A simplified method to recover urinary vesicles for clinical applications, and sample banking.
Sci Rep
; 4: 7532, 2014 Dec 23.
Article
em En
| MEDLINE
| ID: mdl-25532487
ABSTRACT
Urinary extracellular vesicles provide a novel source for valuable biomarkers for kidney and urogenital diseases Current isolation protocols include laborious, sequential centrifugation steps which hampers their widespread research and clinical use. Furthermore, large individual urine sample volumes or sizable target cohorts are to be processed (e.g. for biobanking), the storage capacity is an additional problem. Thus, alternative methods are necessary to overcome such limitations. We have developed a practical vesicle isolation technique to yield easily manageable sample volumes in an exceptionally cost efficient way to facilitate their full utilization in less privileged environments and maximize the benefit of biobanking. Urinary vesicles were isolated by hydrostatic dialysis with minimal interference of soluble proteins or vesicle loss. Large volumes of urine were concentrated up to 1/100 of original volume and the dialysis step allowed equalization of urine physico-chemical characteristics. Vesicle fractions were found suitable to any applications, including RNA analysis. In the yield, our hydrostatic filtration dialysis system outperforms the conventional ultracentrifugation-based methods and the labour intensive and potentially hazardous step of ultracentrifugations are eliminated. Likewise, the need for trained laboratory personnel and heavy initial investment is avoided. Thus, our method qualifies as a method for laboratories working with urinary vesicles and biobanking.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Preservação Biológica
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Doenças Urológicas
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Bancos de Espécimes Biológicos
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Micropartículas Derivadas de Células
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Doenças dos Genitais Femininos
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Doenças dos Genitais Masculinos
Tipo de estudo:
Guideline
Limite:
Adult
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Female
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Humans
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Male
Idioma:
En
Revista:
Sci Rep
Ano de publicação:
2014
Tipo de documento:
Article