Fabrication of transplantable corneal epithelial and oral mucosal epithelial cell sheets using a novel temperature-responsive closed culture device

Abstract

Temperature-responsive culture surfaces make it possible to harvest transplantable carrier-free cell sheets. Here, we applied temperature-responsive polymer for polycarbonate surfaces with previously developed closed culture devices for an automated culture system in order to fabricate transplantable stratified epithelial cell sheets. Histological and immunohistochemical analyses and colony-forming assays revealed that corneal epithelial and oral mucosal epithelial cell sheets could be harvested with the temperature-responsive closed culture devices. The results were similar to those obtained using temperature-responsive culture inserts. These results indicate that the novel temperature-responsive closed culture device is useful for fabricating transplantable stratified epithelial cell sheets.

Keratinocyte sheets prepared with temperature-responsive dishes show enhanced survival after in vivo grafting on acellular dermal matrices in a rat model of staged bi-layered skin reconstruction

“…As rapid hydration of PIPAAm occurs, a separating layer forms and cells can be collected as a single contiguous cell sheet [22]. TR techniques have been successfully adopted to develop cell sheets for in vivo repair of corneal defects [25], [26], [27], esophageal mucosa [28], oral and nasal mucosal lining [29], ear canal mucosa [30], [31], cartilage [32], and endometrium [33], [34], among others. Besides positive outcomes observed in animal models, initial studies in patients are also promising [35].…”

A novel application of electrospun silk fibroin/poly(l-lactic acid-co-ε-caprolactone) scaffolds for conjunctiva reconstruction

“…H&E staining was performed on both in vitro and in vivo samples as previously described. 27 Briefly, in vitro and in vivo cell-seeded scaffolds were embedded in Tissue-Tek OCT compound (Sakura Seiki, Tokyo, Japan), quickly frozen and cut into 10 mm-thick sections. After being dried for 30 min at room temperature, the sections were fixed with 4% w/v PFA at room temperature for 15 min, then washed three times with distilled-deionized water (DDW) and stained with H&E. Microphotographs were taken with a light microscope (Olympus BX51, Japan).…”

Conjunctival reconstruction via enrichment of human conjunctival epithelial stem cells by p75 through the NGF-p75-SALL2 signaling axis

“…Histomorphology was detected by H&E staining as previously described. 43,44 Briefly, tissue samples were cut into 8 μm sections and stained with H&E. Immunohistochemical staining also followed the previously described method. Briefly, the sections were incubated 3 | RESULTS…”