RESUMEN
BACKGROUND: High-quality biospecimens of human origin with annotated clinical and procedural data are an important tool for biomedical research, not only to map physiology, pathophysiology and aetiology but also to go beyond in translational research. This has opened a new special field of research known as 'biobanking', which focuses on how to collect, store and provide these specimens and data, and which is substantially supported by national and European funding. PURPOSE: An overview on biobanking is given, with a closer look on a clinical setting, concerning a necessary distinction from clinical trials and studies as well as a comparison of prospective sample collection with secondary use of archived samples from diagnostics. Based on a summary of possible use and scientific impact of human tissue in research, it is shown how surgical expertise boosts the scientific value of specimens and data. Finally, an assessment of legal and ethical issues especially from a surgical perspective is given, followed by a model of interdisciplinary biobanking within a joint 'centre' that as synergistic structure merges essential input from surgery as well as laboratory medicine, pathology and biometry. CONCLUSION: Within the domain of biobanking, surgeons have to develop a better awareness of their role within translational research, not only on the level of medical faculties but also as nationally and internationally funded initiatives. Therefore, the authors suggest a platform for biobanking within the German association of surgeons in analogy to the existing special interest group for clinical trials.
Asunto(s)
Bancos de Muestras Biológicas/organización & administración , Conducta Cooperativa , Recolección de Datos , Cirugía General/organización & administración , Comunicación Interdisciplinaria , Rol del Médico , Investigación Biomédica Traslacional/organización & administración , Bancos de Muestras Biológicas/legislación & jurisprudencia , Recolección de Datos/legislación & jurisprudencia , Europa (Continente) , Cirugía General/legislación & jurisprudencia , Humanos , Consentimiento Informado/legislación & jurisprudencia , Apoyo a la Investigación como Asunto/legislación & jurisprudencia , Apoyo a la Investigación como Asunto/organización & administración , Recolección de Tejidos y Órganos/legislación & jurisprudencia , Investigación Biomédica Traslacional/legislación & jurisprudenciaAsunto(s)
Bancos de Muestras Biológicas/tendencias , Cirugía General/tendencias , Bancos de Muestras Biológicas/ética , Bancos de Muestras Biológicas/organización & administración , Investigación Biomédica/ética , Investigación Biomédica/organización & administración , Investigación Biomédica/tendencias , Predicción , Cirugía General/ética , Alemania , Humanos , PolíticaRESUMEN
Co-transplantation of hepatocytes and hepatic stellate cells has been shown to increase the engraftment of transplanted hepatocytes in the liver. Here, we describe a method for the simultaneous isolation of human primary hepatocytes and hepatic stellate cells from the same donor for co-transplantation or for use in in vitro cell culture models.
Asunto(s)
Separación Celular/métodos , Células Estrelladas Hepáticas/fisiología , Hepatocitos/fisiología , Hígado/cirugía , Perfusión/métodos , Separación Celular/instrumentación , Supervivencia Celular , Trasplante de Células/métodos , Células Cultivadas , Técnicas de Cocultivo/métodos , Criopreservación , Células Estrelladas Hepáticas/trasplante , Hepatocitos/trasplante , Humanos , Hígado/citología , Perfusión/instrumentación , Cultivo Primario de Células/métodos , Donantes de TejidosRESUMEN
Isolated human primary hepatocytes are an essential in vitro model for basic and clinical research. For successful application as a model, isolated hepatocytes need to have a good viability and be available in sufficient yield. Therefore, this study aims to identify donor characteristics, intra-operative factors, tissue processing and cell isolation parameters that affect the viability and yield of human hepatocytes. Remnant liver pieces from tissue designated as surgical waste were collected from 1034 donors with informed consent. Human hepatocytes were isolated by a two-step collagenase perfusion technique with modifications and hepatocyte yield and viability were subsequently determined. The accompanying patient data was collected and entered into a database. Univariate analyses found that the viability and the yield of hepatocytes were affected by many of the variables examined. Multivariate analyses were then carried out to confirm the factors that have a significant relationship with the viability and the yield. It was found that the viability of hepatocytes was significantly decreased by the presence of fibrosis, liver fat and with increasing gamma-glutamyltranspeptidase activity and bilirubin content. Yield was significantly decreased by the presence of liver fat, septal fibrosis, with increasing aspartate aminotransferase activity, cold ischemia times and weight of perfused liver. However, yield was significantly increased by chemotherapy treatment. In conclusion, this study determined the variables that have a significant effect on the viability and the yield of isolated human hepatocytes. These variables have been used to generate an algorithm that can calculate projected viability and yield of isolated human hepatocytes. In this way, projected viability can be determined even before isolation of hepatocytes, so that donors that result in high viability and yield can be identified. Further, if the viability and yield of the isolated hepatocytes is lower than expected, this will highlight a methodological problem that can be addressed.
Asunto(s)
Algoritmos , Separación Celular/métodos , Hígado/citología , Factores de Edad , Aspartato Aminotransferasas/metabolismo , Bilirrubina/metabolismo , Índice de Masa Corporal , Supervivencia Celular , Células Cultivadas , Colagenasas/metabolismo , Femenino , Fibrosis/patología , Humanos , Hígado/metabolismo , Masculino , Factores Sexuales , Donantes de Tejidos , gamma-Glutamiltransferasa/metabolismoRESUMEN
The accuracy of information garnered by real-time quantitative polymerase chain reaction (RT-qPCR), an important technology for elucidating molecular mechanisms of disease, is dependent on tissue quality. Thus, this study aimed to determine the effects of intra-operative manipulation, extended processing times, different temperatures or storage in RNAlater on RNA quality in liver samples for tissue banking. Liver samples, flash-frozen or in RNAlater, were collected over a time course (during surgery before blood arrest up to 1 day after surgery) with samples kept either at room temperature (RT) or on ice. This study showed that at the longest time-point at RT, the RNA quality decreased significantly by 20%. However, relative gene expressions of FOS, GUSB, MYC, HIF1α and GFER were in general not significantly different when the time-points were compared. In conclusion, samples should be kept on ice during processing, and either RNAlater or snap-freezing should be utilised for storage. Further, intra-operative manipulation and extended postoperative processing time generally does not change relative gene expression levels for the 5 genes studied, making such sampling suitable for RT-qPCR analysis. Thus, if relative gene expression of a gene of interest is stable, these guidelines will lead to increased accrual of samples to the tissue bank.