| Cell line (human) |
WT iPSC-microglia |
UCI ADRC iPSC Core |
ADRC5;orgin: Blurton-Jones lab |
iPSC-derived microglial line |
| Cell line (human) |
TREM2 KO iPSC microglia |
Blurton-Jones lab |
ADRC5 Clone 28-18;orgin: Blurton-Jones lab |
CRISPR-mediated knockout of TREM2 on the WT iPSC line |
| Cell line (human) |
WT GFP-expressing iPSC-microglia |
Coriell |
AICS-0036;RRID:CVCL_JM19
|
iPSC-line with GFP tagged to αtubulinOriginally developed by Dr. Bruce Conklin |
| Cell line (human) |
TREM2 KO GFP-expressing iPSC-microglia |
Blurton-Jones lab |
GFP Clone 1from above RRID |
CRISPR-mediated knockout of TREM2 on the WT GFP+ iPSC line |
| Cell line (human) |
WT RFP-expressing iPSC-microglia |
Coriell |
AICS-0031-035;RRID:CVCL_LK44
|
iPSC-line with RFP tagged to αtubulinOriginally developed by Dr. Bruce Conklin |
| Cell line (human) |
TREM2 KO RFP-expressing iPSC-microglia |
Blurton-Jones lab |
RFP Clone 6from above RRID |
CRISPR-mediated knockout of TREM2 on the WT RFP+ iPSC line |
| Cell line (human) |
WT Salsa6f-expressing iPSC-microglia |
UCI ADRC iPSC Core |
ADRC76 Clone 8;orgin: Blurton-Jones lab |
iPSC-line expressing a GCaMP6f-tdTomato fusion construct (Salsa6f) |
| Cell line (human) |
TREM2 KO Salsa6f-expressing iPSC microglia |
Blurton-Jones lab |
ADRC76 Clone 8, Clone 98;orgin: Blurton-Jones lab |
CRISPR-mediated knockout of TREM2 on the WT Salsa6f+ iPSC line |
| Cell line (human) |
Orai1 KO iPSC microglia |
Blurton-Jones lab |
ADRC76;orgin: Blurton-Jones lab |
CRISPR-mediated knockout of Orai1 on the WT ADRC76 iPSC line |
| Transfected construct (transgene) |
Salsa6f |
Addgene |
Plasmid# 140188;RRID:Addgene_140188
|
A genetically encoded calcium indicator with tdTomato linked to GCaMP6f by a V5 epitope tag. |
| Other |
DMEM/F12, HEPES, no Phenol red |
Thermo Fisher Scientific |
11038021 |
Microglia differentiation cell culture medium |
| Other |
TeSR-E8 |
STEMCELL Technologies |
05990 |
Stem cell culture medium |
| Other |
StemDiff Hematopoietic kit |
STEMCELL Technologies |
05310 |
|
| Peptide, recombinant protein |
Nonessential amino acids |
Gibco |
11140035 |
|
| Peptide, recombinant protein |
GlutaMAX |
Gibco |
35050061 |
|
| Peptide, recombinant protein (human) |
Insulin |
Sigma |
I2643 |
|
| Peptide, recombinant protein |
B27 |
Gibco |
17504044 |
|
| Peptide, recombinant protein |
N2 |
Gibco |
A1370701 |
|
| Peptide, recombinant protein |
Insulin-transferrin-selenite |
Gibco |
41400045 |
|
| Peptide, recombinant protein |
IL-34 |
PeproTech |
200-34 |
|
| Peptide, recombinant protein |
TGFβ1 |
PeproTech |
100-21 |
|
| Peptide, recombinant protein |
M-CSF |
PeproTech |
300-25 |
|
| Peptide, recombinant protein |
CX3CL1 |
PeproTech |
300-31 |
|
| Peptide, recombinant protein |
CD200 |
Novoprotein |
C311 |
|
| Peptide, recombinant protein |
Fibronectin |
STEMCELL Technologies |
07159 |
|
| Other |
Matrigel |
Corning |
356231 |
|
| Other |
ReLeSR |
STEMCELL Technologies |
5872 |
Human pluripotent stem cell selection and passing reagent |
| Other |
Goat serum |
Thermo Fisher Scientific |
10,000C |
|
| Other |
Fluorescent beta-amyloid 1–42 (647) |
AnaSpec |
AS64161 |
|
| Other |
pHrodo tagged zymosan A beads |
Thermo Fisher Scientific |
P35364 |
|
| Other |
pHrodo tagged S. aureus
|
Thermo Fisher Scientific |
A10010 |
|
| Other |
Human Stem Cell Nucleofector kit 2 |
Lonza |
VPH-5022 |
|
| Other |
Alt-R CRISPR-Cas9 tracrRNA |
IDTDNA |
