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Autologous cell sheet engineering: an animal product‐free approach (1180.23)
Author(s) -
Wood Andrew,
Sota Hiroyuki,
OlivaVilana Joan,
BardagGorce Fawzia,
Thropay Jacquelyn,
Pan Derek,
Niihara Yutaka
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1180.23
Subject(s) - tissue engineering , staining , ex vivo , cell , in vivo , epithelium , basal (medicine) , cell culture , microbiology and biotechnology , chemistry , in vitro , biomedical engineering , biology , pathology , medicine , biochemistry , genetics , insulin
While ex vivo expanded autologous cell sheet shows promise in replacing impaired corneal epitheliums; the engineering issues of using xenogeneic products during cell expansion remain to be addressed. To eliminate animal products, this study explores the use of two chemically defined media used sequentially for expansion and differentiation, where submerged and air‐exposure techniques are used respectively. The cells in the xenogeneic product‐free condition expanded well and reached confluency however only a single layer of basal cells was created. To resolve this issue, we found that the addition of 1 mM Ca2+ helps induce differentiation. However, air‐exposure could cause the apical surface to become dry and keratinized. To overcome this downside, a few drops of PBS as a moistener, were applied to the surface. The modified culture with this preventive technique successfully yielded non‐keratinized, multi‐stratified cell sheets that are morphologically similar to cell sheets engineered with the conventional method. These observations were confirmed by H&E staining. Moreover, the sheet maintained the normal phenotypes; the basal cells stained positive for ΔN‐p63, Ki67 and PCNA, while the apical cells were stained for K3 and K13. The chemically defined non‐xenogeneic media showed promise in becoming a safe alternative for the epithelium production. Supported by Emmaus Life Sciences, Inc. and CellSeed, Inc.