AB194. Adipose derived stem cell differentiated towards urothelium phenotype both in vivo and in vitro: a potential use for bladder defect regeneration
Ming Zhang, Mingxi Xu, Ke Zhang, Zhe Zhou, Yang Zhao, Zhong Wang, Mujun Lu
Abstract
Ming Zhang, Mingxi Xu, Ke Zhang, Zhe Zhou, Yang Zhao, Zhong Wang, Mujun Lu
Abstract
EXPERIMENTAL: ASCs were isolated from the human adipose tissue of individuals undergoing liposuction procedures. Labeling with CM-DiI, the ASCs were mixed with an immortalized urothelium cell line (HUC) and implanted into the subcutaneous tissue of athymic mice for 4 weeks. In vitro, ASCs were induced by conditioned media (CM) and the transwell co-culture with HUC for 21 days. Protein and mRNA expression of the urothelium specific markers UP-1A and UP-II were detected. Array detection of the upper medium was used to screen 41 cytokines and receptors in induced ASCs at different time points. RESULTS: After co-cultured with HUC in vivo for 4 weeks, the percentage of ASCs expressed UP-1A and UP-II increased dramatically compare to co-cultured for 2 weeks. In vitro, after induction for 21 days, ASCs had significantly increased expression of UP-1A and UP-II on protein and mRNA level compare to induction for 7 days. In the CM of urothelium, 28 cytokines and 8 cytokine receptors had significantly higher expression than the non-induced ASCs. After 7 days of induction, the expression of 22 cytokines and 8 cytokine receptors was significantly elevated in induced ASCs compared to non-induced ASCs. At the early and intermediate time points, ASCs secreted high levels of cytokines and soluble receptors, but their expressions decreased significantly at the late time point. CONCLUSION: s: ASCs have the potential to be differentiated into urothelium phenotype both in vivo and in vitro. The cytokines and receptors involved in the differentiation process showed dynamic temporal changes. The results suggest that ASCs may have a potential to be used in urinary tract repair by tissue engineering approach in the future.
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EXPERIMENTAL: ASCs were isolated from the human adipose tissue of individuals undergoing liposuction procedures. Labeling with CM-DiI, the ASCs were mixed with an immortalized urothelium cell line (HUC) and implanted into the subcutaneous tissue of athymic mice for 4 weeks. In vitro, ASCs were induced by conditioned media (CM) and the transwell co-culture with HUC for 21 days. Protein and mRNA expression of the urothelium specific markers UP-1A and UP-II were detected. Array detection of the upper medium was used to screen 41 cytokines and receptors in induced ASCs at different time points. RESULTS: After co-cultured with HUC in vivo for 4 weeks, the percentage of ASCs expressed UP-1A and UP-II increased dramatically compare to co-cultured for 2 weeks. In vitro, after induction for 21 days, ASCs had significantly increased expression of UP-1A and UP-II on protein and mRNA level compare to induction for 7 days. In the CM of urothelium, 28 cytokines and 8 cytokine receptors had significantly higher expression than the non-induced ASCs. After 7 days of induction, the expression of 22 cytokines and 8 cytokine receptors was significantly elevated in induced ASCs compared to non-induced ASCs. At the early and intermediate time points, ASCs secreted high levels of cytokines and soluble receptors, but their expressions decreased significantly at the late time point. CONCLUSION: s: ASCs have the potential to be differentiated into urothelium phenotype both in vivo and in vitro. The cytokines and receptors involved in the differentiation process showed dynamic temporal changes. The results suggest that ASCs may have a potential to be used in urinary tract repair by tissue engineering approach in the future.
Key concepts: Urothelium, Adipose tissue, Regeneration (biology), Liposuction, Urothelial Cell, In vitro, In vivo, Adipocyte