107253 |
|
| Other |
Alt-R HiFi Cas9 Nuclease |
IDTDNA |
1081061 |
|
| Antibody |
Anti-human IBA1(rabbit monoclonal) |
Wako |
019-19741;RRID:AB_839504
|
(1:200) |
| Antibody |
Goat anti-rabbit 555(secondary antibody) |
Thermo Fisher Scientific |
A21429;RRID:AB_2535850
|
(1:400) |
| Other |
Human TruStain FcX |
BioLegend |
Cat# 422301 |
Fc blocking solution5 μL per test |
| Antibody |
Brilliant Violet 421 anti-human P2RY12Primary antibody(mouse monoclonal) |
BioLegend |
392105;clone 16001E;RRID:AB_2783290
|
(5 μL) per test |
| Antibody |
Brilliant Violet 421 mouse IgG2aκ Isotype control mouse |
BioLegend |
407117;clone MOPC-173;RRID:AB_2687343
|
(5 μL) per test |
| Chemical compound, drug |
Fluo-4 AM |
Thermo Fisher Scientific |
F14201 |
|
| Chemical compound, drug |
Fura-red AM |
Thermo Fisher Scientific |
F3021 |
|
| Chemical compound, drug |
Pluronic F-127 |
Thermo Fisher Scientific |
P3000MP |
|
| Chemical compound, drug |
Cal-520 AM |
AAT Bioquest |
21130 |
|
| Chemical compound, drug |
Cal-590 AM |
AAT Bioquest |
20510 |
|
| Chemical compound, drug |
ci-IP3/PM |
SiChem |
6210 |
Caged-inositol triphosphate analog |
| Chemical compound, drug |
Hoeschst |
Thermo Fisher Scientific |
R37165 |
|
| Chemical compound, drug |
ADP |
Sigma-Aldrich |
A2754 |
|
| Chemical compound, drug |
ATP |
Sigma-Aldrich |
A9187 |
|
| Chemical compound, drug |
UTP |
Sigma-Aldrich |
U1006 |
|
| Chemical compound, drug |
PSB 0739 |
Tocris |
3983 |
|
| Chemical compound, drug |
MRS 2211 |
Tocris |
2402 |
|
| Chemical compound, drug |
Synta66 |
Sigma-Aldrich |
SML1949 |
Orai channel inhibitor |
| Chemical compound, drug |
2-APB |
Sigma-Aldrich |
D9754 |
|
| Chemical compound, drug |
Gadolinium |
Sigma-Aldrich |
G7532 |
|
| Chemical compound, drug |
EGTA |
Sigma-Aldrich |
E8145 |
|
| Chemical compound, drug |
1-Thioglycerol |
Sigma-Aldrich |
M6145 |
|
| Chemical compound, drug |
CloneR |
STEMCELL Technologies |
05888 |
Defined supplement for single-cell cloning of human iPS cells |
| Chemical compound, drug |
Thiazovivin |
STEMCELL Technologies |
72252 |
ROCK inhibitor |
| Other |
35 mm glass-bottom dish |
MatTek |
P35G-1.5-14C |
1.5 coverslip, 14 mm glass diameter |
| Other |
Incubation perfusion Lid for 35 mm dishes |
Tokai Hit |
LV200-D35FME |
Perfusion lid with inlet and outlet |
| Other |
Laser Scanning Confocal Microscope |
Olympus |
FV3000 |
Equipped with Resonant Scanner, IX3-ZDC2 Z-drift compensator, 40× silicone oil objective, 20× air objective |
| Other |
Stage Top Incubation System |
Tokai Hit |
STXG |
Temperature and humidity control for FV3000 microscope stage |
| Other |
Nikon Eclipse Ti microscope system |
Nikon |
|
Equipped with a 40× oil immersion objective (NA 1.3; Nikon) and an Orca Flash 4.0LT CMOS camera (Hamamatsu) |
| Other |
Chemotaxis Assay Chamber |
Hansang Cho Lab |
|
|
| Other |
IncuCyte S3 Live-Cell Analysis System |
Sartorius |
|
|
| Other |
Essen Incucyte WoundMaker |
Sartorius |
4493 |
|
| Software, algorithm |
GraphPad Prism 9.1.0 |
|
|
Data analysis, statistical analysis |
| Software, algorithm |
Fiji (ImageJ) |
|
|
Image analysis |
| Software, algorithm |
Incucyte 2020C |
|
|
Image acquisition and analysis |
| Software, algorithm |
Imaris 9.7.0 |
|
|
Cell tracking and image analysis |
| Software, algorithm |
Flika |
|
|
Image analysis |
| Software, algorithm |
DiPer Excel Macros |
|
PMID:25033209
|
Data analysis, directional persistence |
