Abstract

The fact of microcarrier (MC) culture introduces new possibilities and makes possible the practical high-yield culture of anchorage-dependent cells has generated a considerable focus in this study. The objective of this research was to study the comparison of Vero cell growth on different types of commercial microcarriers; Cytodex-1, Cytodex-3, Hil-lex® II and Plastic Plus in spinner vessel and two liters bioreactor cultured for 96 hours. Biological performance of the microcarrier in RPMI media showed the preference of Vero cell grew on Cytodex 3 microcarriers with highest maximum viable cell number (2.4 × 105 cells/ml) followed by Cytodex 1, Hillex and Plustic Plus. Vero cell on Cyto-dex-3 data in spinner flask was compared in bioreactor and result showed higher viable cell number in biorector. Thus, this dextran-crosslink gelatin microcarrier (Cytodex 3) provided the best surface for cell attachment and fast proliferation. At the end of this cell growth improvement will be used for virus transfection producing a vaccine in bioreactor.

Highlights

  • Vero cell line was derived from the kidney of a normal, adult, African green monkey (Cercopithecus) [1]

  • The objective of this research was to study the comparison of Vero cell growth on different types of commercial microcarriers; Cytodex-1, Cytodex-3, Hillex® II and Plastic Plus in spinner vessel and two liters bioreactor cultured for 96 hours

  • For Cytodex 1 and Cytodex 3 with each 3 g/l, the microcarriers were swollen with Ca2+, Mg2+-free phosphate buffer saline (PBS) twice and autoclaved at 115oC for 15 minutes while Hillex® (14 g/l) and Plustic Plus (20 g/l) microcarrier were washed with deionized water and autoclaved at 121oC for 30 minutes

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Summary

Introduction

Vero cell line was derived from the kidney of a normal, adult, African green monkey (Cercopithecus) [1]. Vero cells have been extensively used for producing viral vaccines [2,3,4] and for evaluating the performance of animal cell in bioreactors with modified condition [5]. This cell line has been used extensively for virus replication studies and plaque assays [1]. Vero cells are anchorage dependent cells which required solid substrate to attach and grow. Vero cells can be used in microcarrier and suspension cultures for large scale production in bioreactors. Microcarrier culture comprises the cultivation of anchorage dependent cells on small solid substrate suspended in growth medium and can be cultured in larger scale. There are various kinds of microcarriers based on material that were used

Culture Media and Chemicals
Cell Dissociation
Spinner Flask Culture
Bioreactor Culture
Calculation
2.10. Substrate Analysis
Growth Kinetic of Vero Cells in Spinner Flask and Bioreactor Culture
Cell Distributions and Morphologies on MC
Discussions
Conclusions
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