Development of frozen-thawed allogeneic fibroblast sheets for esophageal anastomotic leakage

Esophageal anastomotic leakage is a severe postoperative complication for which effective preventive methods have not yet been established. Previous studies have shown that allogeneic multilayered fibroblast sheets developed in our laboratory secrete various growth factors that promote wound healing. However, for clinical application, a method for long-term preservation is essential. In this study, we focused on cryopreservation using a 3-dimensional freezer to establish a method for long-term preservation. The aim of this study was to evaluate the efficacy of cryopreserved allogeneic multilayered fibroblast sheets in preventing esophageal anastomotic leakage.

Cryopreserved allogeneic fibroblast sheets were prepared from rat oral mucosa. A rat esophageal anastomosis model was created, and 2 transplanted cell sheets were used. The incidence of anastomotic leakage, burst pressure, histologic findings, and collagen content was evaluated postoperatively.

The multilayered fibroblast sheets group exhibited a significantly lower incidence of anastomotic leakage (control: 11/18, 61% vs sheet: 5/18, 28%; P = .02). The burst pressure was significantly greater in the sheet group (control: 78 ± 26 mm Hg vs sheet: 124 ± 30 mm Hg; P < .01), as was the collagen content (control: 0.09 [0.05-0.13] μg vs sheet: 0.15 [0.11-0.17] μg; P = .04). The sheet group exhibited a significant increase in the collagen fiber area (control: 0.41 [0.37-0.44] mm²/mm² vs sheet: 0.53 [0.49-0.58] mm²/mm²; P < .01).

The transplantation of cryopreserved allogeneic multilayered fibroblast sheets improved the incidence of anastomotic leakage and burst pressure, suggesting their potential role in preventing esophageal anastomotic leakage.