title: A Novel Organ Culture Model of Mouse Intervertebral Disc Tissues.
abstract: The intervertebral disc (IVD) is a fibrocartilaginous joint between two vertebral bodies. An IVD unit consists of a gelatinous central nucleus pulposus, encased by the annulus fibrosus, which is sandwiched between cartilaginous endplates (EPs). The IVD homeostasis can be disrupted by injuries, ageing and/or genetic predispositions, leading to degenerative disc disorders and subsequent lower back pain. The complex structure and distinct characteristics of IVDs warrant the establishment of robust in vitro IVD organ culture for studying the etiology and treatment of disc degeneration. Here, we isolate mouse lumbar IVDs and culture the minimal IVD units in submersion or suspension medium supplemented with 2% bovine serum or 10% fetal bovine serum (FBS). We find the minimal IVD units remain healthy for up to 14 days when cultured in submersion culture supplemented with 10% FBS. New bone formation in the EPs of the cultured IVDs can be assessed with calcein labeling. Furthermore, the cultured IVDs can be effectively transduced by recombinant adenovirus, and transgene expression lasts for 2 weeks. Thus, our findings demonstrate that the optimized IVD organ culture system can be used to study IVD biology and screen for biological factors that may prevent, alleviate and/or treat disc degeneration.
authors.forename: Zhengjian
authors.initials: Z
authors.lastname: Yan
authors.forename: Liangjun
authors.initials: L
authors.lastname: Yin
authors.forename: Zhongliang
authors.initials: Z
authors.lastname: Wang
authors.forename: Jixing
authors.initials: J
authors.lastname: Ye
authors.forename: Zhonglin
authors.initials: Z
authors.lastname: Zhang
authors.forename: Ruifang
authors.initials: R
authors.lastname: Li
authors.forename: Sahitya K
authors.initials: SK
authors.lastname: Denduluri
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Qiang
authors.initials: Q
authors.lastname: Wei
authors.forename: Lianggong
authors.initials: L
authors.lastname: Zhao
authors.forename: Shun
authors.initials: S
authors.lastname: Lu
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Shengli
authors.initials: S
authors.lastname: Tang
authors.forename: Lewis L
authors.initials: LL
authors.lastname: Shi
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Lee
authors.forename: Tong-Chuan
authors.initials: TC
authors.lastname: He
authors.forename: Zhong-Liang
authors.initials: ZL
authors.lastname: Deng
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D057809:HEK293 Cells; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology*; D055959:Intervertebral Disc Degeneration / Q000628:therapy; D008161:Lumbosacral Region / Q000502:physiology; D008297:Male; D051379:Mice; D009924:Organ Culture Techniques / Q000379:methods*; D018809:Proliferating Cell Nuclear Antigen / Q000096:biosynthesis; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D018809:Proliferating Cell Nuclear Antigen