A systematic evaluation of Schwann cell injection into acellular cold-preserved nerve grafts

Nithya J. Jesuraj, Katherine B. Santosa, Piyaraj Newton, Z. Liu, Daniel A. Hunter, Susan E. Mackinnon, Shelly E. Sakiyama-Elbert, Philip J. Johnson

Research output: Contribution to journalArticle

30 Scopus citations

Abstract

Peripheral nerve regeneration after injury depends on environmental cues and trophic support. Schwann cells (SCs) secrete trophic factors that promote neuronal survival and help guide axons during regeneration. The addition of SCs to acellular nerve grafts is a promising strategy for enhancing peripheral nerve regeneration; however, inconsistencies in seeding parameters have led to varying results. The current work sought to establish a systematic approach to seeding SCs in cold-preserved acellular nerve grafts. Studies were undertaken to (1) determine the needle gauge for optimal cell survival and minimal epineurial disruption during injection, (2) track the seeded SCs using a commercially available dye, and (3) evaluate the seeding efficiency of SCs in nerve grafts. It was determined that seeding with a 27-gauge needle resulted in the highest viability of SCs with the least damage to the epineurium. In addition, Qtracker® dye, a commercially available quantum dot nanocrystal, was used to label SCs prior to transplantation, which allowed visualization of the seeded SCs in nerve grafts. Finally, stereological methods were used to evaluate the seeding efficiency of SCs in nerve grafts immediately after injection and following a 1- or 3-day in vitro incubation in SC growth media. Using a systematic approach, the best needle gauge and a suitable dye for SC visualization in acellular nerve grafts were identified. Seeding efficiency in these grafts was also determined. The findings will lead to improvements ability to assess injection of cells (including SCs) for use with acellular nerve grafts to promote nerve regeneration.

Original languageEnglish
Pages (from-to)209-215
Number of pages7
JournalJournal of Neuroscience Methods
Volume197
Issue number2
DOIs
StatePublished - Apr 30 2011

Keywords

  • Cell transplantation
  • Nerve regeneration
  • Peripheral nerve injury

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    Jesuraj, N. J., Santosa, K. B., Newton, P., Liu, Z., Hunter, D. A., Mackinnon, S. E., Sakiyama-Elbert, S. E., & Johnson, P. J. (2011). A systematic evaluation of Schwann cell injection into acellular cold-preserved nerve grafts. Journal of Neuroscience Methods, 197(2), 209-215. https://doi.org/10.1016/j.jneumeth.2011.02.015