Page 126 - Live-cellanalysis handbook

Live-Cell Analysis Handbook — Third Edition

Microglial phagocytosis of disease-related peptides
Microglia are also responsible for the clearance of other cellular and myelin basic protein. Using iPSC-derived microglia, we
debris and peptides associated with neurological disorders. In observed a concentration-dependent phagocytic clearance of
order to evaluate the engulfment of relevant disease-associated physiologically relevant material (Figure 6). These data support
peptides, the IncuCyte pHrodo labeling kit was used to label the use of pHrodo-labeled material to study microglia function in
®
both apoptotic Neuro-2A, beta-amyloid, alpha-synuclein maintaining brain homeostasis to clear potential toxic factors.

Apoptotic Neuro-2A Beta-Amyloid (1-42)
Orange area x 10 Orange area x 10
5
5
2
(μm /image) (μm /image)
2
2.5 3
33.3 ug/ml
2.0
50 K/well N2A
2 100 ug/ml
1.5

1.0 25 K/well N2A
1 11.1 ug/ml

0.5
12.5 K/well N2A 3.7 ug/ml
6.25 K/well N2A 1.2 ug/ml
Control Vehicle
0.0 0
0 5 10 15 20 25 0 5 10 15 20 25
Time (h) Time (h)
Alpha-Synuclein Myelin basic protein
Orange area x 10 Orange area x 10
5
5
(μm /image) (μm /image)
2
2
3
1.0
300 ug/ml 33.3 ug/ml
2
100 ug/ml
11.1 ug/ml
0.5
1
3.7 ug/ml
33.3 ug/ml
11.1 ug/ml 1.2 ug/ml
3.7 ug/ml 0.4 ug/ml
Vehicle Vehicle
0.0 0
0 5 10 15 20 25 0 5 10 15 20 25
Time (h) Time (h)
Figure 6. Comparison of phagocytic clearance of neuro-associated peptides in an iPSC-derived microglial cell model. iPSC-derived microglia (Axol BioSciences)
seeded at 30,000 cells/well phagocytose physiologically relevant target material. Kinetic graphs display concentration-dependent response to pHrodo labeled
apoptotic Neuro-2A, beta-amyloid, alpha-synuclein and myelin basic protein.

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