RESUMEN
The scarcity of human organs available for transplantation is clearly evident. Efforts to maximize the use of available organs and to increase the number of donors have increased the number of transplantations performed, but at a rate that remains far behind the rate of growth of the waiting list. Thus, the likelihood of a patient with severe liver disease receiving a liver replacement is decreasing. In order to offer treatment to most patients with liver disease, alternatives to whole-organ replacement must be found. Cell-based treatments, in which suspensions of liver cells are injected into patients with liver failure and reconstitute the patient's liver functions, may be that alternative. Here, we report on a regulatory-compliant process for the production of a cryopreserved cell therapy product that yields viable, metabolically active hepatocytes that can be infused directly into patients with the goal of reconstituting liver function.
Asunto(s)
Hepatocitos/trasplante , Fallo Hepático/terapia , Separación Celular/métodos , Supervivencia Celular , Trasplante de Células/métodos , Ensayos Clínicos Fase I como Asunto , Cumarinas/metabolismo , Criopreservación/métodos , Epítopos , Citometría de Flujo/métodos , Hepatocitos/metabolismo , Humanos , Selección de Paciente , Recolección de Tejidos y Órganos/métodos , Obtención de Tejidos y Órganos , Trasplante Homólogo/métodos , Estados Unidos , United States Food and Drug Administration , Urea/metabolismoRESUMEN
Activity of cytochrome p450 (CYP450) enzyme is used to measure the ability of hepatocytes to metabolize pharmaceutical compounds. When determining functionality of hepatocytes, the cells may be induced in order to determine metabolic activity after drug induction. Hepatocytes, whether in suspension or plated, are used in the pharmaceutical industry as a surrogate to assess in vivo drug metabolism. Within this chapter, isolation of hepatocytes from whole-liver tissue and subsequent preparation for CYP450 istotype 3A4 activity is discussed.