changes in ploidy during culture. Previous studies also reported a stable karyotype in cells derived from early buffalo gestation placental tissues (Yadav et al 2011;Dev et al 2012;Mann et al 2013;Singh et al 2013) and term amnion in equine (Seo et al 2013).…”

“…In buffalo, Mann et al (2013) reported that cells cultured from AM of gravid uteri of first trimester collected from slaughter house have positive expression of pluripotency markers (alkaline phosphatase, OCT-4, NANOG and SOX-2) and have an osteogenic differentiation potential. An advantage of the present work over Mann et al (2013) is the isolation of somatic cells from term placentae of known pedigree buffalo and gestational stage of the studied amniotic membranes.…”

“…An advantage of the present work over Mann et al (2013) is the isolation of somatic cells from term placentae of known pedigree buffalo and gestational stage of the studied amniotic membranes. Considering this in mind, the main objective of this study was the isolation, culture and characterization of mesenchymal stem cells of amniotic membrane of term placentae.…”

Membrana amniótica: fonte alternativa de células-tronco mesenquimais em diversas espécies animais

“…Em búfalos, estudos demonstraram que MSCs derivadas do âmnio podem ser isoladas e propagadas em cultivo com várias passagens, podendo ter aplicações em diversas áreas como reprodução assistida, desenvolvimento bioló-gico e medicina regenerativa (Yadav et al 2011, Mann et al 2013. Além disso, foi relatado que células derivadas da membrana amniótica expressam pluripotência e marcadores de células-tronco mesenquimais, e são propensos a se diferenciarem em células de linhagem mesenquimal com diferenciação in vitro (Ghosh et al 2015).…”

Stem Cells from Foetal Adnexa and Fluid in Domestic Animals: An Update on Their Features and Clinical Application

“…,b,c; Mann et al. ), AMSCs expressed NANOG at RT‐PCR, and canine AMSCs were negative (Filioli Uranio et al. , ).…”