Buffalo (Bubalus bubalis) ES Cell–Like Cells are Capable of In Vitro Skeletal Myogenic Differentiation. Issue 2 (12th July 2012)

• Record Type: Journal Article
• Authors: Singh, MK; Singh, KP; Kumar, D; Shah, RA; Anand, T; Chauhan, MS; Manik, RS; Singla, SK; Palta, P
• Is Part Of: Reproduction in domestic animals. Volume 48:Issue 2(2013)
• Page Start: 284
• Page End: 291
• Publication Date: 2012-07-12
• Subjects: Animal breeding -- Periodicals; Veterinary pathology -- Periodicals; Livestock -- Breeding -- Periodicals; 636.082
• DOI: 10.1111/j.1439-0531.2012.02146.x
• Languages: English
• ISSNs: 0936-6768

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Contents</title> <p>When buffalo embryonic stem (ES) cell–like cells that expressed surface markers SSEA‐4, TRA‐1‐60, TRA‐1‐81, CD9 and CD90 and intracellular markers OCT4, SOX2 and FOXD3, as shown by immunofluorescence, and that expressed <italic>REX‐1</italic> and <italic>NUCLEOSTEMIN</italic> as confirmed by RT‐PCR, were subjected to suspension culture in hanging drops in absence of LIF and buffalo foetal fibroblast feeder layer support, they differentiated to form three‐dimensional embryoid bodies (EBs). Of 231 EBs examined on Day 3 of suspension culture, 141 (61.3 ± 3.09%) were of compact type, whereas 90 (38.4 ± 3.12%) were of cystic type. The cells obtained from EBs were found to express <italic>NF‐68</italic> and <italic>NESTIN</italic> (ectodermal lineage), <italic>BMP‐4</italic> and α‐skeletal actin (mesodermal lineage), and α‐fetoprotein, <italic>GATA‐4</italic> and <italic>HNF‐4</italic> (endodermal lineage). When these EBs were cultured on gelatin‐coated dishes, they spontaneously differentiated to several cell types such as epithelial‐ and neuron‐like cells. When EBs were cultured in the presence of 1 or 2% DMSO or 10<sup>−8 </sup><sc>m</sc> or 10<sup>−7 </sup><sc>m</sc> retinoic acid for 25 days, ES cells could be directed to form muscle cell–like cells, the identity of which was confirmed by expression of α‐actinin by immunofluorescence and of <italic>MYF‐5, MYOD</italic> and <italic>MYOGENIN</italic> genes by RT‐PCR. <italic>MYOD</italic> was first detected on Day 10 in both treatment groups and on Day 15 in controls, whereas <italic>MYOGENIN</italic> was first detected on Day 10, Day 15 and Day 25 in the presence of retinoic acid, in the presence of DMSO and in controls, respectively. The present study demonstrates the ability of buffalo ES cell–like cells to undergo directed differentiation to cells of skeletal myogenic lineage.</p> </abstract>