title: A Substance Exchanger-Based Bioreactor Culture of Pig Discs for Studying the Immature Nucleus Pulposus.
abstract: Various research models have been developed to study the biology of disc cells. Recently, the adult disc nucleus pulposus (NP) has been well studied. However, the immature NP is underinvestigated due to a lack of a suitable model. This study aimed to establish an organ culture of immature porcine disc by optimizing culture conditions and using a self-developed substance exchanger-based bioreactor. Immature porcine discs were first cultured in the bioreactor for 7 days at various levels of glucose (low, medium, high), osmolarity (hypo-, iso-, hyper-) and serum (5, 10, 20%) to determine the respective optimal level. The porcine discs were then cultured under the optimized conditions in the novel bioreactor, and were compared with fresh discs at day 14. For high-glucose, iso-osmolarity, or 10% serum, cell viability, the gene expression profile (for anabolic genes and catabolic genes), and glycosaminoglycan (GAG) and hydroxyproline (HYP) contents were more favorable than for other levels of glucose, osmolarity, and serum. When the immature discs were cultured under the optimized conditions using the novel bioreactor for 14 days, the viability of the immature NP was maintained based on histology, cell viability, GAG and HYP contents, and matrix molecule expression. In conclusion, the viability of the immature NP in organ culture could be maintained under the optimized culture conditions (high-glucose, iso-osmolarity, and 10% serum) in the substance exchanger-based bioreactor.
authors.affiliation: Department of Orthopedic Surgery, Southwest Hospital, Third Military Medical University.
authors.forename: Pei
authors.initials: P
authors.lastname: Li
authors.affiliation: Department of Orthopedic Surgery, Southwest Hospital, Third Military Medical University.
authors.forename: Yibo
authors.initials: Y
authors.lastname: Gan
authors.affiliation: Department of Orthopedic Surgery, Chongqing Three Gorges Central Hospital.
authors.forename: Haoming
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Orthopedic Surgery, Xinqiao Hospital, Third Military Medical University, Chongqing, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Orthopedic Surgery, Southwest Hospital, Third Military Medical University.
authors.forename: Lei
authors.initials: L
authors.lastname: Song
authors.affiliation: Department of Orthopedic Surgery, Southwest Hospital, Third Military Medical University.
authors.forename: Liyuan
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Orthopedic Surgery, Southwest Hospital, Third Military Medical University.
authors.forename: Bin
authors.initials: B
authors.lastname: Ouyang
authors.affiliation: Department of Orthopedic Surgery, Southwest Hospital, Third Military Medical University.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D000818:Animals; D019149:Bioreactors; D002470:Cell Survival; D003470:Culture Media / Q000378:metabolism; D004867:Equipment Design; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D005947:Glucose / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism; D006909:Hydroxyproline / Q000378:metabolism; D008297:Male; D045726:Metalloproteases / Q000235:genetics / Q000378:metabolism; D000070614:Nucleus Pulposus / Q000166:cytology / Q000502:physiology*; D009924:Organ Culture Techniques / Q000295:instrumentation* / Q000379:methods; D009994:Osmolar Concentration; D044968:Regenerative Medicine / Q000295:instrumentation* / Q000379:methods; D034421:Sus scrofa; D013997:Time Factors; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods
chemical_list: D003470:Culture Media; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D045726:Metalloproteases; D005947:Glucose; D006909:Hydroxyproline
keywords: -Bioreactor; -Degeneration; -Immature; -Nucleus pulposus; -Organ culture; Intervertebral disc