Pre-Clinical Efficacy and Safety Evaluation of Human Amniotic Fluid-Derived Stem Cell Injection in a Mouse Model of Urinary Incontinence

PURPOSE: Stem cell-based therapies represent new promises for the treatment of urinary incontinence. This study was performed to assess optimized cell passage number, cell dose, therapeutic efficacy, feasibility, toxicity, and cell trafficking for the first step of the pre-clinical evaluation of human amniotic fluid stem cell (hAFSC) therapy in a urinary incontinence animal model. MATERIALS AND METHODS: The proper cell passage number was analyzed with hAFSCs at passages 4, 6, and 8 at week 2. The cell dose optimization included 1x10(4), 1x10(5), and 1x10(6) cells at week 2. The in vivo cell toxicity was performed with 0.25x10(6), 0.5x10(6), and 1x10(6) cells at weeks 2 and 4. Cell tracking was performed with 1x10(6) cells at weeks 2 and 4. RESULTS: The selected optimal cell passage number was smaller than 6, and the optimal cell dose was 1x10(6) for the mouse model. In our pre-clinical study, hAFSC-injected animals showed normal values for several parameters. Moreover, the injected cells were found to be non-toxic and non-tumorigenic. Furthermore, the injected hAFSCs were rarely identified by in vivo cell trafficking in the target organs at week 2. CONCLUSION: This study demonstrates for the first time the pre-clinical efficacy and safety of hAFSC injection in the urinary incontinence animal model and provides a basis for future clinical applications.

Cryopreservation of Collected Peripheral Blood Hematopoietic Stem Cell Product with 5% DMSO by Adding Nontoxic Natural Cryoprotectants / 대한수혈학회지

BACKGROUND: Cryopreservation of hematopoietic stem cells has become an important process due to the therapeutic protocol, which includes stem cell transplantation after chemotherapy, for many hematological malignancies. The conventional medium contains 10% dimethyl sulfoxide (DMSO) as a cryoprotectant, but this has been reported to be related with many complications. We analyzed the usefulness of trehalose, catalase and zVAD-fmk for cryopreservation along with using a reduced concentration of DMSO to 5%. METHODS: Peripheral blood stem cells were frozen in 10% DMSO as a control and also in 5% DMSO with trehalose and catalase. After 3 weeks of storage in a liquid nitrogen tank, the viability of the thawed hematopoietic stem cells was measured using Trypan blue staining and 7-AAD analysis via conducting flow cytometry. The colony forming potential was assessed using methylcellulose culture. We measured the viability of cells in 5% DMSO medium with or without addition of 30 uM zVAD-fmk right after thawing, and we also did this 6 and 24 hours after incubation. RESULTS: Cryopreserved cells in 5% DMSO with trehalose and catalase showed similar survival (50.42%) compared with the control (49.78%). The viability of cells that were also treated with added zVAD-fmk showed a better result (13.12%) than without it (5.5%) after 24 hours of incubation. Colony forming assay showed similar colony formation in 5% DMSO with the natural cryoprotectants. CONCLUSION: According to the results, lowering the DMSO concentration to 5% is significant and we can expect better cell viability and prevent many side effects of high dose DMSO when adding natural cryprotectants in the cryopreservation medium or by adding caspase-inhibitor right after thawing.

Effect of Chitosan Oligosaccharide on Enzymes for Cancer Chemoprevention / 대한암학회지

PURPOSE: Two types of chitosan oligosaccharides (COSs), COS I and COS II, were investigated for the effects on ascitic tumor and enzymes for cancer chemoprevention. MATERIALS AND METHODS: Chitosan oligosaccharides were administered once daily for 10 days after the tumor implantation. The change of body weight was observed for 20 days, and the survival rate of mice was determined after 21 days. Chitosan oligosaccharides were administered once daily for 10 days before the tumor implantation (1 106 cells). The number of ascitic tumor cells were measured at 6 days after tumor implantation. Chemopreventive potential of chitosan oligosaccharides was examined by the induction of quinone reductase and inhibition of cytochrome P450 1A1. RESULTS: Chitosan oligosaccharides exerted antitumor activity by inhibiting the growth of Ehrlich ascites tumor cells in vivo. Mice given Ehrlich cells and 10 or 100 mg/kg body weight of chitosan oligosaccharides had 33% survival after 21 days. Quinone reductase activity was increased with chitosan oligosaccharides. There were 26% and 33% inhibition in the activity of cytochrome P450 1A1 enzyme with the treatment of COS I and COS II, respectively. CONCLUSION: These results suggest that chitosan oligosaccharides has antitumor activity and cancer chemo preventive potential by inducing QR activity and inhibiting cytochrome P450 1A1.

Genetic Safety Study of Chlorpromazine / 신경정신의학

OBJECT: The aim of this study is to determine whether exposure to chlorpromazine causes mutagenicity and genetic disorders. METHOD: Ames (Salmonella typhimurium) test and Rec assay (Bacillus subtilis) were used as indicators for DNA damage. Furthermore, the levels of umu operon expression by measuring the beta-galactosidase activity were monitered with the SOS umu test using S. typhimurium 1535 containing plasmid pSK1002. And the host-mediated assay was used to investigate the muta-genicity of chlorpromazine after the activation with in vivo metabolic systems. RESULTS: From the results, chlorpromazine did not affect DNA of S. typhimurium and B. subtilis strains and showed no mutagenicity at the all concentrations tested. These phenomena was also similar to that after metabolic activation of chlorpromazine in in vivo system. CONCLUSION: These results suggested that chlorpromazine did not show the mutagenicity and genotoxicity by four different methods used in this study.

Effect of Interleukin-2 on the Surgically Induced Enndometriosis in Rat / 대한면역학회지

It has been shown that wornen with endometriosis have several immunological defects. The effect of interleukin-2 (IL-2) for the treatment of induced endometriosis in rat was studied. The results obtained are as followings: proliferation of epithelium is increased, and the inner surface is undulated with 1.5 nM IL-2. In 7.5 nM IL-2, the epithelial cells are changed to columar ones, and secretory hobs are observed at the apex of individual cell. Secretory activity of epithelium is increased with 0.5 nM IL-2, and apoptosis of the epithelial cell is observed in 15 nM IL-2. The levels of progesterone and estradiol in sera of rat were increased after treatment with IL-2 and were highest in the concentration of 1.5 nM IL-2. The results of this study can be a guide in the development of new therapeutic approaches for the treatment of endometriosis.