Optimization of Spheroid Colony Culture and Cryopreservation of Nucleus Pulposus Cells for the Development of Intervertebral Disc Regenerative Therapeutics

After the discovery of functionally superior Tie2-positive nucleus pulposus (NP) progenitor cells, new methods were needed to enable mass culture and cryopreservation maintain these cells in an undifferentiated state with high cell yield. We used six types EZSPHERE® dishes, which support spheroid-forming colony culture, examined NP spheroid-formation ability, number, proliferation, mRNA expression ACAN, COL1A2, COL2A1, ANGPT1. Six different solutions for potential use clinical by comparing effects exposure time during on viability, Tie2-positivity, proliferation rates. The spheroid formation rate was 45.1% 7.75 times using dishes. levels COL2A1 ANGPT1 also high. In cryopreservation, CryoStor10 (CS10) produced ?90% viability a after thawing. CS10 had 12.6% culturing 5 days These results suggest that EZSPHERE enabled without hydrogel products is best medium retaining phenotype viability. Together, data provide useful information cell-based therapeutics clinic.