Successful fishing for nucleus pulposus progenitor cells of the intervertebral disc across species

• DOI: 10.1002/jsp2.1018
• Publication Date: 2018-06-11T09:50:36Z
• Authors: Daisuke Sakai; Jordy Schol; Frances C. Bach; Adel Tekari; Nobuho Sagawa; Yoshihiko Nakamura; Samantha C.W. Chan; Tomoko Nakai; Laura B. Creemers; Daniela A. Frauch...; Rahel D. May; Sibylle Grad; Masahiko Watanabe; Marianna A. Tryfo...; Benjamin Gantenbein
• Keywords: Cell Sorting; Angiopoietin receptor

Background Recently, Tie2/TEK receptor tyrosine kinase (Tie2 or syn. angiopoietin‐1 receptor) positive nucleus pulposus progenitor cells were detected in human, cattle, and mouse. These show remarkable multilineage differentiation capacity direct correlation with intervertebral disc (IVD) degeneration are therefore an interesting target for regenerative strategies. Nevertheless, there remains controversy over the presence function of these Tie2 + (NPCs), part due to difficulty identification isolation. Purpose Here, we present a comprehensive protocol sorting NPCs from canine, bovine, murine IVD tissue. We describe enhanced conditions expansion optimized fluorescence‐activated cell sorting‐based methodology sort analyze NPCs. Methods flow cytometry protocols isolate population aforementioned species. Moreover, crucial pitfalls prevent loss during isolation process. A cross‐species phylogenetic analysis across species is presented. Results Our efficient towards labeling The total procedure requires approximately 9 hours, 4 16 can take up multiple weeks, dependent on application, age, disease state, Phylogenetic TEK gene revealed strong homology among Conclusions Current could be confirmed mouse, human specimens. presented successfully multipotent cells. biological isolated based expression needs by functional assays such as vitro differentiation. culture maintain their possible proliferation fraction subject future research.