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1.
Tumour Biol ; 36(7): 5667-77, 2015 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-25697898

RESUMEN

Tumor microenvironment is an important factor, which sustains and promotes the tumors by inflammatory signals. Interleukin-6 (IL-6) is known as a multifunctional cytokine, which is a major activator of the signaling pathway of Janus kinases (JAKs)/signal transducer and activator of transcription 3 (STAT3). In this study, we aimed to investigate the effect of IL-6 in the tumor microenvironment on carcinogenesis. For this purpose, healthy breast tissue-derived stromal cells (HBT-SCs) and malign breast tissue-derived stromal cells (MBT-SCs) were co-cultured with MCF-7 (human breast adenocarcinoma cell line) cells using semipermeable membranes. The cell proliferation was monitored with water-soluble tetrazolium (WST) and carboxyfluorescein succinimidyl ester (CFSE) assays. Protein levels were measured by enzyme-linked immunosorbent assay (ELISA) and Western blot hybridization, while gene expressions were measured by real-time PCR. The results demonstrated that IL-6 protein levels increased significantly in the supernatants of MBT-SCs when they were co-cultured with MCF-7 cells. In accordance with this, the expression of IL-6 was significantly higher in MBT-SCs. Additionally, the expression of STAT3 in MCF-7 cells increased slightly when they were co-cultured with MBT-SCs. Considering together, there is an important interaction between tumor microenvironment and tumor cells mediated by IL-6 signaling. Thereby, the targeting on IL-6 signaling in the treatment of cancer might effectively prevent the tumor progression.


Asunto(s)
Neoplasias de la Mama/genética , Interleucina-6/biosíntesis , Factor de Transcripción STAT3/biosíntesis , Microambiente Tumoral/genética , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Carcinogénesis/genética , Carcinogénesis/metabolismo , Proliferación Celular/genética , Técnicas de Cocultivo , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Interleucina-6/genética , Células MCF-7 , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/patología , Factor de Transcripción STAT3/genética , Transducción de Señal
2.
J Periodontol ; 86(2): 283-91, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25325708

RESUMEN

BACKGROUND: Cell-based therapy using mesenchymal stem cells (MSCs) seems promising to obtain regeneration of dental tissues. A comparison of tissue sources, including periodontal ligament (PDL) versus pulp (P), could provide critical information to select an appropriate MSC population for designing predictable regenerative therapies. The purpose of this study is to compare the proliferation and stemness and the MSC-specific and mineralized tissue-specific gene expression of P-MSCs and PDL-MSCs. METHODS: MSCs were obtained from PDL and P tissue of premolars (n = 3) extracted for orthodontic reasons. MSC proliferation was evaluated using a real-time cell analyzer for 160 hours. Telomerase activity was evaluated by a telomeric repeat amplification protocol assay based on enzyme-linked immunosorbent assay. Total RNA was isolated from the MSCs on day 3. A polymerase chain reaction (PCR) array was used to compare the expression of MSC-specific genes. The expression of mineralized tissue-associated genes, including Type I collagen (COL I), runt-related transcription factor 2 (RunX2), bone sialoprotein (BSP), and osteocalcin (OCN) messenger RNA (mRNA), was evaluated using quantitative real-time PCR. RESULTS: Higher proliferation potential and telomerase activity were observed in the P-MSCs compared to PDL-MSCs of premolar teeth. Fourteen of 84 genes related to MSCs were expressed differently in the PDL-MSCs versus the P-MSCs. The expressions of bone morphogenetic protein 2 (BMP2) and BMP6; sex-determining region Y-box 9 (SOX9); integrin, alpha 6 (ITGA6); melanoma cell adhesion molecule (MCAM); phosphatidylinositol glycan anchor biosynthesis, class S (PIGS); prominin 1 (PROM1); ribosomal protein L13A (RPL13A); and microphthalmia-associated transcription factor (MITF) were higher in the P-MSCs compared to the PDL-MSCs, and higher expression of matrix metalloproteinase 2 (MMP2), interleukin (IL)-6, insulin (INS), alanyl (membrane) aminopeptidase (ANPEP), and IL-10 were observed in the PDL-MSCs. However, there was no statistically significant difference in the expression of mineralized tissue-associated genes, including BSP and RunX2, between the P-MSCs and the PDL-MSCs. Higher expression of COL I and lower expression of OCN mRNA transcripts were noted in the PDL-MSCs compared to the P-MSCs. CONCLUSIONS: The results of this study suggest that MSCs isolated from P and PDL tissues show different cellular behavior. To increase the predictability of MSC-based regenerative treatment, differences in dental tissue-derived MSCs and favorable aspects of cell sources should be further clarified.


