Chapter 4 Methods and Protocols in Peripheral Nerve Regeneration Experimental Research: Part I—Experimental Models

Tos, Pierluigi; Ronchi, Giulia; Papalia, I.; Sallen, Vera; Legagneux, Josette; Geuna, Stefano; Giacobini-Robecchi, Maria G.

doi:10.1016/s0074-7742(09)87004-9

Cited by 80 publications

(91 citation statements)

References 113 publications

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“…To study novel approaches in peripheral nerve reconstruction and nerve graft tissue engineering, the sciatic nerve (SN) is easily accessible and its length provides the opportunity of addressing long gap repair (Tos et al., 2009) with a number of functional tests (Navarro, 2016). The SN model, however, is often inflicted by automutilation behavior and the development of hamstring contractions, eventually reducing the number of animals available for analysis and the significance of the results (Navarro, 2016; Weber, Proctor, Warner, & Verheyden, 1993).…”

Section: Introductionmentioning

confidence: 99%

Reflex‐based grasping, skilled forelimb reaching, and electrodiagnostic evaluation for comprehensive analysis of functional recovery—The 7‐mm rat median nerve gap repair model revisited

2017

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IntroductionThe rat median nerve injury and repair model gets increasingly important for research on novel bioartificial nerve grafts. It allows follow‐up evaluation of the recovery of the forepaw functional ability with several sensitive techniques. The reflex‐based grasping test, the skilled forelimb reaching staircase test, as well as electrodiagnostic recordings have been described useful in this context. Currently, no standard values exist, however, for comparison or comprehensive correlation of results obtained in each of the three methods after nerve gap repair in adult rats.MethodsHere, we bilaterally reconstructed 7‐mm median nerve gaps with autologous nerve grafts (ANG) or autologous muscle‐in‐vein grafts (MVG), respectively. During 8 and 12 weeks of observation, functional recovery of each paw was separately monitored using the grasping test (weekly), the staircase test, and noninvasive electrophysiological recordings from the thenar muscles (both every 4 weeks). Evaluation was completed by histomorphometrical analyses at 8 and 12 weeks postsurgery.ResultsThe comprehensive evaluation detected a significant difference in the recovery of forepaw functional motor ability between the ANG and MVG groups. The correlation between the different functional tests evaluated precisely displayed the recovery of distinct levels of forepaw functional ability over time.ConclusionThus, this multimodal evaluation model represents a valuable preclinical model for peripheral nerve reconstruction approaches.

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Section: Introductionmentioning

confidence: 99%

Reflex‐based grasping, skilled forelimb reaching, and electrodiagnostic evaluation for comprehensive analysis of functional recovery—The 7‐mm rat median nerve gap repair model revisited

2017

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“…Whereas a clear prevalence in the selection of the rat for nerve regeneration studies can still be detected in the literature, recently the availability of a number of genetically modified mouse colonies is causing an increasing use of mice for the experimental investigation of neural repair in the peripheral nervous system . Most mouse studies so far have been based on sciatic nerve repair and regeneration (see Table I in Tos et al, 2009). In a recent paper, we have described the use of the mouse median nerve model for the experimental investigation of end-to-end nerve reconstruction (Tos et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Standardized crush injury of the mouse median nerve

Ronchi

Raimondo

Varejão

et al. 2010

Journal of Neuroscience Methods

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“…Values represent means ± SD calculated from three samples in each group. *P \ 0.05 as compared with SMDF(-) The PC12-IFRS1 coculture model has significant advantages when compared to previous models using primary cultured neurons and/or Schwann cells: (1) it can be prepared at the researchers' convenience without the timeconsuming processes of the primary culture; (2) it consists of pure neuronal and Schwann cell lines and does not include fibroblasts and other cells derived from primary cultures; and (3) it is free of ethical problems that may arise from sacrificing animals, and meets the requirement of the three Rs (Replacement, Reduction, and Refinement) for limiting animal studies (Tos et al 2009). Furthermore, the model can be prepared and maintained by routine culture techniques without genetic manipulation of cells.…”

Section: Discussionmentioning

confidence: 99%

Myelination in coculture of established neuronal and Schwann cell lines

Sango

Kawakami

Yanagisawa

et al. 2012

Histochem Cell Biol

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Establishing stable coculture systems with neuronal and Schwann cell lines has been considered difficult, presumably because of their high proliferative activity and phenotypic differences from primary cultured cells. The present study is aimed at developing methods for myelin formation under coculture of the neural crest-derived pheochromocytoma cell line PC12 and the immortalized adult rat Schwann cell line IFRS1. Prior to coculture, PC12 cells were seeded at low density (3 × 10(2)/cm(2)) and maintained in serum-free medium with N2 supplement, ascorbic acid (50 μg/ml), and nerve growth factor (NGF) (50 ng/ml) for a week. Exposure to such a NGF-rich environment with minimum nutrients accelerated differentiation and neurite extension, but not proliferation, of PC12 cells. When IFRS1 cells were added to NGF-primed PC12 cells, the cell density ratio of PC12 cells to IFRS1 cells was adjusted from 1:50 to 1:100. The cocultured cells were then maintained in serum-free medium with B27 supplement, ascorbic acid (50 μg/ml), NGF (10 ng/ml), and recombinant soluble neuregulin-1 type III (25 ng/ml). Myelin formation was illustrated by light and electron microscopy performed at day 28 of coculture. The stable PC12-IFRS1 coculture system is free of technical and ethical problems arising from the primary culture and can be a valuable tool to study peripheral nerve degeneration and regeneration.

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Chapter 4 Methods and Protocols in Peripheral Nerve Regeneration Experimental Research: Part I—Experimental Models

Cited by 80 publications

References 113 publications

Reflex‐based grasping, skilled forelimb reaching, and electrodiagnostic evaluation for comprehensive analysis of functional recovery—The 7‐mm rat median nerve gap repair model revisited

Reflex‐based grasping, skilled forelimb reaching, and electrodiagnostic evaluation for comprehensive analysis of functional recovery—The 7‐mm rat median nerve gap repair model revisited

Standardized crush injury of the mouse median nerve

Myelination in coculture of established neuronal and Schwann cell lines

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