Cell Culture, Technology: Enhancing the Culture of Diagnosing Human Diseases

Abstract

Cell culture involves a complex of processes of cell isolation from their natural environment (in vivo) and subsequent growth in a controlled environmental artificial condition (in vitro). Cells from specific tissues or organs are cultured as short term or established cell lines which are widely used for research and diagnosis, most specially in the aspect of viral infection, because pathogenic viral isolation depends on the availability of permissible cell cultures. Cell culture provides the required setting for the detection and identification of numerous pathogens of humans, which is achieved via virus isolation in the cell culture as the "gold standard" for virus discovery. In this review, we summarized the views of researchers on the current role of cell culture technology in the diagnosis of human diseases. The technological advancement of recent years, starting with monoclonal antibody development to molecular techniques, provides an important approach for detecting presence of viral infection. They are also used as a baseline for establishing rapid tests for newly discovered pathogens. A combination of virus isolation in cell culture and molecular methods is still critical in identifying viruses that were previously unrecognized. Therefore, cell culture should be considered as a fundamental procedure in identifying suspected infectious viral agent.

Keywords: Pathogen discovery; Recombinant protein; Transgenic cell line; Viral isolation.

[Table/Fig-1]:

a) Visible virus induced cytopathic effect in DH82 cells 8 days post infection with Bunya virus. The infected cells show visible granular particles and differentiated into macrophages with elongated pseudopodia. b) Bunya virus grown in vero cells, detected on immunofluorescenceassay [31].

[Table/Fig-2]:

a) Negative stain Bunya virus purified from infected Vero cells. b) Transmission electron microscopy of virus infected cells (DH82) shown by black arrows [31].

[Table/Fig-3]:

Result of successful positive recombinant colonies ligation. Lane 1: DNA ladder 1kb; Lane 2-13: positive colonies with NS1; Lane 4, 7, 8 and 12: negative colonies with no NS1 inserted [53].

[Table/Fig-4]:

SDS-PAGE profile of un-purified expressed NS1 (13KDa) protein and un-induced plasmid. Lane 1: protein ladder; Lane 2 - 4: un-induced recombinant plasmid at zero hour; Lane 5 and 6: un-purified recombinant NS1 (13KDa) protein [54].

[Table/Fig-5]:

Result of western blot analysis of the purified proteins from E. coli containing pRSET B/NS1. Lane 1: protein ladder; Lane 2 and 3: purified recombinant NS1 protein [54].