title: Approach toward an efficient inoculum preparation stage for suspension BHK-21 cell culture.
abstract: Mammalian cells are the most frequently used hosts for biopharmaceutical proteins manufacturing. Inoculum quality is a key element for establishing an efficient bioconversion process. The main objective in inoculation expansion process is to generate large volume of viable cells in the shortest time. The aim of this paper was to optimize the inoculum preparation stage of baby hamster kidney (BHK)-21 cells for suspension cultures in benchtop bioreactors, by means of a combination of static and agitated culture systems. Critical parameters for static (liquid column height: 5, 10, 15 mm) and agitated (working volume: 35, 50, 65 mL, inoculum volume percentage: 10, 30 % and agitation speed: 25, 60 rpm) cultures were study in T-flask and spinner flask, respectively. The optimal liquid column height was 5 mm for static culture. The maximum viable cell concentration in spinner flask cultures was reached with 50 mL working volume and the inoculum volume percentage was not significant in the range under study (10-30 %) at 25 rpm agitation. Agitation speed at 60 rpm did not change the main kinetic parameters with respect to those observed for 25 rpm. These results allowed for a schedule to produce more than 4 × 10(9) BHK-21 cells from 4 × 10(6) cells in 13 day with 1,051 mL culture medium.
authors.affiliation: Laboratório de Células Animais, Departamento de Engenharia Química, Escola Politécnica, Universidade de São Paulo, Av. Prof. Luciano Gualberto, trav. 3, 380, Butantã, São Paulo, SP, 05508-900, Brazil. eutimiocu@yahoo.com.|Departamento de Ciências Biológicas, Universidade Estadual "Júlio de Mesquita Filho", Campus-Assis, Avenida Dom Antonio 2100, Bairro Parque Universitário, 19806-900, Assis, SP, Brazil. eutimiocu@yahoo.com.
authors.forename: Eutimio Gustavo
authors.initials: EG
authors.lastname: Fernández Núñez
authors.affiliation: Laboratório Especial de Pesquisa e Desenvolvimento em Imunológicos Veterinários, Instituto Butantan, Av. Vital Brasil, 1500, Butantã, 05503-900, São Paulo, SP, Brazil.
authors.forename: Jaci
authors.initials: J
authors.lastname: Leme
authors.affiliation: Laboratório de Células Animais, Departamento de Engenharia Química, Escola Politécnica, Universidade de São Paulo, Av. Prof. Luciano Gualberto, trav. 3, 380, Butantã, São Paulo, SP, 05508-900, Brazil.
authors.forename: Letícia
authors.initials: L
authors.lastname: de Almeida Parizotto
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil, 1500, Butantã, São Paulo, SP, 05503-900, Brazil.
authors.forename: Alexandre Gonçalves
authors.initials: AG
authors.lastname: de Rezende
authors.affiliation: Laboratório de Biotecnologia Industrial, Núcleo de Bionanomanufatura, Instituto de Pesquisas Tecnológicas do Estado de São Paulo, Av. Prof. Almeida Prado 532 Cid. Universitária - Butantã, São Paulo, SP, 05508-901, Brazil.
authors.forename: Bruno Labate Vale
authors.initials: BLV
authors.lastname: da Costa
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil, 1500, Butantã, São Paulo, SP, 05503-900, Brazil.
authors.forename: Vera Lucia Lopes
authors.initials: VLL
authors.lastname: Boldorini
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil, 1500, Butantã, São Paulo, SP, 05503-900, Brazil.
authors.forename: Soraia Attie Calil
authors.initials: SAC
authors.lastname: Jorge
authors.affiliation: Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil, 1500, Butantã, São Paulo, SP, 05503-900, Brazil.
authors.forename: Renato Mancini
authors.initials: RM
authors.lastname: Astray
authors.affiliation: Laboratório de Células Animais, Departamento de Engenharia Química, Escola Politécnica, Universidade de São Paulo, Av. Prof. Luciano Gualberto, trav. 3, 380, Butantã, São Paulo, SP, 05508-900, Brazil.|Laboratório de Imunologia Viral, Instituto Butantan, Av. Vital Brasil, 1500, Butantã, São Paulo, SP, 05503-900, Brazil.
authors.forename: Carlos Augusto
authors.initials: CA
authors.lastname: Pereira
authors.affiliation: Laboratório Especial de Pesquisa e Desenvolvimento em Imunológicos Veterinários, Instituto Butantan, Av. Vital Brasil, 1500, Butantã, 05503-900, São Paulo, SP, Brazil.
authors.forename: Celso Pereira
authors.initials: CP
authors.lastname: Caricati
authors.affiliation: Laboratório de Células Animais, Departamento de Engenharia Química, Escola Politécnica, Universidade de São Paulo, Av. Prof. Luciano Gualberto, trav. 3, 380, Butantã, São Paulo, SP, 05508-900, Brazil.
authors.forename: Aldo
authors.initials: A
authors.lastname: Tonso
keywords: BHK-21; Bioreactor; Inoculum preparation; Mammalian cells; Spinner flask; Static cell culture; Suspension cell culture