Applicable possibility of autologous osteochondral regenerative medicine using umbilical cord-derived mesenchymal stromal cells of cleft palate patient

Abstract

Background & Aim Cleft palate is a major obstacle to constraining ADL for the children. In this study, to regenerate and compensate the defected palate by autologous umbilical cord-derived mesenchymal stromal cells (UC-MSC) of cleft palate patient, we studied the osteochondrogenic differentiation potency of UC-MSC. Methods, Results & Conclusion Materials/Methods UC-MSC derived from a patient who had cleft palate (CLP-UC-MSC) at the time of birth was obtained from the umbilical cord blood and cord bank of the Institute of Medical Science, The University of Tokyo (IMSUT CORD). We also obtained MSC derived from two healthy children (H-UC-MSC) as control and compared them. Banking and the experiments of this study were approved by IRB of IMSUT. Result The proliferation and surface markers of CLP-UC-MSC (CD105 + D73 + CD90 + HLA class I + CD45-HLA-DR-) were equivalent to H-UC-MSC. Expanded CLP-UC-MSC at passage 4 (P4) showed normal karyotype. Osteogenic differentiation using P4 cells was performed by seeding 12-well plates in 1 ml of the osteogenic differentiation medium, replacing the medium every 5 days, and culturing for 30 days. In Alizarin Red S staining, deposited Ca was confirmed in differentiated CLP-UC-MSC as well as those in the H-UC-MSC. Chondrogenic differentiation potency using P4 cells was evaluated by a pellet method in 15ml centrifuge tube made of PP, adding chondrogenic differentiation medium. After 24 days of culture, intracellular mucoitinsulfuric acid and chondroitin sulfate stained by Toluidine Blue Solution was observed. Discussion In this study, we demonstrated that CLP-UC-MSC showed the equivalent osteochondrogenic differentiation potential to H-UC-MSC, although only one case of cleft palate was examined. It was reported previously that UC-MSC showed less osteogenic differentiation ability compared to bone marrow-derived MSC, but we succeeded to induce the osteogenic differentiation potency, suggesting that it could be applied to osteochondral regeneration using autologous CLP-UC-MSC and H-UC-MSC.