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eSymposia | Tissue Plasticity: Preservation and Alteration of Cellular Identity


Invitro spinal disc culture


Oct 5, 2020 12:00am ‐ Oct 5, 2020 12:00am

Description

Invitro spinal disc culture Authors: Nagpal P.S*1 (PhD); Chhabra H.S (MS ortho)2 ; Singh A3. ; Aggarwal, N1. Panda A.(PhD)3; ; Nanda A. 2 ; Mahopatra B. 2 ; Gururaj M. 2 ; Das K. 2; Bansal M. 2; Tandon V 2 ; Address: 1 Dept of Basic Research, ISIC , Vasnt Kunj, New Delhi, 2 Spine Unit, ISIC, Vasnt Kunj, New Delhi 3 PDC-II, National Institute of immunology , Aruna asif ali marg, New Delhi * Presenting Author STUDY DESIGN: Ex vivo in vitro study evaluation of intervertebral disc culture system. OBJECTIVES: To evaluate the feasibility of developing three dimensional intervertebral disc culture model to study disc degeneration in vitro. Key Words: IVD, 3D culture, oestrogenic differentiation Introduction: With current in vivo models for the study of disc and endplate degeneration, it remains difficult to investigate the complex disc metabolism and signalling cascades. Therefore, there is ever growing demand to create three dimensional in vitro model to evaluate the pathologies in a structurally intact organ. METHODS: Cells isolated from 8 human intervertebral disc units were successfully cultured in multi-well plates. Their cell viability, metabolic activity, cellular morphology was observed microscopically. Cells grown in 2 dimensional setup was transferred into 3D scaffolds and their morphology was evaluated by Scanning electron microscopy. RESULTS: Viable cells were successfully isolated from human disc and grown successfully till 8 passages and maintained as active culture for more than 2 months. Actively growing culture at 4th passage was transferred successfully into three dimensional PDLLA-PLGA and chitosan scaffolds. Scanning electron microscopy images confirms their viability and growth in scaffolds. Isolated cells enriched in 2 dimensional culture has the ability to differentiate to osteocytes when activated using oestrogenic media. CONCLUSIONS: Intervertebral cells isolated from human degenerative disc can be enriched invitro into 2 dimensional and three dimensional culture to prepare in vitro disc model. (IEC Ref no: ISIC/RP/2017/71)

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