title: Potential of co-culture of nucleus pulposus mesenchymal stem cells and nucleus pulposus cells in hyperosmotic microenvironment for intervertebral disc regeneration.
abstract: Nucleus pulposus mesenchymal stem cells (NPMSCs) are a potential cell source for intervertebral disc (IVD) regeneration, but little is known about their response to IVD-like high osmolarity (400 mOsm). This study was to investigate the viability, proliferation and protein biosynthesis of nucleus pulposus cells (NPCs), NPMSCs and co-cultured NPMSCs-NPCs under IVD-like high osmolarity conditions. NPCs and NPMSCs were isolated and cultured under standard and IVD-like high osmolarity conditions for 1 or 2 weeks. Cell viability was measured by annexin V-FITC and PI staining, and cell proliferation measured by MTT assay. The expression of SOX-9, aggrecan and collagen-II was measured by RT-PCR and Western blot analyses. IVD-like high osmolarity condition slightly inhibited cell viability and decreased the expression of SOX-9, aggrecan and collagen-II at the mRNA and protein levels in all groups compared with standard condition. NPMSCs could tolerate IVD-like high osmolarity, and NPCs-NPMSCs co-culture increased cell proliferation and the expression of SOX-9, aggrecan and collagen-II under both culture conditions, suggesting that co-culture of NPMSCs-NPCs has potential application for IVD regeneration.
authors.affiliation: Department of Orthopedics, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, No. 88 Jie Fang Road, Hangzhou, 310009, China.
authors.forename: Yi-Qing
authors.initials: YQ
authors.lastname: Tao
authors.forename: Cheng-Zhen
authors.initials: CZ
authors.lastname: Liang
authors.forename: Hao
authors.initials: H
authors.lastname: Li
authors.forename: Yu-Jie
authors.initials: YJ
authors.lastname: Zhang
authors.forename: Fang-Cai
authors.initials: FC
authors.lastname: Li
authors.forename: Gang
authors.initials: G
authors.lastname: Chen
authors.forename: Qi-Xin
authors.initials: QX
authors.lastname: Chen
mesh_terms: D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D007403:Intervertebral Disc / Q000473:pathology / Q000502:physiology*; D055959:Intervertebral Disc Degeneration / Q000473:pathology / Q000628:therapy; D008297:Male; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D009994:Osmolar Concentration; D014176:Protein Biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012038:Regeneration; D044968:Regenerative Medicine; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism
chemical_list: D053674:Aggrecans; D024043:Collagen Type II; D055758:SOX9 Transcription Factor
keywords: co-culture; intervertebral disc; mesenchymal stem cell; nucleus pulposus; osmolarity