Identification of Rabbit Annulus Fibrosus-Derived Stem Cells

Abstract

Annulus fibrosus (AF) injuries can lead to substantial deterioration of intervertebral disc (IVD) which characterizes degenerative disc disease (DDD). However, treatments for AF repair/regeneration remain challenging due to the intrinsic heterogeneity of AF tissue at cellular, biochemical, and biomechanical levels. In this study, we isolated and characterized a sub-population of cells from rabbit AF tissue which formed colonies in vitro and could self-renew. These cells showed gene expression of typical surface antigen molecules characterizing mesenchymal stem cells (MSCs), including CD29, CD44, and CD166. Meanwhile, they did not express negative markers of MSCs such as CD4, CD8, and CD14. They also expressed Oct-4, nucleostemin, and SSEA-4 proteins. Upon induced differentiation they showed typical osteogenesis, chondrogenesis, and adipogenesis potential. Together, these AF-derived colony-forming cells possessed clonogenicity, self-renewal, and multi-potential differentiation capability, the three criteria characterizing MSCs. Such AF-derived stem cells may potentially be an ideal candidate for DDD treatments using cell therapies or tissue engineering approaches.

The presence of stem cells in potential stem cell niches of the intervertebral disc region: an in vitro study on rats

“…This result is in accordance with previous in vivo studies of labeling slow cycling cells in the ISN region [21,23,24], further demonstrating the existence of stem cells in the potential ISN. Cell proliferation, as determined by the MTT assay, presented an S-shaped growth curve of both ISN-SCs and BMSCs with a logarithmic growth phase, similar to that of stem cells derived from other tissues as described in previous studies [20,29], suggesting a strong proliferation ability of ISN-SCs, though a better results was observed in BMSCs. NANOG, SOX-2, and OCT-4 are transcription factors that cooperatively maintain the regulatory system of cell self-renewal and pluripotency [42].…”

“…Furthermore, quantitative analysis for comparing the extent of staining between induced and control groups demonstrated strong multilineage differentiation efficiencies of ISN-SCs and BMSCs. These were further confirmed by qPCR analysis, which revealed significantly increased expression of multipotential differentiation-related genes in the induced groups, including osteogenesis (Runx2, OPN, and OCN)-, chondrogenesis (SOX-9, COL2a1 and ACAN)-and adipogenesis (PPARc and C/EBPa)-related genes that were universally used in previous studies on the multilineage differentiation of stem cells [18,20]. Particularly, both of quantitative analysis for comparing the extent of staining and qPCR analysis demonstrated more powerful osteogenic and chondrogenic abilities of ISN-SCs than those of BMSCs.…”

“…This similar promoting migration to repair (PMR) effect has also been observed in many other tissues [13][14][15][16]. Until now, stem cells derived from various parts of the IVD such as the nucleus pulposus (NP), annulus fibrosus (AF), and cartilage endplate (CEP) have been isolated and characterized [17][18][19][20], whereas little is known about endogenous stem cell migration with respect to difficulties in labeling target stem cells in vivo.…”

The Differential Effects of Leukocyte-Containing and Pure Platelet-Rich Plasma on Nucleus Pulposus-Derived Mesenchymal Stem Cells: Implications for the Clinical Treatment of Intervertebral Disc Degeneration

“…Spleen cells were isolated according to the method described previously [32]. Briefly, the harvested spleen tissue from the rabbits above was minced and filtrated with 70  μ m cell strainer (Corning, catalog no.…”

State of the art and future of stem cells in spine

“…A few other studies also identified the stem cells from NP tissue and cartilage endplate, and these stem cells are comparable to bone marrow MSCs in terms of cell surface marker, differentiation capabilities. [31][32][33][34][35][36] Sakai et al 37 recently identified progenitor cells from mice and human NP cells that are Tie2 and disialoganglioside 2 (GD2) positive. The proportion of Tie2 positive cells decreased with aging and disc degeneration.…”