Abstract
Temporary storage of nasal tissues and nasal cell sheets, which entails transportation between hospitals and cell culture facilities, is an important issue in regenerative medicine. Herein, we investigated the preservation of chilled and frozen nasal tissues and expiry dates of ready-to-use nasal cell sheets. Although the cell number in preserved tissues was lower than that in fresh tissue, nasal cell sheets could be fabricated from tissues that had been refrigerated for 5 days and frozen–thawed over 5 days. Moreover, the nasal mucosal cell sheets were preserved in a non-hazardous buffer. The cell number, viability, and structure were not maintained in saline containing E-cadherin for 2 days; however, these were maintained in Hank’s balanced salt solution for 2 days, but not for 5 days. To assess the proliferation capacity of cells in the stored cell sheets, we performed cell sheet grafting assays in vitro. Cell sheets stored in Hank’s balanced salt solution for 2 days adhered to collagen gel and expanded normally. Our results show that nasal tissues can be stored temporarily in refrigerators or deep freezers, and Hank’s balanced salt solution can be used for preservation of ready-to-use cell sheets for a few days.
Highlights
Intractable otitis media, cholesteatoma, and adhesive otitis media were successfully treated using autologous nasal mucosal cell sheet as a regenerative medicine in a clinical study (Yamamoto et al, 2017)
We previously showed that both nasal tissue and nasal mucosal cell sheets can be transported for 3 h without a decrease in quality (Kasai et al, 2019)
To detect the proliferation activity of the cells in fresh tissue stored at 4◦C for 5 days and at −80◦C for 19–70 days, the expression of proliferation marker, Ki-67, and apoptosis marker, poly (ADP-ribose) polymerase (PARP), was examined by immunohistological analysis
Summary
Intractable otitis media, cholesteatoma, and adhesive otitis media were successfully treated using autologous nasal mucosal cell sheet as a regenerative medicine in a clinical study (Yamamoto et al, 2017). Cells from cryopreserved umbilical cord or adipose tissues have the ability to grow and differentiate (Shimazu et al, 2015; Arutyunyan et al, 2018; Zanata et al, 2018); suspended cells, native tissues can be stored in a deep freezer. In this context, we hypothesized that the Abbreviations: CFE, colony-forming efficiency; HBSS, Hank’s balanced salt solution; HE staining, hematoxylin-eosin staining; KCM, keratinocyte culture medium; PARP, poly (ADP-ribose) polymerase
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