Isolation and characterization of buffalo (<i>bubalus bubalis</i>) amniotic mesenchymal stem cells derived from amnion from the first trimester pregnancy

Abstract

Amniotic mesenchymal stem cells (AMSCs) from livestock are valuable resources for animal reproduction and veterinary therapeutic. The purpose of this study is to explore a suitable way to isolate and culture the buffalo AMSCs (bAMSCs), and to identify their biological characteristics. Digestion with a combination of trypsin-EDTA and collagenase type I could obtain pure bAMSCs more effectively than trypsin-EDTA or collagenase type I alone. bAMSCs could proliferate steadily in vitro culture and exhibited fibroblastic-like morphology in vortex-shaped colony. bAMSCs were positive for MSC-specific markers CD44, CD90, CD105, CD73, β-integrin (CD29) and CD166, and pluripotent markers OCT4, SOX2, NANOG, REX-1, SSEA-1, SSEA-4 and TRA-1-81, but negative for hematopoietic markers CD34, CD45 and epithelial cells specific marker Cytokeratin 18. In addition, bAMSCs were capable of differentiating into adipogenic, osteogenic, chondrogenic and neural lineages, with expression of FABP4, Ost, ACAN, COL2A1, Nestin and β III-tubulin. Glycogen synthase kinase 3 inhibitor: kenpaullone promoted bAMSCs to differentiate into neural lineage. This study provides an effective protocol to obtain and characterize bAMSCs, which have proven useful as a cell resource for buffalo cell reprogramming studies and transgenic animal production.

Producing Human Amniotic Epithelial Cells-only Membrane for Transplantation

“…1G ). These results were according with the former findings in characteristic of hAECs [9,16,38-40]. Therefore, it showed that the cells on hAECs membrane generated here were very likely to be highly pure hAECs.…”

“…To distinctly identify hAECs and hAMSCs, the most accepted biomarkers to distinguish hAECs and hAMSCs were EpCAM (hAECs-positive), CK-8 and CK-18 (hAECs-positive) and α-SMA (hAECs-negative). Results showed that hAMSCs isolated by former established methods had 2.54% EpCAM + result [4,23,38] ( Fig. 1B ) ( Table 1 ).…”

“…1E ). In IF analysis, the hAMSCs isolated by established procedures showed EpCAM − [4,23,38] ( Fig. 1D ).…”

Mesenchymal stem cell: Basic research and potential applications in cattle and buffalo

“…Currently, FBS is used at a variable concentration ranging from 5% to 25%. bufBM‐MSCs grow better at 15% (Gade et al, ) and for cattle 10% concentration appears sufficient (Lu et al, 2014) though 20% has also been used (de Moraes et al, 2014; Deng et al, ). In addition to basic ingredients, cultured MSCs proliferation may be improved with supplementation of basic fibroblast growth factor (bFGF).…”

“…The fibroblastic cells also exhibit better migration abilities and/ homing property (depicted by higher expression of CD44) (Rossi et al, 2014). 3, 6, 9, and 20 passages remain 43.9 ± 2, 45.9 ± 3, 46.7 ± 3, and 60 ± 5 hr, respectively (Deng et al, 2018).…”