BACKGROUND: Recent studies have shown that induced pluripotent stem cells (iPSCs) could be differentiated into mesenchymal stem cells (MSCs) with notable advantages over iPSCs per se. In order to promote the application of iPSC-MSCs for osteoregenerative medicine, the present study aimed to assess the ability of murine iPSC-MSCs to differentiate into osteoblast phenotype. METHODS: Osteogenic differentiation medium, blending mouse osteoblast-conditioned medium (CM) with basic medium (BM) at ratio 3:7, 5:5 and 7:3, were administered to iPSC-MSCs, respectively. After 14 days, differentiation was evaluated by lineage-specific morphology, histological stain, quantitative reverse transcription-polymerase chain reaction and immunostaining. RESULTS: The osteogenesis-related genes, alp, runx2, col1 and ocn expressions suggest that culture medium consisting of CM:BM at the ratio of 3:7 enhanced the osteogenic differentiation more than other concentrations that were tested. In addition, the alkaline phosphatase activity and osteogenic marker Runx2 expression demonstrate that the combination of CM and BM significantly enhanced the osteogenic differentiation of iPSC-MSCs. CONCLUSION: In summary, this study has shown that osteoblast-derived CM can dramatically enhance osteogenic differentiation of iPSC-MSCs toward osteoblasts. Results from this work will contribute to optimize the osteogenic induction conditions of iPSC-MSCs and will assist in the potential application of iPSC-MSCs for bone tissue engineering.
Alkaline Phosphatase ; Animals ; Bone and Bones ; Culture Media, Conditioned ; Induced Pluripotent Stem Cells ; Mesenchymal Stromal Cells ; Mice ; Osteoblasts ; Phenotype

Objective To observe the metabolomic changes of amniotic fluid in control group and administration group,and to explore the toxicity of aqueous extract of rhubarb on reproduction and embryonic development of pregnant rats.Methods Pregnant rats in teratogenic sensitive period were given rhubarb aqueous extract by gavage for 10 days.The toxicity of rhubarb to maternal rats and the abnormal conditions of dead fetus and absorbed fetus were observed.The amniotic fluid samples were detected by liquid chromatography-mass spectrometry(LC-MS),the amniotic fluid metabolic profiles were compared by principal component analysis(PCA)and partial least squares(OPLS-DA),and the differential lipids were analyzed by metaboanalyst 5.0.Results Rhubarb administration in the teratogenic sensitive period can significantly reduce the number of live fetuses,and lead to adverse phenomena such as absorption of fetuses and premature death of fetuses.The preliminary results of lipomics showed that rhubarb could cause the metabolic disorder of amniotic fluid in pregnant rats,and there were metabolic abnormalities in lipids in amniotic fluid such as PI,PC and LPC.Conclusion Under the equivalent dose of the maximum dose recommended by the clinic,the aqueous extract of rhubarb has certain reproductive and embryonic toxicity to rats;Rhubarb may affect the normal development of rat embryos by causing the disorder of lipid metabolism in amniotic fluid.