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Non-Invasive Optical Sensor Based Approaches for Monitoring Virus Culture to Minimize BSL3 Laboratory Entry.

Ragupathy V, Setty MK, Kostov Y, Ge X, Uplekar S, Hewlett I, Rao G - Sensors (Basel) (2015)

Bottom Line: Routine PPE use involves significant recurring costs.Alternative non-invasive optical sensor based approaches to remotely monitor cell culture may provide a promising and cost effective approach to monitor infectious virus cultures resulting in lower disruption and costs.The replacement of culture media for cell and virus propagation and virus load monitoring was effectively performed using this fluorescent sensor and resulted in half the number of visits to the BSL3 lab (five versus ten).

View Article: PubMed Central - PubMed

Affiliation: LMV/DETTD/OBRR/CBER/FDA, Silver Spring, MD 20993, USA. viswanath.ragupathy@fda.hhs.gov.

ABSTRACT
High titers of infectious viruses for vaccine and diagnostic reference panel development are made by infecting susceptible mammalian cells. Laboratory procedures are strictly performed in a Bio-Safety Level-3 (BSL3) laboratory and each entry and exit involves the use of  disposable Personnel Protective Equipment (PPE) to observe cell culture conditions. Routine PPE use involves significant recurring costs. Alternative non-invasive optical sensor based approaches to remotely monitor cell culture may provide a promising and cost effective approach to monitor infectious virus cultures resulting in lower disruption and costs. We report here the monitoring of high titer cultures of Human Immunodeficiency Virus-1 (HIV-1) and Herpes Simplex Virus-2 (HSV-2) remotely with the use of optical oxygen sensors aseptically placed inside the cell culture vessel. The replacement of culture media for cell and virus propagation and virus load monitoring was effectively performed using this fluorescent sensor and resulted in half the number of visits to the BSL3 lab (five versus ten).

No MeSH data available.


Related in: MedlinePlus

HIV culture monitoring through oxygen sensor and correlation with HIV-1 viral load (p24 ng/mL) quantification. Each spike in the figure indicates a BSL-3 visit was made to replenish cells and collect an aliquot of culture supernatant for HIV titer determination.
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sensors-15-14864-f001: HIV culture monitoring through oxygen sensor and correlation with HIV-1 viral load (p24 ng/mL) quantification. Each spike in the figure indicates a BSL-3 visit was made to replenish cells and collect an aliquot of culture supernatant for HIV titer determination.

Mentions: In our present work, potential respiration differences between flasks containing infected and un-infected cultures were compared in terms of continuous monitoring using the fluorescent dye patch. The aim was to evaluate whether this approach could help determine when to feed the flask with cells or subculture them. CEM-ss cells were infected as described in methods and the dissolved oxygen in the flask was continuously monitored for ~400 h (Figure 1). Sensor sampling was performed every 30 min.


Non-Invasive Optical Sensor Based Approaches for Monitoring Virus Culture to Minimize BSL3 Laboratory Entry.

Ragupathy V, Setty MK, Kostov Y, Ge X, Uplekar S, Hewlett I, Rao G - Sensors (Basel) (2015)

HIV culture monitoring through oxygen sensor and correlation with HIV-1 viral load (p24 ng/mL) quantification. Each spike in the figure indicates a BSL-3 visit was made to replenish cells and collect an aliquot of culture supernatant for HIV titer determination.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4541811&req=5

sensors-15-14864-f001: HIV culture monitoring through oxygen sensor and correlation with HIV-1 viral load (p24 ng/mL) quantification. Each spike in the figure indicates a BSL-3 visit was made to replenish cells and collect an aliquot of culture supernatant for HIV titer determination.
Mentions: In our present work, potential respiration differences between flasks containing infected and un-infected cultures were compared in terms of continuous monitoring using the fluorescent dye patch. The aim was to evaluate whether this approach could help determine when to feed the flask with cells or subculture them. CEM-ss cells were infected as described in methods and the dissolved oxygen in the flask was continuously monitored for ~400 h (Figure 1). Sensor sampling was performed every 30 min.

Bottom Line: Routine PPE use involves significant recurring costs.Alternative non-invasive optical sensor based approaches to remotely monitor cell culture may provide a promising and cost effective approach to monitor infectious virus cultures resulting in lower disruption and costs.The replacement of culture media for cell and virus propagation and virus load monitoring was effectively performed using this fluorescent sensor and resulted in half the number of visits to the BSL3 lab (five versus ten).

View Article: PubMed Central - PubMed

Affiliation: LMV/DETTD/OBRR/CBER/FDA, Silver Spring, MD 20993, USA. viswanath.ragupathy@fda.hhs.gov.

ABSTRACT
High titers of infectious viruses for vaccine and diagnostic reference panel development are made by infecting susceptible mammalian cells. Laboratory procedures are strictly performed in a Bio-Safety Level-3 (BSL3) laboratory and each entry and exit involves the use of  disposable Personnel Protective Equipment (PPE) to observe cell culture conditions. Routine PPE use involves significant recurring costs. Alternative non-invasive optical sensor based approaches to remotely monitor cell culture may provide a promising and cost effective approach to monitor infectious virus cultures resulting in lower disruption and costs. We report here the monitoring of high titer cultures of Human Immunodeficiency Virus-1 (HIV-1) and Herpes Simplex Virus-2 (HSV-2) remotely with the use of optical oxygen sensors aseptically placed inside the cell culture vessel. The replacement of culture media for cell and virus propagation and virus load monitoring was effectively performed using this fluorescent sensor and resulted in half the number of visits to the BSL3 lab (five versus ten).

No MeSH data available.


Related in: MedlinePlus