Catharmus tinctorius volatile oil promote the migration of mesenchymal stem cells via ROCK2/Myosin light chain signaling / 中国天然药物

MSC transplantation has been explored as a new clinical approach to stem cell-based therapies for bone diseases in regenerative medicine due to their osteogenic capability. However, only a small population of implanted MSC could successfully reach the injured areas. Therefore, enhancing MSC migration could be a beneficial strategy to improve the therapeutic potential of cell transplantation. Catharmus tinctorius volatile oil (CTVO) was found to facilitate MSC migration. Further exploration of the underlying molecular mechanism participating in the pro-migratory ability may provide a novel strategy to improve MSC transplantation efficacy. This study indicated that CTVO promotes MSC migration through enhancing ROCK2 mRNA and protein expressions. MSC migration induced by CTVO was blunted by ROCK2 inhibitor, which also decreased myosin light chain (MLC) phosphorylation. Meanwhile, the siRNA for ROCK2 inhibited the effect of CTVO on MSC migration ability and attenuated MLC phosphorylation, suggesting that CTVO may promote BMSC migration via the ROCK2/MLC signaling. Taken together, this study indicates that C. tinctorius volatile oil could enhance MSC migration via ROCK2/MLC signaling in vitro. C. tinctorius volatile oil-targeted therapy could be a beneficial strategy to improve the therapeutic potential of cell transplantation for bone diseases in regenerative medicine.

Effects of human aorta-gonad-mesonephros region stromal cells on inducing differentiation of murine embryonic stem cells into hematopoietic stem/progenitor cells in vitro / 中国实验血液学杂志

This study was aimed to investigate the effect of human aorta-gonad-mesonephros (AGM) region stromal cells on differentiation of murine embryonic stem cells (ESC) into hematopoietic stem cells (HSC) in vitro and to clarify their effect mechanism. E14 murine ESC were induced into embryo body (EB) firstly. Then the EB cells were further co-cultured with the stromal cells from human AGM region, fetal liver (FL) or bone marrow (BM) in Transwell non-contact system. According to the different culture methods, the EB cells were divided into 6 groups including EB control group, AGM group, FL group, BM group, AGM + FL group and AGM + BM group. The induced cells derived from EB were collected for Sca-1(+)/c-Kit(+) cells analysis by flow cytometry and colony forming unit (CFU) assay. The results showed that Sca-1(+)/c-Kit(+) cell proportion of EB cells significantly increased after being induced by different stromal cells (p < 0.01). The AGM + FL group had most Sca-1(+)/c-Kit(+) cells for the positive cell proportion reached (21.96 ± 2.54) % (p < 0.01). The Sca-1(+)/c-Kit(+) cell proportion of AGM + BM group was much high than that of BM group too (p < 0.01). The EB control group showed CFU amount less than any other groups, while the CFU amount of AGM + FL, AGM + BM groups were higher, especially in the AGM + FL group (p < 0.01). It is concluded that the human AGM region stromal cells may help to maintain certain number of primitive HSC with high proliferation potential. Human AGM region, FL or BM stromal cells, applied in sequential orders, can significantly expand in vitro the primitive hematopoietic stem/progenitor cells derived from ESC.

Sequentially inducting murine embryonic stem cells into hematopoietic stem cells in vitro by hematopoietic development procedure for reconstitution of hematopoiesis in vivo / 中国实验血液学杂志

This study was purposed to directly induce murine embryonic stem cells (ESC) into hematopoietic stem cells (HSC) by simulating the spatial and temporal hematopoietic microenvironment changes in embryonic development, and to investigate the function of in vivo hematopoietic reconstitution of these HSC. E14 ESC were induced into embryoid body (EB) firstly. Then the cells from EB were further co-cultured with human aorta-gonad-mesonephros (AGM) region, fetal liver (FL) and bone marrow (BM) stromal cells in Transwell non-contact system in sequential orders. After 6 days of each co-cultured stage, the induced cells derived from EB were collected to analyze the Sca-1(+)c-Kit(+) cells by flow cytometry, check teratoma formation and transplant to BALB/C female mice exposed to lethal dose (60)Co γ-ray. The recipient mice were divided randomly into 5 groups: AGM, AGM + FL, AGM + FL + BM, irradiation control and normal control groups. The survival rates, hematopoietic reconstitution and engraftment of donor cells in the different groups were monitored. The results showed that Sca-1(+)c-Kit(+) cell level in EB cells co-cultured with human AGM region and FL stromal cells reached to peak value (21.96 ± 2.54)%. Teratomas could be found in NOD-SCID mice after subcutaneous injection of EB cells co-cultured with human AGM region stromal cells, while there was no teratoma in the mice after subcutaneous injection of EB cells induced by human AGM region and FL stromal cells. The recipients in AGM group and irradiation control group all died. The survival rate was 77.8% in AGM+FL group, and 66.7% in AGM+FL+BM group. The peripheral blood cell count was near normal at day 21 after transplantation, and Sry gene copies from donor could be detected in recipient mice of these two groups. It is concluded that sequentially inductive system with feeder cells from human AGM region, fetal liver and bone marrow simulating embryonic defined hematopoiesis procedures can enhance the directed differentiation of ESC into HSC which can safely reconstitute hematopoiesis in vivo.