1.Osteogenetic ability of mouse spermatogonial stem cells cultured in vitro
Hongmei HU ; Wei LI ; Xinming SUN ; Fen JIANG ; Ganjun YANG
Chinese Journal of Tissue Engineering Research 2014;(23):3702-3706
BACKGROUND:Spermatogonial stem cells are a kind of adult stem cells, which have self-renewal and differentiation potential, and can be differentiated into specific cells in vitro, suggesting that the spermatogonial stem cells may be possibly differentiated into osteoblasts. But the related research has not been reported. OBJECTIVE:To observe the biological characterization and osteogenic process of mouse spermatogonial stem cells cultured in vitro. METHODS:Spermatogonial stem cells were obtained from the testicle of mice aged 15-20 days, and were cultured on the feeder layer from bone marrow stroma cells in vitro. When cultured for 3 days, the cells were cultured in the conditioned medium (experimental group) and basic medium (control group). The cells proliferation capability and osteogenic property were examined by phase-contrast microscope, alkaline phosphatase activity and type I col agen immunofluorescence staining. RESULTS AND CONCLUSION:Spermatogonial stem cells proliferated faster in the experiment group than in the control group. cells grew rapidly in colony-like shape in the conditioned medium at 3-6 days, the three-dimensional feeling enhanced, cellmass and clusters continued to increase in size, the extracellular matrix was increased in number and the cytoplasmic bridge was not obvious. After culture for 15 days, cells in the two groups were positive for alkaline phosphatase staining that the cytoplasmic membrane was dyed black. Under the fluorescent microscope, green fluorescence was visible in the experimental group, suggesting the cells in the experimental group was positive for type I col agen, but negative in the control group, which is similar with the biological characteristics of osteoblasts. These findings indicate that spermatogonial stem cells possess the osteogenic capability under induction conditions, which are expected to provide seed cells for bone tissue engineering.
2.The influence of MCP-1/CCR2 axis mediated NF-κB signal pathway on the migration of dental pulp stem cells steamulated by inflamation factor
Wei LI ; Jian'an PENG ; Ganjun YANG ; Hongmei HU
Journal of Practical Stomatology 2024;40(5):638-646
Objective:To explore the molecular mechanism of NF-κB signaling pathway mediated by MCP-1/CCR2 axis in the migra-tion of dental pulp stem cells(DPSCs)stemulated by inflamation factor.Methods:DPSCs of SD rats were in vitro cultured,and identified by flow cytometry.The inflammation model of DPSCs was established by LPS treatment,Western blot and Q-PCR were used to detect the expression of MCP-1 and CCR2 in DPSCs of the groups.The inflamatory DPSCs were treated by NF-κB pathway activator TNF-α and NF-κB pathway inhibitor BMS-345541 respectively,the activation of P65 in inflammatory DPSCs was detected by immuno-fluorescence.Scratch experiment was used to detect the migration ability of DPSCs.Results:Higher expression of MCP-1 and CCR2 genes was observed in LPS-induced DPSCs than in the controls,TNF-α treated DPSCs showed higher expression of MCP-1 and CCR2 genes than the LPS tread and the NF-κB pathway inhibitor BMS-345541 treated cells.TNF-α treated DPSCs showed higher scratch healing effect than LPS treated and BMS-345541 treated cells.Conclusion:The expression of MCP-1 and CCR2 genes in DPSCs is higher in inflammatory state,and the activation of NF-κB signaling pathway can enhance this expression,which can ultimately promote the migration ability of DPSCs in inflammatory state.
3.Application of single-direction gastric mobilization under 3D-laparoscopy in minimally invasive esophagectomy for the treatment of esophageal cancer
Wanli JIANG ; Ganjun KANG ; Xin WANG ; Hengya SONG ; Sen YANG ; Jie HUANG ; Songping XIE
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2021;28(01):31-34
Objective To evaluate the safety, feasibility and short-term outcomes of single-direction gastric mobilization under 3D-laparoscopy in minimally invasive esophagectomy for the treatment of esophageal cancer. Methods From February 2018 to December 2019, 118 consecutive patients who underwent minimally invasive McKeown esophagectomy for esophageal squamous cell carcinoma in our hospital were included. There were 94 males and 24 females with an average age of 53.7 (41–77) years. They were divided into two groups based on the methods of gastric mobilization: a traditional dissociation (TD) group (n=55) and a single-direction mobilization (MD) group (n=63). The clinical data of the two groups were compared. Results Enbloc resection and a negative resection margin were obtained in all patients. There was no postoperative mortality or incision complication. The rate of postoperative complications was 22.9%. There was no significant difference in the spleen injury, gastric injury, conversion to open surgery, abdominal reoperation as well as cervical anastomotic leakage between the two groups (P>0.05). It took significantly less time in the MD group compared with the TD group (P<0.05). There was an obvious statistical difference in the incidence of gastric mobilization related complications between the MD group (1.6%, 1/63) and TD group (12.7%, 7/55, P<0.05). Conclusion Application of single-direction gastric mobilization under 3D-laparoscopy in minimally invasive esophagectomy for the treatment of esophageal cancer is safe and easy to perform with a satisfactory short-term outcome.