Establishment of Banking System for Allogeneic Cultured Dermal Substitute

Abstract

Allogeneic cultured dermal substitute (CDS) was prepared by culturing fibroblasts on a two-layered spongy matrix of hyaluronic acid (HA) and atelo-collagen (Col). Allogeneic CDS can be cryopreserved and transported to other hospitals in a frozen state. Vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), platelet derived growth factor (PDGF)-AA, transforming growth factor (TGF)-beta1, keratinocytes growth factor (KGF), interleukin (IL)-6 and IL-8 were contained in the culture medium which was used in preparing CDS over a cultivation period of one week (fresh CDS culture medium sample). After thawing a cryopreserved CDS, the CDS was recultured in a culture medium for one week. VEGF, bFGF, HGF, TGF-beta1 and IL-8 were contained in the culture medium which was used in reculturing CDS for one week (cryopreserved CDS culture medium sample), although some cytokines were detected at a lower level than those before freezing. This finding suggests that the cryopreserved CDS retains its ability to release these cytokines. Clinical research on allogeneic CDS, which was newly developed at the R & D Center for Artificial Skin of Kitasato University, has been carried out in medical centers across Japan with the support of the Millennium Project of the Ministry of Health, Labor and Welfare. It was demonstrated that the allogeneic CDS functions as an excellent cell therapy for intractable skin ulcers as well as burn injuries. The spongy matrix itself, as well as the cytokines released from the allogeneic CDS, seemed to be beneficial for the treatment of intractable skin defect.

Effects of Freezing on Functionality and Physicochemical Properties of A 3D-Human Skin Model

“…Different methods were used for epidermal substitute cryopreservation, such as the use of dimethyl sulfoxide [6,31,32] and trehalose as cryoprotectants [6]. Dimethyl sulfoxide has also been tested on dermal substitutes [33,34]. Cells' viability was measured to evaluate the impact of these cryoprotectants.…”

Review Paper: Burn Coverage Technologies: Current Concepts and Future Directions

“…Allogeneic CDSs can be prepared by culturing fibroblasts on a specially designed scaffold, such as two-layered spongy matrix of hyaluronic acid and atelocollagen [41]. A biodegradable salt-leached porous gelatinous scaffold is also appropriate to seed cultured fibroblasts.…”

“…After 14 days of culturing, the fibroblasts exhibit a good affinity to and proliferation on the gelatin scaffolds without showing any signs of biodegradation [40]. CDS can be cryopreserved and transported to other hospitals in a frozen state retaining its ability to release essential cytokines for wound healing particularly vascular endothelial growth factor (VEGF) [41]. Taking into account the manufacturing cost, coupled with the potency of VEGF release, a two-layered sponge of HA and Col with a weight ratio of 5/2 is very promising for commercial application [41].…”

<i>In Vitro</i> Analysis of VEGF and HGF Production by Fibroblast in Cultured Dermal Substitute Combined with EGF-Incorporating Top Dressing

“…The wound dressing is composed of hyaluronic acid (HA) and collagen (Col) spongy sheet containing epidermal growth factor (EGF) [1] [2]. The cultured dermal substitute (CDS) is composed of HA and Col spongy sheet containing human fibroblasts [3] [4]. Fibroblasts in CDS are capable of releasing various types of cytokines and extracellular matrix, which are necessary for wound healing.…”