Derivation and characterization of sheep bone marrow-derived mesenchymal stem cells induced with telomerase reverse transcriptase

Abstract

Bone marrow mesenchymal stem cells (BMSCs) are a type of adult stem cells with a wide range of potential applications. However, BMSCs have a limited life cycle under normal culturing conditions, which has hindered further study and application. Many studies have confirmed that cells modified by telomerase reverse transcriptase (TERT) can maintain the ability to proliferate over a long period of time. In this study, we constructed a gene expression vector to transfer TERT into sheep BMSCs, and evaluated whether the TERT cell strain was successfully transferred. The abilities of cell proliferation and differentiation were evaluated using the methods including growth curve determination, inheritance stability analysis, multi-directional induction and so on, and the results showed that the cell strain can be cultured to 40 generations, with a normal karyotype rate maintained at 88.24%, and that the cell strain can be transferred and differentiated into neurocytes and lipocytes, proving that it retains the multi-directional transdifferentiation ability.

Smad7-overexpressing rat BMSCs inhibit the fibrosis of hepatic stellate cells by regulating the TGF-β1/Smad signaling pathway

“…Bone marrow cells were harvested by carefully washing the marrow cavity with PBS. The cells were isolated using density gradient centrifugation (×400 g for 10 min at 37°C) and an adherence screening approach ( 10 ). Subsequently, cells were cultured in Dulbecco's modified Eagle's medium (DMEM)/F12 medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Hyclone; GE Healthcare Life Sciences, Logan, UT, USA) at a density of 1×10 6 cells/ml.…”

Mesenchymal Stem Cells for Spinal Cord Injury: Current Options, Limitations, and Future of Cell Therapy

“…These cells are found within the adult bone marrow, where they contribute to hematopoiesis and bone regeneration. BM-MSCs can not only be obtained from humans, rodents, or primates, but also from several animal species such as sheep, dogs, cats, and bovines (Figure 3) [88,92,93,94,95,96,97,98].…”

Small Ruminants and Its Use in Regenerative Medicine: Recent Works and Future Perspectives

“…A caprine model of acute central cervical spinal cord injury syndrome combined with chronic injury was created [ 70 ], and goats were also used to study the effects of different types of laminectomies on spinal cord injury subsequent to acute spinal shorting [ 71 ] and the effects of adipose stem cells seeded on a radiolucent cage filler in spinal fusion [ 72 ]. Particularly in intervertebral disc disease, the application of a gelatin sponge rich in goat bone marrow MSCs (gBM-MSCs) and platelet-rich plasma promoted healing, with histological evidence between 3 to 12 weeks [ 73 ], and other studies on the use of MSCs and basement membrane molecules appear to have potential for cartilage regeneration and chondrogenesis of the nucleus pulposus [ 74 ].…”

“…Karyotypes remain stable up to 20 passages [ 168 ] and cells can have steady heredity and viability up to 48 passages, with less stability recorded in cells harvested from fetuses [ 169 ]. Genetic instability begins to be seen after extensive culturing, probably related to a shortening of telomeres and aging [ 73 ]. The tridifferentiation achieved from different types of chemical compounds allows osteodifferentiation, chondrodifferentiation and adipodifferentiation [ 170 ].…”