Pluripotent Stem Cell CharacterizationA guide for building a complete characterization profile for PSCs

Profiling step Product Cat. No.

Check simple indicators Alkaline Phosphatase Live Stain A14353

Confirm surface markers Alexa Fluor® 488 CD44 A25528

Alexa Fluor® 488 Tra-1-60 A25618

Alexa Fluor® 555 Tra-1-60 A24879

Alexa Fluor® 594 Tra-1-60 A24882

Perform flow cytometry SSEA4 Mouse Anti-Human mAb (clone eBioMC-813-70 (MC-813-70)), Alexa Fluor® 488 Conjugate

A14810

SSEA4 Mouse Anti-Human mAb (clone MC813-70), Alexa Fluor® 647 Conjugate SSEA421

SSEA4 Mouse Anti-Human mAb (clone eBioMC-813-70 (MC-813-70)), PE Conjugate A14766

CD44 Rat Anti-Human/Mouse mAb (clone IM7) Fluorescein (FITC) Conjugate A16193

CD44 Rat Anti-Mouse mAb (clone IM7.8.1), PE Conjugate RM5704

Confirm nuclear markers Pluripotent Stem Cell 4-Marker Immunocytochemistry Kit A24881

Pluripotent Stem Cell Immunocytochemistry Kit (Sox2, Tra-1-60) A25525

Pluripotent Stem Cell Immunocytochemistry Kit (Oct4, SSEA4) A25526

Detect germ layer markers 3-Germ Layer Immunocytochemistry Kit A25538

Quantify trilineage potential TaqMan® hPSC Scorecard™ Panel, 96-well fast (1 kit) A15871

TaqMan® hPSC Scorecard™ Panel, 96-well fast (1 panel) A15876

TaqMan® hPSC Scorecard™ Panel, 384-well (1 kit) A15872

TaqMan® hPSC Scorecard™ Panel, 384-well (1 panel) A15870

Analyze differentiation Human Neural Stem Cell Immunocytochemistry Kit A24354

Human Cardiomyocyte Immunocytochemistry Kit A25973

Ordering information

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Let us do it for you.

With access to a complete portfolio of characterization tools and cutting-edge instruments, the CellModel™ Services team can perform numerous stem cell characterization experiments using cell lines that you provide or ones that we customize for you.

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For Research Use Only. Not for use in diagnostic procedures. © 2014 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. TaqMan is a registered trademark of Roche Molecular Systems, Inc., used under permission and license. CO010177 0714

Abbreviations: ACTN2: alpha actinin 2; AFP: alpha fetoprotein; AP: alkaline phosphatase; CD44: cluster of differentiation 44; DAPI: 4’,6-diamidino-2-phenylindole; iPSC: induced pluripotent stem cell; MYH6: myosin, heavy chain 6; NKX2.5: NK2 homeobox 5; PAX6: paired box gene 6; OCT4: octamer-binding transcription factor 4; SMA: smooth muscle actin; SOX1/SOX2: sex-determining region Y, box 1 or 2; SSEA: stage-specific embryonic antigen; TNNT2: troponin T type 2; TRA: terato-related antigen; TUBB3/TUJ1: beta-III tubulin

To characterize cardiomyocytes: ACTN2, MHY6, NKX2.5, TNNT2 OCT4–+

To characterize NSCs: Nestin, PAX6, SOX1, SOX2 OCT4–+

Differentiated cells

To characterize germ layer specification: AFP (endoderm), SMA (mesoderm), TUJ1 (ectoderm)+

To characterize undifferentiated hPSCs: AP, Nanog, OCT4, SOX2, SSEA4, TRA-1-60, TRA-1-81

CD44, SSEA1–+

Pluripotent stem cellsChoose the right markers for the job

What markers work best for you?

I CAN DIFFERENTIATE

!

» ANALYZE DIFFERENTIATION

Cardiomyocytes

NSCs

DIFFERENTIATE

Karyotype <10% abnormalKaryotype <10% abnormal

» CHECK GENETIC STABILITY

>10passages

bank →I A

M PLURIPOTENT.

» QUANTIFY TRILINEAGE POTENTIAL

FAST

ER THAN TERATOMAS

TaqMan ® hPSC Scorecard™ produc

ts

+ ++

Robust software

Compare93 genes

Robust software

Compare93 genes

» DE

TECT

GERM

LAYE

R MAR

KERS

ENDODERM

MESODERM

ECTODERM

AFP

SMA

TUJ1

» CON

FIRM NU

CLEA

R MAR

KERSSSEA4 OCT4 DAPI

= merge

Counterstain+ Cell

nucleusmarker

+ Cellsurfacemarker

DAPIDAPI

OCT4OCT4SSEA4SSEA4

Are fixed cells pluripotent?

+ +

Improve accuracy with multiple markers

Check culture quality» PERFORM FLOW CYTOMETRY

EXPA

ND

» CONFIRM SURFACE MARKERS

- Cell surfacemarker

+ Cell surface marker

TRA-1-60 CD44CD44

Are live cells pluripotent?

+

CD44

Tra-1-60

= merge

Increase confidence for predicting colony success

» CHECK SIMPLE INDICATORS

Enzyme indicator(alkaline phosphatase)

Are live cellspluripotent?

AP

AP Live Stain

Quickly check colonies prior to selection

SELECT

??AM I P

LURIPOTENT?

Be confident! A comprehensive characterization profile verifies the integrity of your iPSC line. Using a combination of cellular and molecular methods at multiple checkpoints affords maximum confidence in PSC colony selection and utility downstream. The graphic below presents a guide (from left to right) of recommended approaches to complete your characterization profile.

I SEEM PLURIPOTENT...

Creating a complete characterization profile for new iPSC lines

CULTUREENGINEERINGDIFFERENTIATIONCHARACTERIZATION

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