www.bumc.bu.edu/stemcellswww.bumc.bu.edu/stemcells

Advancing Regenerative Medicine withThe Boston University�Induced Pluripotent Stem (iPS) Cell Bank

Darrell N. Kotton, MDGustavo Mostoslavsky, MD, PhD

George Murphy, PhDWellington Cardoso, MD, PhD

Joyce Wong, PhDFeb. 16, 2010

iPS Cell Bank

• What is it?

• Why is it important?

• How far off are we from using the Bank for Regenerative Medicine applications?

Embryonic Stem Cells Embryonic Stem Cells -- ESCESC

BlastocystBlastocyst

ES cell colonyES cell colony

55--6 d6 daysays

Fertilized eggFertilized egg

EndodermEndodermEndoderm

MesodermMesodermMesoderm

EctodermEctodermEctoderm

ESC Major Limitations for Regenerative MedicineESC Major Limitations for Regenerative Medicine

ESCESC iPS cellsiPS cells

Technical hurdlesTechnical hurdles

Ethical concernsEthical concerns

ImmunogenicityImmunogenicity

ESCESC

Technical hurdlesTechnical hurdles

Ethical concernsEthical concerns

ImmunogenicityImmunogenicity

‘Reprogramming’

ESCell

PluripotentDifferentiatedSkin Fibroblasts

iPSCell

Takahashi and Yamanaka.Cell 2006.

The Discovery of Induced Pluripotent Stem (iPS) Cells

Oct4Sox2Klf4

cMyc

‘Reprogramming’ with STEMCCA

55’’ LTRLTR PSIPSI RRERREHIV cpPuHIV cpPu

Oct4Oct4 F2AF2A Klf4Klf4 IRESIRES Sox2Sox2 E2AE2A cMyccMyc

WPREWPREdU3dU3

33’’ LTRLTR

STEMCCASTEMCCA

EF1EF1 or TetOor TetO

Tail Fibroblast

iPSCell

iPS derivation without the presence of exogenous transgenesiPS derivation without the presence of exogenous transgenes

55’’ LTRLTR PSIPSI RRERREHIV cpPuHIV cpPu

Oct4Oct4 F2AF2A Klf4Klf4 IRESIRES Sox2Sox2 E2AE2A cMyccMyc

WPREWPREdU3dU3

33’’ LTRLTR

STEMCCASTEMCCA

EF1EF1 or TetOor TetO

loxPloxP

STEMCCASTEMCCA

EF1EF1

loxploxp

55’’ LTRLTR PSIPSI RRERREHIV cpPuHIV cpPu

Oct4Oct4 F2AF2A Klf4Klf4 IRESIRES Sox2Sox2 E2AE2A cMyccMycWPREWPRE

dU3dU333’’ LTRLTR

loxploxpSTEMCCASTEMCCA

55’’ LTRLTR PSIPSI RRERREHIV cpPuHIV cpPu

EF1EF1

Oct4Oct4 F2AF2A Klf4Klf4 IRESIRES Sox2Sox2 E2AE2A

WPREWPREdU3dU3

33’’ LTRLTR

loxploxp

dU3dU3

ChromChrom ChromChrom

dU3dU3

ChromChrom

loxploxpChromChrom

Reverse Transcription Reverse Transcription + integration + integration

Cre excisionCre excision

cMyccMyc

Boston University iPS Cell Bank

• What is it?- A repository of human iPS cells generated

from individuals affected by diseases treated or studied at BUSM/BMC

•Why is it important?

The Potential of iPS Research The Potential of iPS Research

•• Skin fibroblastsSkin fibroblasts•• Blood cellsBlood cells•• Hair keratinocytesHair keratinocytes•• ????

iPS cellsiPS cells

cell therapycell therapy

Normal embryo developmentNormal embryo developmentDisease modelsDisease modelsDrug screeningDrug screening

Differentiation into

Differentiation into

specific lineages

specific lineages

Emphysema (AAT Def.)Cystic Fibrosis Diabetes

IBD Sickle Cell AmyloidosisLong QT Syndrome

Transduction/Transduction/TransfectionTransfection

Nucle

ar Re

progra

mming

Nucle

ar Re

progra

mming Oct4

Klf4 Sox2

cMyc

Boston University iPS Cell Bank

• What is it?- A repository of human iPS cells generated

from individuals affected by diseases treated or studied at BUSM/BMC

•Why is it important?-Model disease/development-Develop cell-based therapies

•How far off are we from using the Bank for Regenerative Medicine applications?

ARC StructureSkin biopsyFibroblasts

Reprogram

STEMCCA

iPS Cell Bank

EndodermLungLiverGutPancreas

Kotton, Mostoslavsky,Cardoso

MesodermBloodEndotheliumCardiomyocytesOther mesenchyme

Murphy, MostoslavskyWong, Keller

In vivo DeliveryScaffoldsBiocapsules

BU BMEWong

Tra1-81

Cystic Fibrosis

JC JeanAba SomersChris FordAmel Omari

Emphysema(AAT Deficiency)

Scleroderma

Long QT Syndrome

Disease-Specific Human iPS Cell LinesAmyloid (TTR)

Sickle Cell Dz

Brenden SmithGeorge MurphyCesar SommerBob Lafyatis

Regenerative Medicine ARCDarrell Kotton Lab

George Murphy Lab

www.bumc.bu.edu/stemcellswww.bumc.bu.edu/stemcells

Joyce Wong LabWellington Cardoso Lab

Gustavo Mostoslavsky Lab