SLIDE 1
Oxygen Biosensor System (OBS)
Islets O2 h
s
O2 O2 O2 O2 O2 O2 O2 O2 O2 O2 O2 O2
O2-sensitive fluorophore 485nm (excite) 615nm (emit)
BD TM Oxygen Biosensor System: Capabilities/Limitations JoAnna - - PowerPoint PPT Presentation
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
SLIDE 2
SLIDE 3
# ## 0.0E+00 2.0E+04 4.0E+04 6.0E+04 8.0E+04 1.0E+05 1.2E+05 1.4E+05 1.6E+05 1.8E+05 24 48 72 96 120 144 168 Time after culture (hours) OCR (fmol/min) 10 25 50 75 100 125 150 200 At 24 hours: y = 801.06x - 4407.5 r2 = 0.9572, P<0.01 At 168 hours: y = 452.67x + 5117 r2 = 0.9447, P<0.01 0.0E+00 2.0E+04 4.0E+04 6.0E+04 8.0E+04 1.0E+05 1.2E+05 1.4E+05 1.6E+05 1.8E+05 50 100 150 200 250 Seeding Density (IE/well) OCR (fmol/min)
A. B.
Effect of Seeding Density (non-
human primate)
Wenjing Wang, et al. Journal Endocrinology (2005) 185, 445-455
SLIDE 4
* * * ## * * * * * # * * * * ** ** 0.0E+00 2.0E+04 4.0E+04 6.0E+04 8.0E+04 1.0E+05 1.2E+05 1.4E+05 1.6E+05 24 48 72 96 120 144 168 Time after culture (hours) OCR (fmol/min) 5.6mM G 16.7mM G 33.3mM G 0.0E+00 2.0E+04 4.0E+04 6.0E+04 8.0E+04 1.0E+05 1.2E+05 1.4E+05 1.6E+05 24 48 72 96 120 144 168 Time after culture (hours) OCR (fmol/min) 5.6mM G 16.7mM G 33.3mM G
B. A.
Effect of [Glucose]
(non-human primate)
Wenjing Wang, et al. Journal Endocrinology (2005) 185, 445-455
SLIDE 5
Effect of [Glucose] (non-human primate)
** ## 0.0E+00 2.0E+04 4.0E+04 6.0E+04 8.0E+04 1.0E+05 1.2E+05 1.4E+05 OCR (fmol/min) 2.8mM G 16.7mM G 2.8mM G again
Wenjing Wang, et al. Journal Endocrinology (2005) 185, 445-455
SLIDE 6
# ** ## ** ** ** ** ## ** ** ** 0.0E+00 2.0E+04 4.0E+04 6.0E+04 8.0E+04 1.0E+05 1.2E+05 1.4E+05 1.6E+05 1.8E+05 30 60 90 120 Incubation Time (minutes) OCR (fmol/min) 5.6mM G 16.7mM G 33.3mM G 0.0E+00 2.0E+04 4.0E+04 6.0E+04 8.0E+04 1.0E+05 1.2E+05 1.4E+05 1.6E+05 1.8E+05 30 60 90 120 Incubation Time (minutes) OCR (fmol/min) 5.6mM G 16.7mM G 33.3mM G
B. A.
Effect of [Glucose]
(human islets)
Wenjing Wang, et al. Journal Endocrinology (2005) 185, 445-455
SLIDE 7
** ** 200 400 600 800 1000 1200 Insulin (ng/ml/100IE) 5.6mM G 16.7mM G 33.3mm G * ** 0.0E+00 1.0E+04 2.0E+04 3.0E+04 4.0E+04 5.0E+04 6.0E+04 7.0E+04 8.0E+04 OCR (fmol/min/100IE) 5.6mM G 16.7mM G 33.3mM G y = 56.178x r
2 = 0.7681, P<0.01
0.0E+00 1.0E+04 2.0E+04 3.0E+04 4.0E+04 5.0E+04 6.0E+04 7.0E+04 8.0E+04 200 400 600 800 1000 1200 Insulin (ng/ml/100IE) OCR (fmol/min/100IE)
C. A. B.
Insulin Secretion and OCR
(human islets)
Wenjing Wang, et al. Journal Endocrinology (2005) 185, 445-455
SLIDE 8
High vs. Low viability islets (human)
* 0.5 1 1.5 2 2.5 3 Stimulation Index of OCR Low viability (<65%) High viability (>80%)
Wenjing Wang, et al. Journal Endocrinology (2005) 185, 445-455
SLIDE 9
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.
SLIDE 10
Current Studies Is the BDTM Oxygen Biosensor System an effective approach for quickly assessing the functional viability of islets prior to transplantation?
SLIDE 11
Acknowledgement
- Dr. Wenjing Wang