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BACKGROUND: Induced pluripotent stem cells (iPSCs) are a special type of cells with self-renewal and multi-differentiation potential, which can differentiate into intestinal organoids under certain conditions. OBJECTIVE: To explore whether iPSCs can differentiate into intestinal organoids under specific conditions in vitro.METHODS: iPSCs from B6J mice were recovered and cultured for 3 days until clone units covered about 80％ of the culture dish, and then the cells were cultured in the medium containing Activin A for 3 days until the deterministic endoderm formed. Further, the culture medium was replaced by the medium with fibroblast growth factor 4 and Wnt3A for 4 days to differentiate into the spheroids with CDX2+. After that, spheroids were collected and mixed with Matrigel,and then the mixture was dropped into the 4-well plate and cultured with Rspondin1, Noggin, epidermal growth factor, B27 and other growth factors to differentiate into intestinal organoids. Cell morphology was observed, FoxA2 and Sox17 expresson in the deterministic endoderm was detected, and CDX2, Sox9, CGA, MMP7 were measured.RESULTS AND CONCLUSION: iPSCs were cultured with Activin A for 3 days with higher cell fusion, initial differentiation and FoxA2/Sox17 expression (P < 0.05) than those of non-induced iPSCs. Spheroids began to appear at the 3rd day after culture with fibroblast growth factor 4 and WNT3A, and formed a lot at the 4th day. And CDX2 expression in spheroids was significantly increased compared with that in the deterministic endoderm (P < 0.05). Organoids gradually formed after 3 days culture, which contained all cell types of intestinal organoids, and expressions of specific markers, Sox9, CGA, MMP7, were significantly higher than those in spheroids (P < 0.05). To conclude, iPSCs can be induced to differentiate into intestinal organoids in three-dimensional niche in vitro.
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Objective To investigate effective treatment modalities and the related factors influencing prognosis of patients with gallbladder cancer.Methods The clinical data of 76 gallbladder carcinoma patients admitted to the Department of General Surgery,PLA Nanjing General Hospital from January 2005 to October 2015 were analyzed retrospectively.Follow-up was carried out via telephone or outpatient service until January 2016.Cox regression and Kaplan-Meier models were performed for survival analysis.Results 69 patients were treated with surgery and/or postoperative adjuvant chemotherapy.The remaining 7 patients with liver or distant metastases who did not undergo surgery received chemotherapy.24 patients died from cancer relapse,37 patients died from disease progression after giving up treatment,and 7 patients were lost to follow-up.The remaining 8 patients were still alive at the time of follow-up.The depth of cancer invasion (HR =2.736),the type surgical procedure (HR =2.207),and adjuvant chemotherapy (HR =0.603) were significant impact factors of survival for GBC patients.Adjuvant chemotherapy was a protective factor.The average survival in the chemotherapy-naive group was (10.6 ± 1.9) months,the single chemotherapy group (18.5 ± 2.8) months,and the combined chemotherapy group (26.9 ± 6.4) months.There were no significant differences among these groups.Conclusions The depth of cancer invasion,types of surgical procedure particularly radical cholecystectomy,and adjuvant chemotherapy were significant factors of survival in patients with GBC.Radical cholecystectomy combined with arterial and intravenous chemotherapy using gemcitabine and oxaliplatin showed benefits in survival in GBC patients.
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Sepsis is the leading cause of mortality in critically ill patients.Studies indicate that immune suppression in sepsis is more often associated with poor outcome.Dendritic cells may contribute largely to the development of immune suppression during sepsis.This article reviews the emerging data indicating the key role of dendritic cells in sepsis induced immune suppression.A deeper insight into the dendritic cell changes during sepsis may provide a powerful weapon against sepsis.