Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving viable micro-organism
Patent
1990-11-20
1993-03-16
Lacey, David L.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving viable micro-organism
4352402, 43524023, 435240241, 435 39, C12Q 106, C12Q 104, C12N 500, C12N 508
Patent
active
051943735
ABSTRACT:
Determination of effectiveness of microvascular endothelial cell seeding upon a vascular graft surface within the operating room environment would be desirable to maintain quality control in any clinical trial. A number of fluorescent dyes including mithramycin, Hoechst 33342, sulfofluorescein diacetate, Nile Red, rhodamine 123, and PKH26-GL were evaluated for their ability to fluorescently label uncultured microvascular endothilial cells on graft material and subsequently allow determination of seeded cell number and cell spreading. Rhodamine 123 and PKH26-GL produced the most desirable characteristics. The selected non-toxic fluorescent dyes allowed excellent cell visualization after a 30 minute incubation. Unlike the other fluorescent dyes evaluated, the selected non-toxic fluorescent dyes caused the cellular cytoplasm to fluoresce bright orange at a 510 nm excitation wavelength while the underlying polyethyleneterephthalate polyester or expanded polytetrafluorethylene demonstrated minimal autofluorescence. No inhibitory effect on cell attachment to plastic or subsequent cell growth in culture was observed. This technique is useful in the operating room to visualize part or all of an microvascular endothelial cell-seeded graft and to permit a quantitative as well as qualitative evaluation of the seeding process to enhance graft patency.
REFERENCES:
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Jarrell Bruce E.
Williams Stuart K.
Elliott George C.
Lacey David L.
Thomas Jefferson University
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