Amniotic membrane of human placenta is a source of abundant mesenchymal stem cell (hAMSC) which makes it a potential source of allogeneic multipotent cell for bone healing. However, much has to be explored about its i...Amniotic membrane of human placenta is a source of abundant mesenchymal stem cell (hAMSC) which makes it a potential source of allogeneic multipotent cell for bone healing. However, much has to be explored about its isolation procedure and the osteogenic differentiation potential. The aims of this study are to establish the procurement procedure of human amniotic membrane, the isolation and culture of hAMSC, the MSC phenotypic characterization, and the in vitro osteogenic differentiation of hAMSC. Results of the study are as follows. The quality of human amniotic membrane would be best if procured from Caesarean operation under highly aseptic condition to avoid fungal and bacterial contamination on the culture. Isolation procedure using modified Soncini protocol yielded large amount of MSC with high proliferative capacity in culture medium. Characterization of hAMSC showed that the majority of the target cells exhibited specific MSC markers (CD105 and CD90) with a small number of these cells expressing CD45, the marker of hematopoeitic cells. The in vitro osteogenic differentiation of hAMSC followed by Alizarin Red staining showed that osteoblastic differentiation was detected in a significantly high number of cells. This study concludes that hAMSCs isolated from human amniotic membrane have the capacity for in vitro osteogenesis which makes them be one of the potential allogeneic stem cells for application in maxillofacial bone reconstruction.展开更多
The human umbilical cord is a source of numerous Mesenchymal Stem Cells (MSCs), making it as a potential source of allogeneic multipotent cell for bone tissue engineering. The aims of this study were to find: 1) Human...The human umbilical cord is a source of numerous Mesenchymal Stem Cells (MSCs), making it as a potential source of allogeneic multipotent cell for bone tissue engineering. The aims of this study were to find: 1) Human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) phenotypic characterization, 2) The in-vitro osteogenic differentiation potential of hUCMSCs, 3) The cytotoxicity of gelatin solvent to hUCMSCs using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. As a result, through characterization of hUCMSCs, the majority of target cells expressed specific MSCs markers, Cellular Differentiation (CD)73, smaller number of subpopulation expressed CD90 with only minimal subpopulation expressed CD105 and all negative MSCs markers. Osteoblastic differentiation was found in a significantly high number of cells when in vitro osteogenic differentiation of hUCMSCs with Alizarin Red staining was done. Biocompatibility analysis using the MTT assay showed that gelatin solvent and Alpha modification of minimum essential medium Eagle (α-MEM) was non-toxic for hUCMSCs in vitro. The study concluded that hUCMSCs isolated from human umbilical cord was capable of undergoing in vitro osteogenesis, indicating its potential as allogeneic stem cells for clinical application in bone tissue engineering.展开更多
文摘Amniotic membrane of human placenta is a source of abundant mesenchymal stem cell (hAMSC) which makes it a potential source of allogeneic multipotent cell for bone healing. However, much has to be explored about its isolation procedure and the osteogenic differentiation potential. The aims of this study are to establish the procurement procedure of human amniotic membrane, the isolation and culture of hAMSC, the MSC phenotypic characterization, and the in vitro osteogenic differentiation of hAMSC. Results of the study are as follows. The quality of human amniotic membrane would be best if procured from Caesarean operation under highly aseptic condition to avoid fungal and bacterial contamination on the culture. Isolation procedure using modified Soncini protocol yielded large amount of MSC with high proliferative capacity in culture medium. Characterization of hAMSC showed that the majority of the target cells exhibited specific MSC markers (CD105 and CD90) with a small number of these cells expressing CD45, the marker of hematopoeitic cells. The in vitro osteogenic differentiation of hAMSC followed by Alizarin Red staining showed that osteoblastic differentiation was detected in a significantly high number of cells. This study concludes that hAMSCs isolated from human amniotic membrane have the capacity for in vitro osteogenesis which makes them be one of the potential allogeneic stem cells for application in maxillofacial bone reconstruction.
文摘The human umbilical cord is a source of numerous Mesenchymal Stem Cells (MSCs), making it as a potential source of allogeneic multipotent cell for bone tissue engineering. The aims of this study were to find: 1) Human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) phenotypic characterization, 2) The in-vitro osteogenic differentiation potential of hUCMSCs, 3) The cytotoxicity of gelatin solvent to hUCMSCs using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. As a result, through characterization of hUCMSCs, the majority of target cells expressed specific MSCs markers, Cellular Differentiation (CD)73, smaller number of subpopulation expressed CD90 with only minimal subpopulation expressed CD105 and all negative MSCs markers. Osteoblastic differentiation was found in a significantly high number of cells when in vitro osteogenic differentiation of hUCMSCs with Alizarin Red staining was done. Biocompatibility analysis using the MTT assay showed that gelatin solvent and Alpha modification of minimum essential medium Eagle (α-MEM) was non-toxic for hUCMSCs in vitro. The study concluded that hUCMSCs isolated from human umbilical cord was capable of undergoing in vitro osteogenesis, indicating its potential as allogeneic stem cells for clinical application in bone tissue engineering.
