Evaluation of Various Methods to Quantify Endothelial Cells Attached to Vascular Prostheses: Comparison with a New “Gold Standard” FACS Method

Visser, M.J.T.; Lennep, A.C.D. van; Bockel, J.H. van; Hinsbergh, Victor W.M. van; Keur, Maarten van der; Hermans, J.

doi:10.1006/jsre.1996.0110

Cited by 1 publication

(1 citation statement)

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“…Cell density and growth were monitored by aseptically sampling the FibraCel matrix periodically. FibraCel disks were incubated in 0.1 M citric acid containing 0.1% (w/v) crystal violet at a density of 5 disks/mL for 2 h at 37 °C with vortexing at 10 min intervals to release the nuclei of FibraCel‐attached cells (15). Cell populations were estimated (16) by counting the released, stained nuclei with a hemocytometer and phase contract microscope (Zeiss).…”

Section: Methodsmentioning

confidence: 99%

Bioreactor Production of Recombinant Herpes Simplex Virus Vectors

Knop

Harrell

2008

Biotechnol Progress

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Serotypical application of herpes simplex virus (HSV) vectors to gene therapy (type 1) and prophylactic vaccines (types 1 and 2) has garnered substantial clinical interest recently. HSV vectors and amplicons have also been employed as helper virus constructs for manufacture of the dependovirus adeno-associated virus (AAV). Large quantities of infectious HSV stocks are requisite for these therapeutic applications, requiring a scalable vector manufacturing and processing platform comprised of unit operations which accommodate the fragility of HSV. In this study, production of a replication deficient rHSV-1 vector bearing the rep and cap genes of AAV-2 (denoted rHSV-rep2/cap2) was investigated. Adaptation of rHSV production from T225 flasks to a packed bed, fed-batch bioreactor permitted an 1100-fold increment in total vector production without a decrease in specific vector yield (pfu/cell). The fed-batch bioreactor system afforded a rHSV-rep2/cap2 vector recovery of 2.8 x 10(12) pfu. The recovered vector was concentrated by tangential flow filtration (TFF), permitting vector stocks to be formulated at greater than 1.5 x 10(9) pfu/mL.

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Section: Methodsmentioning

confidence: 99%