Asunto(s)
Pulpa Dental/citología , Células Madre Mesenquimatosas/fisiología , Ligamento Periodontal/citología , Antígeno AC133 , Aciltransferasas/análisis , Adolescente , Adulto , Antígenos CD/análisis , Proteína Morfogenética Ósea 2/análisis , Antígenos CD13/análisis , Antígeno CD146/análisis , Proliferación Celular , Separación Celular , Colágeno Tipo I/análisis , Subunidad alfa 1 del Factor de Unión al Sitio Principal/análisis , Femenino , Glicoproteínas/análisis , Humanos , Insulina/análisis , Integrina alfa6/análisis , Sialoproteína de Unión a Integrina/análisis , Interleucina-10/análisis , Interleucina-6/análisis , Metaloproteinasa 2 de la Matriz/análisis , Células Madre Mesenquimatosas/enzimología , Factor de Transcripción Asociado a Microftalmía/análisis , Osteocalcina/análisis , Péptidos/análisis , Proteínas Ribosómicas/análisis , Factor de Transcripción SOX9/análisis , Telomerasa/análisis , Adulto Joven
3.
J Orthop Res ; 32(1): 151-8, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-24115219

RESUMEN

In this study, analysis and comprehensive comparison of neurogenic differentiation capacity of human bursal tissue-derived-stem cells (hBT-SCs) was aimed with human bone marrow derived mesenchymal stem cells (hBM-MSCs). hBT-SCs was isolated from subacromial bursa tissue (n = 3) by collagen type-II digestion. The expression of stem cell markers, differentiation capacity and telomerase activity were determined for both cell lines. The expression levels of neurogenic cell markers were compared consecutively. With respect to the surface marker profile, both cells display similar pluripotency phenotypes. Both cells successfully differentiated into osteo- and adipogenic cell lines. The immune staining of mesenchymal, stem cell and neurogenic markers gave positive reaction. The gene expression level for Tubb3, Nestin, Gfap, Map2, Nf-h, and Nf-l was higher in hBT-SCs than hBM-MSCs. The high level of neurotrophic factors, like Tenascin C, NGF, BDNF, VEGF, and CNTF might indicate their regeneration and maintenance capacity in damaged neural tissue. Besides they are alternative source for human mesenchymal stem cells, hBT-SCs assess the possibility to use in clinical studies.


Asunto(s)
Articulación Acromioclavicular/citología , Bolsa Sinovial/citología , Células Madre Mesenquimatosas/citología , Neurogénesis/fisiología , Neuronas/citología , Biomarcadores/metabolismo , Células de la Médula Ósea/citología , Diferenciación Celular/fisiología , Células Cultivadas , Expresión Génica/fisiología , Humanos , Células Madre Mesenquimatosas/fisiología , Factores de Crecimiento Nervioso/genética , Factores de Crecimiento Nervioso/metabolismo , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Neuroglía/citología , Neuronas/fisiología
4.
Cytotherapy ; 15(5): 557-70, 2013 May.
Artículo en Inglés | MEDLINE | ID: mdl-23388582

RESUMEN

BACKGROUND AIMS: Differentiation or reprogramming of stem cells could be achieved by remodulating the microenvironment, which regulates the fate of cells by soluble factors and contacts. By providing an in vivo-like microenvironment, directional and functional differentiation of stem cells could be achieved in vitro. In this study, the differentiation of mesenchymal stromal cells (MSCs) derived from rat tissues (adipose, rAT; bone marrow, rBM) were analyzed by in vitro and in vivo co-culture experiments. The insulin-producing capacities of islets transplanted under the renal kidney capsule with rAT- and rBM-MSCs were compared and the reduction of hyperglycemia symptoms in rat models was examined. METHODS: MSCs prelabeled with green fluorescence protein were co-cultured with islets directly. The insulin production of cells was determined by immunostaining and ELISA. Streptozotocin-induced diabetic rat models were created and MSCs were co-transplanted with the islets under the kidney capsule to confirm the in vitro results. RESULTS: MSCs were differentiated into insulin-producing cells after 38 days of co-culture, confirmed by insulin and C-peptide stainings. In vivo functional studies revealed that the co-culture of islets with MSCs provided higher differentiation efficiency. The weight gain measurement and glucose tolerance test in the rat group co-transplanted of rAT-MSCs and islets indicate a better recovery than islet-alone transplants and co-transplants of islets and rBM-MSCs. CONCLUSIONS: rAT-MSCs could be considered as the cell of choice for cell-based treatment of type 1 diabetes. Because the co-transplantation of islets with MSCs increases the number of insulin-producing cells, this method was suggested for clinical applications.


