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Our study shows that conditioned medium from renal tubular epithelial cells provides a convenient source of inductive signals to initiate differentiation of ASC towards epithelial lineage. We deduce that these interactions may play an important role during renal repair mechanisms.

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The versatile differentiation potential of adipose-derived stem cells (ASC) into cells of mesodermal, entodermal, and ectodermal origin places these cells at the forefront of cell-based therapies and cell transplantation. Epithelial differentiation of ASC may either be initiated by direct cell-cell or cell-matrix contacts, by chemical factors like retinoic acid, or via secreted cellular factors like cytokines, interleukins, or growth factors included in conditioned media.This protocol describes methods to induce the in vitro differentiation of ASC from human adipose tissue into the epithelial lineage, and describes the methods used to verify this induced differentiation. We present two differentiation protocols based on either retinoic acid or conditioned medium of cultured epithelial cells.

SUMMARYClinical relevance of ELISA-and single-antigen bead assay (SAB)-detected pretransplant HLA antibodies (SAB-HLA-Ab) for kidney graft survival was evaluated retrospectively in 197 patients transplanted between 2002 and 2009 at the University Clinic Frankfurt. Having adjusted for retransplantation and delayed graft function, a significantly increased risk for death-censored graft loss was found in patients with pretransplant SAB-HLA-Ab [HR: 4.46; 95% confidence interval (CI): 1.47-13.48; P = 0.008]. The risk for increased graft loss was also significant in patients with pretransplant SAB-HLA-Ab but without SAB-detected donor-specific Ab (SAB-DSA) (HR: 4.91; 95% CI of 1.43-16.991; P = 0.012). ELISA was not sufficient to identify pretransplant immunized patients with an increased risk for graft loss. In immunized patients, graft loss was predominantly present in patients who received transplants with a mismatch on the HLA-DR locus. In conclusion, even if our study is limited due to small sample size, the results show an increased risk for long-term graft loss in patients with pretransplant SAB-HLA, even in the absence of DSA. SAB-HLA-Ab-positive patients, being negative in ELISA or CDC assay, might profit from a well-HLA-DR-matched graft and intensified immunosuppression. Transplant International 2016; 29: 988-998