Abstract
Lower back pain is a leading cause of disability worldwide. The recovery of nucleus pulposus (NP) progenitor cells (NPPCs) from the intervertebral disc (IVD) holds high promise for future cell therapy. NPPCs are positive for the angiopoietin-1 receptor (Tie2) and possess stemness capacity. However, the limited Tie2+ NPC yield has been a challenge for their use in cell-based therapy for regenerative medicine. In this study, we attempted to expand NPPCs from the whole NP cell population by spheroid-formation assay. Flow cytometry was used to quantify the percentage of NPPCs with Tie2-antibody in human primary NP cells (NPCs). Cell proliferation was assessed using the population doublings level (PDL) measurement. Synthesis and presence of extracellular matrix (ECM) from NPC spheroids were confirmed by quantitative Polymerase Chain Reaction (qPCR), immunostaining, and microscopy. Compared with monolayer, the spheroid-formation assay enriched the percentage of Tie2+ in NPCs’ population from ~10% to ~36%. Moreover, the spheroid-formation assay also inhibited the proliferation of the Tie2- NPCs with nearly no PDL. After one additional passage (P) using the spheroid-formation assay, NPC spheroids presented a Tie2+ percentage even further by ~10% in the NPC population. Our study concludes that the use of a spheroid culture system could be successfully applied to the culture and expansion of tissue-specific progenitors.
Highlights
The disability induced by lower back pain (LBP) is a global issue with a substantial social burden [1]
Results the implication of NP cells (NPCs) spheroids formed on an ultra-low-attachment surface
Fceulrltsh[e2r6m].ore, spheroid-formation culture has becTohmee tihmepmliaciantisotrnesamofwasyphtoercouildtu-froermneautrioalnsteamssacyellsa[r2e6]i.nvolved in many different types of stemTheciemllsplifcraotmionsotohfesrphteisrsouide-sf.orTmharetieonimaspsaoyrtaarnetinavsoplevcetds inarme akneyydiifnfersetnetmtypceesllofrsetseemarcchel:ls isofrloamtioont,hperrotliisfseuraetsi.oTnh, raenedimmpaionrtteannatnacsepeocfttshaerier ksteeyminnesstse.mTcheellsrpehseeraoricdh-:foisromlaattiioonn, pasrsoaliyfeirsaatilorena,daynd wmidaeilnytuenseadncienocfatnhceeirr sstteemmnceesllss. iTshoelastpiohnerooriden-froircmhmateionnt [a2s5s,a3y7]i,sleatlhreaaldsycrweeindesluybutysepde i[n37c]a, nacnedr tshteem drcuegll-sreissoislatatinotnsuorbteynpreic[h3m8]e.nTtw[2o5,r3e7c]e, nlettshtauldsicerseeshnoswubedtypthea[t3s7p],haenrodidthfeordmruagti-orensicsotaunldt spurbottyepctet[h3e8]. mTuwltioporteecnecnyt osftuaddiipesosseh-doewreivdedthsatet mspcheellrso[i2d4,f3o9r]m
Summary
The disability induced by lower back pain (LBP) is a global issue with a substantial social burden [1]. The main reason for LBP, apart from trauma to the intervertebral disc (IVD), is the degeneration of the intervertebral disc (IVDD) caused by aging or genetic predisposition [2]. Disability induced by IVDD means a reduction in the labor force participation rate and a tremendous decrease in life quality for patients. The IVD consists of three central tissues: (i) the nucleus pulposus (NP), (ii) the annulus fibrosus (AF), and (iii) the cartilaginous endplate (CEP) [4]. The health of the IVD depends on the well-being of all three tissue components [5]. If only one of the three tissues partly degenerates, it will affect the health status of the entire IVD
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