Asunto(s)
Tejido Adiposo/citología , Diabetes Mellitus Tipo 1/terapia , Trasplante de Islotes Pancreáticos , Islotes Pancreáticos/citología , Células Madre Mesenquimatosas/citología , Animales , Glucemia/metabolismo , Diferenciación Celular , Células Cultivadas , Técnicas de Cocultivo , Diabetes Mellitus Experimental/terapia , Proteínas Fluorescentes Verdes , Insulina/metabolismo , Secreción de Insulina , Células Secretoras de Insulina/citología , Células Secretoras de Insulina/metabolismo , Islotes Pancreáticos/metabolismo , Trasplante de Células Madre Mesenquimatosas/métodos , Ratas , Nicho de Células Madre
5.
Cytotherapy ; 13(10): 1205-20, 2011 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-21905956

RESUMEN

BACKGROUND AIMS. Studies performed using human and animal models have indicated the immunoregulatory capability of mesenchymal stromal cells in several lineages. We investigated whether human dental pulp-derived stem cells (hDP-SC) have regulatory effects on phytohemagglutinin (PHA)-activated CD3(+) T cells. We aimed to define the regulatory mechanisms associated with hDP-SC that occur in mixed lymphocyte reaction (MLR) and transwell systems with PHA-CD3(+) T cells and hDP-SC at a ratio of 1:1. METHODS. Proliferation, apoptosis and pro- and anti-inflammatory cytokines of PHA-CD3(+)T cells, the expression of Regulatory T cells (Treg) markers and some regulatory factors related to hDP-SC, were studied in Both transwell and MLR are co-cultures systems. RESULTS. Anti-proliferative and apoptotic effects of hDP-SC were determined in co-culture systems. Elevated expression levels of human leukocyte antigen (HLA)-G, hepatocyte growth factor (HGF)-ß1, intracellular adhesion molecule (ICAM-1)-1, interleukin (IL)-6, IL-10, transforming growth factor (TGF)-ß1, vascular adhesion molecule (VCAM)-1 and vascular endothelial growth factor (VEGF) by hDP-SC were detected in the co-culture systems. We observed decreased expression levels of pro-inflammatory cytokines [interferon (IFN)-γ, IL-2, IL-6 receptor (R), IL-12, Interleukin-17A (IL-17A), tumor necrosis factor (TNF)-α] and increased expression levels of anti-inflammatory cytokine [inducible protein (IP)-10] from PHA-CD3(+) T cells in the transwell system. Expression of Treg (CD4(+) CD25(+) Foxp3(+)) markers was significantly induced by hDP-SC in both co-culture systems. We observed apoptosis of PHA-CD3(+) T cells with 24 h using time-lapse camera photographs and active caspase labeling; it is likely that paracrine soluble factors and molecular signals secreted by hDP-SC led this apoptosis. CONCLUSIONS. We suggest that hDP-SC have potent immunoregulatory functions because of their soluble factors and cytokines via paracrine mechanisms associated with PHA-CD3(+) T cells, which could contribute to clinical therapies.


Asunto(s)
Células Madre Adultas/metabolismo , Pulpa Dental/citología , Células Madre Mesenquimatosas/metabolismo , Subgrupos de Linfocitos T/metabolismo , Linfocitos T Reguladores/metabolismo , Adulto , Células Madre Adultas/citología , Células Madre Adultas/inmunología , Antígenos de Diferenciación/metabolismo , Apoptosis , Proliferación Celular , Células Cultivadas , Técnicas de Cocultivo , Citocinas/metabolismo , Humanos , Inmunomodulación , Prueba de Cultivo Mixto de Linfocitos , Masculino , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/inmunología , Comunicación Paracrina , Nicho de Células Madre , Subgrupos de Linfocitos T/citología , Subgrupos de Linfocitos T/inmunología , Linfocitos T Reguladores/citología , Linfocitos T Reguladores/inmunología
6.
Int J Pediatr Otorhinolaryngol ; 68(11): 1399-406, 2004 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-15488971

RESUMEN

OBJECTIVE: The aim of this study was to determine the syndromic etiology of bilateral severe sensorineural hearing disorders in children and current etiological causes to reduce the cases in the unknown group. METHODS: This study was conducted on 550 students of five schools for the deaf in Istanbul and Zonguldak, Turkey. Otologic, audiologic, dysmorphologic, ophthalmologic and dental examinations were performed in all children. Familial and medical histories were obtained. RESULTS: The etiology of hearing loss was genetic in 346 (62.90%), acquired in 107 (19.45%) and unknown in 97 (%17.63) cases. A total of 619 malformations were defined in 550 children and 99 of them belonged to a syndrome. We identified 33 different syndromes for these 99 syndromic children. Syndromic etiology was found in 18.0% of the total and 28.61% of the subjects with genetic etiology. Most common syndrome was Waardenburg syndrome which occurred in 33 children. CONCLUSION: The incidence of hereditary hearing impairment is very high in developing countries compared to developed countries. Prevention is essential to reduce the incidence, multidisciplinary approach and genetic counselling are necessary in this regard.


Asunto(s)
Pérdida Auditiva Bilateral/etiología , Pérdida Auditiva Sensorineural/etiología , Anomalías Múltiples/epidemiología , Adolescente , Adulto , Audiometría , Niño , Consanguinidad , Niños con Discapacidad/educación , Educación de Personas con Discapacidad Auditiva , Femenino , Pérdida Auditiva Bilateral/epidemiología , Pérdida Auditiva Sensorineural/epidemiología , Humanos , Masculino , Instituciones Académicas , Síndrome , Turquía/epidemiología
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