RESUMO
BACKGROUND: Mesenchymal stem cells (MSCs) from human umbilical cord vein have great potential for use in cell therapy because of their ease of isolation, expansion, and differentiation, in addition to their relative acceptance from the ethical point of view. Obtaining the umbilical cord at birth does not present any risk to either mother or child. OBJECTIVE: To isolate and promote in vitro expansion and differentiation of MSCs from human umbilical cord vein into cells with a pancreatic endocrine phenotype. METHODS: Mesenchymal stem cells obtained from human umbilical cord vein via collagenase digestion were characterized at cytochemistry and fluorescent-activated cell sorting, and expanded in vitro. Differentiation of MSCs into an endocrine phenotype was induced using high-glucose (23 mmol/L) medium containing nicotinamide, exendin-4, and 2-mercaptoethanol. Expression of insulin, somatostatin, glucagon, and pancreatic and duodenal homeobox 1 was analyzed using immunofluorescence. RESULTS: Cells isolated from the umbilical cord vein were MSCs as confirmed at cytochemistry and fluorescent-activated cell sorting. Expression of somatostatin, glucagon, and pancreatic and duodenal homeobox 1 by differentiated cells was demonstrated using immunofluorescence. Insulin was not expressed. CONCLUSIONS: The MSC differentiation protocol used in the present study induced expression of some endocrine markers. Insulin was not produced by these cells, probably because of incomplete induction of differentiation.
Assuntos
Ilhotas Pancreáticas/fisiologia , Células-Tronco Mesenquimais/fisiologia , Veias Umbilicais/fisiologia , Antígenos CD/análise , Diferenciação Celular , Divisão Celular , Feminino , Glucagon/genética , Proteínas de Homeodomínio/genética , Humanos , Células-Tronco Mesenquimais/citologia , Fenótipo , Gravidez , Somatostatina/genética , Transativadores/genética , Cordão Umbilical , Veias Umbilicais/citologiaRESUMO
BACKGROUND: Mesenchymal stem cells (MSCs) are an attractive source for generation of cells with beta-cell properties. Previous studies have demonstrated the ability of prolactin to induce an increase in beta-cell mass and maturation, which suggests beneficial effects of its use in MSC differentiation protocols. OBJECTIVE: To evaluate the expression of endocrine differentiation markers in rat MSCs treated in vitro with prolactin. METHODS: Mesenchymal stem cells from bone marrow of Wistar rats were isolated, expanded, and characterized. Differentiation of MSCs was induced in medium containing 23 mmol/L of glucose, and nicotinamide, 2-mercaptoethanol, and exendin-4, in the presence or absence of 500 ng/mL of rat recombinant prolactin. Expression of endocrine markers and prolactin receptor genes was evaluated using real-time polymerase chain reaction, and compared between culture stages and presence vs absence of prolactin in the culture medium. Expression of insulin, somatostatin, glucagon, and pancreatic and duodenal homeobox 1 was also evaluated at immunofluorescence microscopy. RESULTS: Isolated cells were mostly MSCs, as confirmed at fluorescent-activated cell sorting and cytochemistry. Pax6, Ngn-3, Isl1, NeuroD1, Nkx2.2, and Nkx6.1 exhibited varied expression during culture stages. The long form of the prolactin receptor messenger RNA was induced in prolactin-treated cultures (P < .05). The somatostatin gene was induced in early stages of differentiation (P < .05), and its expression was induced by prolactin, as confirmed using immunofluorescence. CONCLUSION: Culture of rat bone marrow MSCs in differentiation medium induces expression of pancreatic endocrine-specific genes, and somatostatin and prolactin receptor expression was also induced by prolactin.
Assuntos
Células da Medula Óssea/fisiologia , Diferenciação Celular/fisiologia , Ilhotas Pancreáticas/fisiologia , Células-Tronco Mesenquimais/fisiologia , Prolactina/farmacologia , Adipócitos/citologia , Adipócitos/fisiologia , Animais , Antígenos CD/biossíntese , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Adesão Celular , Técnicas de Cultura de Células , Condrócitos/citologia , Condrócitos/fisiologia , Glucagon/genética , Glucose/farmacologia , Proteína Homeobox Nkx-2.2 , Insulina/genética , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteócitos/citologia , Osteócitos/fisiologia , Polipeptídeo Pancreático/genética , Reação em Cadeia da Polimerase , RNA/genética , RNA/isolamento & purificação , Ratos , Ratos Wistar , Somatostatina/genéticaRESUMO
This study was developed in order to compare the use of the ABBREVIATED INJURY SCALE (AIS) and the CONDENSED ABBREVIATED INJURY SCALE (CAIS) as basis to calculate INJURY SEVERITY SCORE (ISS) in head injured patients. The results showed that the ISS value was equivalent in the majority of the patients (58.51%) codified by both scales. Also no statistic differences between the scales were perceived when we compared the severity levels as severe, moderate and minor, 61.38% of the lesions scored by AIS/90 were scored by CAIS/85, too.
Assuntos
Escala Resumida de Ferimentos , Traumatismos Craniocerebrais/diagnóstico , Grupos Diagnósticos Relacionados/classificação , Escala de Gravidade do Ferimento , Indexação e Redação de Resumos , Adolescente , Adulto , Criança , Traumatismos Craniocerebrais/classificação , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos TestesRESUMO
This paper reports a case of infection by Histoplasma capsulatum apparently restricted to the peritoneum in a woman submitted to continuous ambulatory peritoneal dialysis. Diagnosis was established by culture of dialysis fluid and peritoneal nodule and by histopathologic examination.
