Rat spinal motion segment in organ culture : A cell viability study

• Published in: Spine (Philadelphia, Pa. 1976) Pa. 1976), 2006-05, Vol.31 (12), p.1291-1298
• Main Authors: LIM, Tae-Hong, RAMAKRISHNAN, Prem S, KURRIGER, Gail L, MARTIN, James A, STEVENS, Jeff W, KIM, Jaehyun, MENDOZA, Sergio A, YOON, S. Tim
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Cell Survival; Cerebrospinal fluid. Meninges. Spinal cord; Development. Senescence. Regeneration. Transplantation; Feasibility Studies; Fundamental and applied biological sciences. Psychology; Injuries of the nervous system and the skull. Diseases due to physical agents; Intervertebral Disc - cytology; Intervertebral Disc - physiology; Lumbar Vertebrae; Medical sciences; Nervous system (semeiology, syndromes); Neurology; Organ Culture Techniques; Rats; Rats, Sprague-Dawley; Time Factors; Tissue Survival; Traumas. Diseases due to physical agents; Vertebrates: nervous system and sense organs
• ISSN: 0362-2436; 1528-1159
• DOI: 10.1097/01.brs.0000218455.28463.f0

This study investigated tissue integrity and viability of cells in an organ culture system of intervertebral disc (IVD) with adjoining vertebral bodies. The goal of this study was to design a methodology to maintain an IVD motion segment in organ culture, thereby preserving viability and tissue architecture. Study of IVD mechanobiology in vitro necessitates availability of vertebral bodies for controlled application of complex loads. IVD motion segments were dissected from rat lumbar segments and maintained in organ culture and cell viability was evaluated histochemically using NitroBlue Tetrazolium. Tissue integrity and morphology were evaluated using conventional histologic techniques. The in vitro organ culture of motion segments maintained the viability and tissue integrity for 14 days. More than 95% viability in all three regions of interest (anulus fibrosus, nucleus pulposus, end plates) was maintained for 14 days in culture. Our initial results suggest that long-term motion segment culture is practical, and the inclusion of vertebral bodies will facilitate anchoring during biomechanical stimulation. Thus, we expect the culture system to provide us with an excellent model for studying the pathomechanics of IVD degeneration and the effects of mechanical stimulation on the biology of IVD cells.