BD TM Oxygen Biosensor System: Capabilities/Limitations JoAnna - PowerPoint PPT Presentation

BD TM Oxygen Biosensor System: Capabilities/Limitations JoAnna Reems, PhD Puget Sound Blood Center

Oxygen Biosensor System (OBS) s O 2 O 2 O 2 O 2 O 2 O 2 O 2 h O 2 O 2 O 2 O 2 Islets O 2 -sensitive O 2 O 2 fluorophore 485nm 615nm (excite) (emit)

A. 1.8E+05 10 25 Effect of 50 75 1.6E+05 100 125 150 200 1.4E+05 Seeding OCR (fmol/min) 1.2E+05 1.0E+05 Density (non- ## 8.0E+04 # human primate) 6.0E+04 4.0E+04 2.0E+04 0.0E+00 24 48 72 96 120 144 168 Time after culture (hours) B. 1.8E+05 1.6E+05 1.4E+05 At 24 hours: y = 801.06x - 4407.5 OCR (fmol/min) 1.2E+05 r 2 = 0.9572, P <0.01 1.0E+05 8.0E+04 At 168 hours: y = 452.67x + 5117 6.0E+04 r 2 = 0.9447, P <0.01 4.0E+04 2.0E+04 0.0E+00 0 50 100 150 200 250 Wenjing Wang, et al. Journal Endocrinology Seeding Density (IE/well) (2005) 185, 445-455

A. Effect of 1.6E+05 5.6mM G 16.7mM G 33.3mM G 1.4E+05 ** * [Glucose] 1.2E+05 * OCR (fmol/min) ** * 1.0E+05 (non-human primate) * * * * 8.0E+04 * ## * * * # * 6.0E+04 4.0E+04 2.0E+04 0.0E+00 24 48 72 96 120 144 168 Time after culture (hours) B. 5.6mM G 1.6E+05 16.7mM G 33.3mM G 1.4E+05 1.2E+05 OCR (fmol/min) 1.0E+05 8.0E+04 6.0E+04 4.0E+04 2.0E+04 Wenjing Wang, et al. Journal Endocrinology 0.0E+00 (2005) 185, 445-455 24 48 72 96 120 144 168 Time after culture (hours)

Effect of [Glucose] (non-human primate) 2.8mM G 1.4E+05 16.7mM G 2.8mM G again ** 1.2E+05 1.0E+05 OCR (fmol/min) ## 8.0E+04 6.0E+04 4.0E+04 2.0E+04 0.0E+00 Wenjing Wang, et al. Journal Endocrinology (2005) 185, 445-455

A. Effect of 1.8E+05 5.6mM G 16.7mM G 1.6E+05 33.3mM G ** ## [Glucose] 1.4E+05 ** ## 1.2E+05 ** OCR (fmol/min) (human islets) ** 1.0E+05 ** ** 8.0E+04 6.0E+04 ** # ** 4.0E+04 2.0E+04 0.0E+00 30 60 90 120 Incubation Time (minutes) B. 1.8E+05 5.6mM G 16.7mM G 1.6E+05 33.3mM G 1.4E+05 1.2E+05 OCR (fmol/min) 1.0E+05 8.0E+04 6.0E+04 4.0E+04 2.0E+04 0.0E+00 30 60 90 120 Wenjing Wang, et al. Journal Endocrinology Incubation Time (minutes) (2005) 185, 445-455

A. B. Insulin 8.0E+04 1200 5.6mM G 5.6mM G 16.7mM G 16.7mM G 33.3mm G 33.3mM G ** 7.0E+04 ** Secretion 1000 ** * 6.0E+04 OCR (fmol/min/100IE) 800 Insulin (ng/ml/100IE) and OCR 5.0E+04 4.0E+04 600 (human islets) 3.0E+04 400 2.0E+04 200 1.0E+04 0 0.0E+00 C. 8.0E+04 y = 56.178x 2 = 0.7681, P<0.01 7.0E+04 r 6.0E+04 OCR (fmol/min/100IE) 5.0E+04 4.0E+04 3.0E+04 2.0E+04 1.0E+04 Wenjing Wang, et al. 0.0E+00 Journal Endocrinology 0 200 400 600 800 1000 1200 (2005) 185, 445-455 Insulin (ng/ml/100IE)

High vs. Low viability islets (human) Low viability 3 (<65%) High viability * (>80%) 2.5 Stimulation Index of OCR 2 1.5 1 0.5 0 Wenjing Wang, et al. Journal Endocrinology (2005) 185, 445-455

Summary -Seeding densities correlate with OCR -Glucose stimulated OCRs are measurable with human islets. BD system has the sensitivity to distinguish between islets with high and low viability. -Real-time kinetic measurements readily achievable. -Simultaneous measurements of OCR and insulin secretion.

Current Studies Is the BDTM Oxygen Biosensor System an effective approach for quickly assessing the functional viability of islets prior to transplantation?

Acknowledgement Dr. Wenjing Wang Lisa Upshaw Jondell Clever Yoshiko Tamura Denise Apo Ariel Munsey