Page 152 - Live-cellanalysis handbook

Live-Cell Analysis Handbook — Third Edition

Validated cell lines
NOTE: Detailed protocols are available for some of the validated cell lines by clicking on the cell type name.

CELL CHEMOATTRACTANT (CONCENTRATION
COATING SETTING RANGE TESTED)
THERAPEUTIC (20 ΜL TOP/ SEED DENSITY TIME
AREA CELL TYPE ASSAY MEDIUM 150 ΜL BOTTOM (CELLS/WELL) (MIN) nM ng/mL
5 μg/mL Fibronectin
+ 0.1% BSA in D-PBS
Jurkat RPMI + 0.5% FBS 50 μg/mL Matrigel 5,000 45-60 SDF-1a (12. 5 – 125) SDF-1a (100– 1,000)
+ 10% FBS in assay
medium (for clustered
cell migration)
20 μg/mL Protein G SDF-1a (3.1 – 200) SDF-1a (24.8 – 1,600)
T Cells RPMI + 0.5% FBS followed by 5 μg/mL 5,000 45-60 CXCL11 (12.5 – 800) CXCL11 (103.8 – 6640)
ICAM RANTES (16) RANTES (125)
C5a (2.6 – 1,900) C5a (21.6 – 15,770)
50 μg/mL Matrigel
Non-adherent IMMUNOLOGY Neutrophils RPMI + 0.5% HSA + 10% FBS in assay 5,000 45-60 LTB4 (1.4 – 1,000) IL-8 (13.4 – 10,000)
fMLP (1.4 – 1,000)
fMLP (0.6 – 437.6)
IL-8 (1.5 – 1120)
medium
LTB4 (0.5 – 336.5)
20 μg/mL Protein G
CCL22 (1 – 100)
CCL22 (8.1 – 810)
RPMI + 50%
followed by 5 μg/
CCRF-CEM
Human Serum mL ICAM OR 5 μg/mL 5,000 45-60 SDF-1a (1 – 100) SDF-1a (8 – 800 nM)
Fibronectin
50 μg/mL Matrigel RANTES (08 – 780) RANTES (0.1 – 100)
RPMI + 0.5%
THP-1 + 10% (dialyzed FBS in 5,000 45-60 MIP-1a (0.8 – 780) MIP-1a (0.1 – 100)
dialyzed FBS
assay medium) MCP-1 (12 – 8,600) MCP-1 (1.4 – 1,000)
Differentiated RPMI + 0.5% FBS 5 μg/mL Fibronectin 2,500 **45-60 C5a (0.5 – 2,000) C5a (4.2 – 16,600)
THP-1 in D-PBS
50 μg/mL Matrigel
Primary
Macrophages RPMI + 0.5% FBS + 10% FBS in assay 2,000 15-30 C5a (10 – 1000) C5a (83 – 8,300)
medium
EBM-2 + ITS + 5 μg/mL Fibronectin 1,000 15 VEGF (0.01 – 2.6)bFGF VEGF (0.4 – 100)
VASCULAR #supplements 10 μg/mL Fibronectin bFGF (0.06 – 5.8) FBS (0.4 – 10%)
0.25% FBS With
(0.02 – 17.4)FBS (0.4
bFGF (0.4 – 300)
HUVEC
+ 0.1% BSA in D-PB
– 10%)
EBM-2 + 0.5%
bFGF (0.01-0.1)
FBS + ITS with
HMVEC
#supplements + 0.1% BSA in D-PB 1,000 15 FBS (10%) FBS (10%)
Adherent MDA-MB-231 DMEM + 2.5 FBS None 1,000 15 FBS (10%) – slight FBS (10%) – slight
F12 + ITS*
FBS (0.01 – 10)%
HT 1080
EGF (0.2 – 1.6)
EGF (1 – 10n)
ONCOLOGY DMEM/F12 response response
1,000
None
15
hydrocortozone +
MCF10a + 10 μg/mL insulin 5 μg/mL Collagen IV in 1,000 15 EGF (0.2 – 17,200) EGF (0.1 – 1,000)
0.05M HCl
100ng/mL cholera
toxin
* Insulin-Transferrin-Selenium (Life Technologies Cat# 41400-045)
# Supplements: EGM-2 singlequot bullet kit (LONZA CC-4176) with growth factors removed and reduced FBS
** Cell settling time pre-differentiation

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