title: Sox-9 facilitates differentiation of adipose tissue-derived stem cells into a chondrocyte-like phenotype in vitro.
abstract: The purpose of this study is to test whether ectopic expression of Sox-9 can induce adipose tissue-derived stem cells (ASCs) to function as real nucleus pulposus (NP) cells in vitro. Adenoviral vectors expressing Sox-9 were reported to infect the chondroblastic and human disc cells, which resulted in increased Sox-9 and type II collagen production. ASCs were isolated from rat inguinal adipose pad, characterized, and transduced in vitro with a retroviral vector encoding the Sox-9 gene. Sox-9-engineered ASCs (ASCs/Sox-9) were induced for the chondrocyte-like cell differentiation by 3D cultured in alginate beads and TGF-β3 for 2 weeks. Expression of exogenous Sox-9 protein was detected. Type II collagen and Aggrecan gene expressions of induced ASCs/Sox-9 were measured using real-time PCR; proteoglycans expressions were measured by checking the glycosaminoglycan content and type II collagen production by enzyme-linked immunosorbent assay. Isolated ASCs were CD 29(+) /CD44(+) /C-Kit(-) /Lin(-) /CD34(-) /CD45(-) . ASCs/Sox-9 expressed marked increase in exogenous Sox-9 protein. After induction, type II collagen gene expression was sevenfold higher in mRNA levels, with an approximately twofold increase in protein levels of ASCs/Sox-9 compared to ASCs. Type II collagen and proteoglycan productions were significantly increased in the ASCs/Sox-9 compared to the ASCs. In addition, co-culture of induced ASCs/Sox-9 with matured NP cells resulted in enhanced increase in proteoglycan and type II collagen production. Constitutive retroviral expression of Sox-9 could efficiently induce ASCs differentiation into chondrocyte-like cells. This novel approach may provide a practicable system for a simple and rapid differentiation of ASCs into chondrocyte-like cells which may be potentially used as a stem cell-based therapeutic tool for the treatment of degenerative disc diseases.
authors.affiliation: Department of Anesthesiology, University of Miami Miller School of Medicine, Miami, Florida 33136, USA.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Yang
authors.forename: Chun-Yuh Charles
authors.initials: CY
authors.lastname: Huang
authors.forename: Keith A
authors.initials: KA
authors.lastname: Candiotti
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Zeng
authors.forename: Taiyi
authors.initials: T
authors.lastname: Yuan
authors.forename: Jinliang
authors.initials: J
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Yu
authors.forename: Salahadin
authors.initials: S
authors.lastname: Abdi
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D053687:Adult Stem Cells / Q000378:metabolism*; D053674:Aggrecans / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D019902:Chondrocytes / Q000166:cytology*; D018920:Coculture Techniques; D024043:Collagen Type II / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D007403:Intervertebral Disc / Q000166:cytology; D010641:Phenotype; D051381:Rats; D012190:Retroviridae; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D053782:Transforming Growth Factor beta3 / Q000378:metabolism
chemical_list: D053674:Aggrecans; D024043:Collagen Type II; D016326:Extracellular Matrix Proteins; D055758:SOX9 Transcription Factor; D053782:Transforming Growth Factor beta3