Li Pei, Gan Yibo, Wang Haoming, Xu Yuan, Song Lei, Wang Liyuan, Ouyang Bin, Zhou Qiang
Department of Orthopedic Surgery, Southwest Hospital, Third Military Medical University.
Department of Orthopedic Surgery, Chongqing Three Gorges Central Hospital.
Artif Organs. 2017 Nov;41(11):E308-E319. doi: 10.1111/aor.12834. Epub 2017 Feb 11.
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.
已经开发了各种研究模型来研究椎间盘细胞的生物学特性。最近，成人椎间盘髓核（NP）已得到充分研究。然而，由于缺乏合适的模型，未成熟的NP研究不足。本研究旨在通过优化培养条件并使用自行开发的基于物质交换器的生物反应器建立未成熟猪椎间盘的器官培养。首先将未成熟猪椎间盘在生物反应器中于不同水平的葡萄糖（低、中、高）、渗透压（低渗、等渗、高渗）和血清（5%、10%、20%）下培养7天，以确定各自的最佳水平。然后将猪椎间盘在新型生物反应器中于优化条件下培养，并在第14天与新鲜椎间盘进行比较。对于高糖、等渗或10%血清，细胞活力、基因表达谱（合成代谢基因和分解代谢基因）以及糖胺聚糖（GAG）和羟脯氨酸（HYP）含量比其他葡萄糖、渗透压和血清水平更有利。当使用新型生物反应器在优化条件下将未成熟椎间盘培养14天时，基于组织学、细胞活力、GAG和HYP含量以及基质分子表达，未成熟NP的活力得以维持。总之，在基于物质交换器的生物反应器中，在优化培养条件（高糖、等渗和10%血清）下，未成熟NP在器官培养中的活力可以得到维持。
