RESUMO
Enhancing the proliferative capacity of mesenchymal stem cells (MSCs) is critical for increasing their therapeutic potential in a variety of diseases. We hypothesized that lentivirus-mediated overexpression of canine octamer-binding transcription factor 4 (OCT4) might influence the proliferation of canine adipose tissue-derived MSCs (cATMSCs). cOCT4-cATMSCs were generated by transducing cATMSCs with a cOCT4-lentiviral vector. Increased expression of cOCT4 was confirmed using RT-PCR and immunoblotting. Immunophenotypic characterization using flow cytometry indicated that the CD29, CD44, CD73, CD90, and CD105 surface markers were highly expressed by both cOCT4- and mock-transduced cATMSCs (mock-cATMSCs), whereas the CD31 and CD45 markers were absent. We performed the osteogenic differentiation assay to evaluate the effects of cOCT4 overexpression on the osteogenic differentiation potential of cATMSCs. The results showed that cOCT4-cATMSCs had a much higher potential for osteogenic differentiation than mock-cATMSCs. Next, the proliferative capacities of cOCT4- and mock-cATMSCs were evaluated using a WST-1 cell proliferation assay and trypan blue exclusion. cOCT4-cATMSCs showed a higher proliferative capacity than mock-cATMSCs. Cell cycle analysis indicated that overexpression of cOCT4 in cATMSCs induced an increase in the proportion of cells in S and G2/M phases. Consistent with this, immunoblot analysis showed that cyclin D1 expression was increased in cOCT4-cATMSCs. In conclusion, our results indicate that lentivirus-mediated overexpression of cOCT4 increased the proliferative capacity of cATMSCs. OCT4-mediated enhancement of cell proliferation may be a useful method for expanding MSC population rapidly without loss of stemness.
Assuntos
Tecido Adiposo/citologia , Células-Tronco Mesenquimais/citologia , Fator 3 de Transcrição de Octâmero/metabolismo , Tecido Adiposo/metabolismo , Animais , Antígenos CD/metabolismo , Diferenciação Celular , Proliferação de Células/genética , Células Cultivadas , Ciclina D1/metabolismo , Cães , Fase G2 , Vetores Genéticos/metabolismo , Lentivirus/genética , Células-Tronco Mesenquimais/metabolismo , Fator 3 de Transcrição de Octâmero/genética , Osteogênese , Fase SRESUMO
With the advent of small-molecule immune modulators, recombinant fusion proteins, and monoclonal antibodies, treatment options for patients with rheumatic diseases are now broad. These agents carry significant risks and an individualized approach to each patient, balancing known risks and benefits, remains the most prudent course. This review summarizes the available immunosuppressant treatments, discusses their perioperative implications, and provides recommendations for their perioperative management.
Assuntos
Imunossupressores , Doenças Reumáticas , Humanos , Doenças Reumáticas/tratamento farmacológico , Imunossupressores/uso terapêutico , Cuidados Pré-Operatórios/métodosRESUMO
With the advent of small-molecule immune modulators, recombinant fusion proteins, and monoclonal antibodies, treatment options for patients with rheumatic diseases are now broad. These agents carry significant risks and an individualized approach to each patient, balancing known risks and benefits, remains the most prudent course. This review summarizes the available immunosuppressant treatments, discusses their perioperative implications, and provides recommendations for their perioperative management.
Assuntos
Artrite Reumatoide , Doenças Reumáticas , Humanos , Imunossupressores/uso terapêutico , Doenças Reumáticas/tratamento farmacológico , Doenças Reumáticas/cirurgiaRESUMO
BACKGROUND AIMS: Adipose tissue (AT)-derived mesenchymal stromal cells (MSC) (AT-MSC) represent a novel tool for delivering therapeutic genes to tumor cells. Interferon (IFN)-ß is a cytokine with pleiotropic cellular functions, including anti-proliferative, immunomodulatory and anti-angiogenic activities. The purpose of this study was to engineer canine AT-MSC (cAT-MSC) producing IFN-ß and to evaluate the anti-tumor effect of cAT-MSC-IFN-ß combined with cisplatin in mouse melanoma model. METHODS: cAT-MSC engineered to express mouse IFN-ß were generated using a lentiviral vector (cAT-MSC-IFN-ß) and the secreted IFN-ß-induced inhibition of tumor cell growth and apoptosis on B16F10 cells was investigated in vitro prior to in vivo studies. Melanoma-bearing mouse was developed by injecting B16F10 cells subcutaneously into 6-week-old C57BL/6 mice. After 14 days, cisplatin (10 mg/kg) was injected intratumorally, and 3 days later the engineered cAT-MSC were injected subcutaneously every 3 days to death. Tumor volume and survival times were measured. RESULTS: The combination treatment of cAT-MSC-IFN-ß with cisplatin was more effective in inhibiting the growth of melanoma and resulted in significantly extended survival time than both an unengineered cAT-MSC-cisplatin combination group and a cisplatin-alone group. Interestingly, subcutaneously injected cAT-MSC-IFN-ß were migrated to tumor sites. CONCLUSIONS: Our data suggest that canine AT-MSC could serve as a powerful cell-based delivery vehicle for releasing therapeutic proteins to tumor lesions. Maximal anti-tumor effects were seen when this therapy was combined with a DNA-damaging chemotherapeutic agent. This study demonstrates the possible applicability of AT-MSC-mediated IFN-ß in treating canine and human cancer patients.