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3D Hydrogel Scaffolds for Articular Chondrocyte Culture and Cartilage Generation
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Development of an efficient vitrification method for chondrocyte sheets for clinical application.

Asuka Hayashi1, Miki Maehara2, Ayuko Uchikura1

  • 1Laboratory of Medical Bioengineering, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama, Kawasaki 214-8571, Japan.

Regenerative Therapy
|May 22, 2020
PubMed
Summary

A new circulating vitrification bag and storage box simplify chondrocyte sheet cryopreservation. This method maintains cell viability and cartilage component integrity, advancing regenerative therapy for osteoarthritis.

Keywords:
Cell sheetChondrocyteCryopreservationDMSO, dimethyl sulfoxideEG, ethylene glycolFBS, fetal bovine serumLN, liquid nitrogenOsteoarthritisPBS, phosphate buffered salineVitrification

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Area of Science:

  • Biomedical Engineering
  • Regenerative Medicine
  • Cryobiology

Background:

  • Chondrocyte sheet therapy shows promise for osteoarthritis treatment.
  • Effective cryopreservation is crucial for advancing clinical applications of chondrocyte sheets.
  • Previous vitrification methods were too complex for routine clinical use.

Purpose of the Study:

  • To develop a prototype method for vitrifying chondrocyte sheets suitable for clinical practice.
  • To create a simpler and more feasible cryopreservation technique for chondrocyte sheets.

Main Methods:

  • Development of a "circulating vitrification bag" for efficient and sanitary processing of cell sheets.
  • Invention of a "vitrification storage box" for vitrification, long-term preservation, and transportation.
  • Vitrification of rabbit chondrocyte sheets using the developed devices and assessment of structural integrity and cell viability.

Main Results:

  • The circulating vitrification bag method maintained chondrocyte sheet structural integrity.
  • Cell survival rates after vitrification were comparable to non-vitrified samples (91.0 ± 2.9% vs. 90.0 ± 3.0%).
  • Proteoglycan and type II collagen distribution remained dense and even post-vitrification, and long-term storage in the vitrification storage box preserved cell viability (81.2 ± 1.0% vs. 84.3 ± 1.8%).

Conclusions:

  • The circulating vitrification bag method is effective for clinical application of vitrified chondrocyte sheets.
  • The vitrification storage box facilitates long-term preservation and transportation of vitrified cell sheets.
  • These advancements enhance the feasibility of clinical application for cryopreserved chondrocyte sheets in regenerative therapy.