ARTICULOS ORIGINALES

 

 

Ensayo in vivo para determinar agentes extraños en células BHK-21 empleadas en la obtención de biológicos.

 

In vivo determination of adventitious agents in BHK-21 cells used in the production of
biologicals.

 

Georgina Pardo, Ernesto Almora, Odalys Fidalgo, Arelys Zamora, Niurka Rodríguez y Ela María Pérez.

Instituto Finlay, Centro de Investigación-Producción de Vacunas y Sueros. Ciudad de La Habana, Cuba. E-mail:
[e-mail]

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RESUMEN

La controversia sobre los parámetros de calidad de las células de línea como substrato para la obtención de
productos farmacéuticos ha sido un tema de interés desde 1987. Las pruebas realizadas a las células, generalmente siguen las recomendaciones de los “Puntos a considerar en la Caracterización de Líneas Celulares empleadas para la Producción de Biológicos” entre las que se encuentra que estén libres de agentes adventicios. En este estudio al clon 13 de células BHK-21 procedente de riñón de hámster sirio recién nacido, se le determina la presencia de agentes extraños in vivo; así como su inocuidad en animales de experimentación. La prueba in vivo incluyó la inoculación de embriones de pollo por cavidad alantoidea y saco vitelino, ratones lactantes y adultos, curieles y conejos. Todos los animales fueron observados para determinar signos clínicos de infección viral. En los embriones se observó la viabilidad. En todos los casos se mantuvo el 100% de los embriones y animales vivos y saludables durante el período de observación. Las células del banco estaban libres de virus hemaglutinantes y hemadsorbentes, virus de la coriomeningitis linfocítica y coxsackie. Los pesos de los animales inoculados fueron mayores que los de los animales controles. Las células no presentaron evidencia de agentes extraños en los ensayos in vivo y fueron bien toleradas e inocuas en los animales de experimentación.

Palabras claves: Caracterización, células BHK-21, agentes extraños, animales de experimentación

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ABSTRACT

Acceptance of cell lines as a substrate for production of pharmaceuticals has been a controversial topic since 1987. The cells are generally tested according to the recommendations that appear in “Points to Consider in the
Characterization of Cell Lines Used to Produce Biologicals”. They should be free from adventitious agents; among others in vitro and in vivo tests are recommended for this purpose. In this report BHK-21-cell clone 13 from new born Syrian hamster kidney cells is assayed in vivo to determine adventitious agents; it is also tested for nonspecific innocuousness in experimental animals. In vivo tests included the inoculation of adult mice,suckling mice, guinea pigs, rabbits, and embryonated chicken eggs by the yolk sac and allantoic routes of inoculation. All animals were observed for clinical signs of viral infection. Egg viability was determined. The results showed that 100% of the animals and eggs remained healthy and survived the observation period. The cell bank was free of haemadsorbent, haemagglutinating, lymphocytic chroriomeningitis and Coxsakie viruses. The weights of the tested animals were higher than the weights of the control animals. The cells did not present evidences of adventitious agents in the in vivo assays and they were well accepted by the experimental animals.

Key words: Characterization, BHK-21 cells, adventitious agents, experimental animals.

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