Stem Cells in

Regenerative Medicine

Todd C. McDevitt, Ph.D.

June 21, 2006

The Era of Regenerative Medicine

Tissue Engineering

Regenerative Medicine

(1980s 2005)

(2005+)

Scaffolds +

Stem Cells +

Biomolecules +

Materials (?) +

Cells

Biomolecules (?)

Tissues

Tissues

Objective:

(So whats changed?)

Make the tissue

Repair the tissue

Stem Cells & Tissue Engineering

Somatic cell tissue engineering:
- committed cell types
- tissue assembled upon a
biodegradable scaffold
Stem cell tissue engineering:
- uncommitted cell types
- direct differentiation by
environmental cues

Stock & Vacanti, Annu Rev Med 2001.

Hope or Hype?

Cancer
diagnosis &
treatment

http://stemcells.nih.gov/info/media/DSC_1185.jpg

Stem Cell History

1961 Stem cells first discovered in the blood
1964 Embryonic carcinoma (EC) cell lines derived
1968 First bone marrow transplantation performed
1968 First human egg fertilized in vitro
1981 Mouse embryonic stem (ES) cell lines derived
1984 Human EC cell lines derived
1995 Rhesus monkey ES cell lines derived
1998 Human ES & embryonic germ cell lines derived
2001 Federal approval of human ES cell research
2004 Cloned human ES cells produced

Definition of a Stem Cell

Criteria
1) Self-renewal
- Proliferative
- Clonogenic potential
2) Differentiate into mature
(somatic) cell types
- Often times, multilineage potential

Zandstra & Nagy, Annu Rev Biomed Eng 2001.

Potency of Stem Cells

Modified from:
LaGasse et al., Immunity 2001.

Totipotent all cell/tissue types

Pluripotent embryonic & adult cells
Multipotent multiple cell types

Potential

Stem Cell Potency

Embryonic

Bone marrow

Pluripotent

Multipotent

Satellite cell

Unipotent

Potency affects applications & ease of manipulation

Types of Stem Cells

Embryonic (Pluripotent)
- Stem cells (inner cell mass)
- Germ cells (embryonic gonad)
- Carcinoma (testes)

ES
EG
EC

Isolation of Pluripotent Stem Cells

ES

EG

EC

Donovan & Gearhart, Nature 2001.

Derivation of ES Cells

(Morula)
(Blastocyst)
Adapted from: Gepstein, Circ Res 2002.

Somatic Cell Nuclear Transfer

Lanza et al., Nature Biotech 1999.

Therapeutic cloning
Advanced Cell Technology

Reproductive
cloning

Cloning & Stem Cell Research

A minority of stem cell research involves cloning.

ES Cell Culture & Differentiation

Undifferentiated

Suspension

10x

Outgrowths

Embryoid bodies (EBs)

ES Cell Differentiation

http://www.geron.com/showpage.asp?code=prodst

Types of Stem Cells

Embryonic (Pluripotent)
- Stem cells (inner cell mass)
- Germ cells (embryonic gonad)
- Carcinoma (P19s)

ES
EG
EC

Fetal (Multipotent)
- Umbilical cord, placenta, amniotic fluid

Fetal Stem Cells

- Umbilical cord
- Placenta
- Amniotic fluid

http://webmd.lycos.com/content/asset/adam_imagepage_9980

Types of Stem Cells

Embryonic (Pluripotent)
- Stem cells (inner cell mass)
- Germ cells (embryonic gonad)
- Carcinoma (P19s)

ES
EG
EC

Fetal (Multipotent)
- Umbilical cord, placenta, amniotic fluid
Adult (Multi/Unipotent)
- Bone marrow (hematopoietic & mesenchymal)
- Tissue: neural, skin, skeletal muscle (satellite
cells), fat, endothelial progenitors (EPCs)

Tissue-Derived Adult Stem Cells

Some examples:
Bone marrow
Skin
Neural
- central & peripheral
nervous system

Endothelial progenitors

Even more:
Dental pulp
Cardiac
Pancreas
Liver

- circulating

Adipose tissue
Satellite cells (skeletal muscle)

Intestine

Adult vs. Embryonic Stem Cells

Embryonic

Adult

- Pluripotent
(+)
- Highly proliferative (+)

- Multipotent
- Limited proliferative
capacity

- Non-autologous
- Ethical concerns
- Tumorigenic

- Autologous
(+)
- Less controverserial (+)
- Non-tumorigenic
(+)

Assaying Stem Cell Potential

Developmental
potential assays
1) In vitro differentiation
- Culture dish

2) Teratoma formation
- Grafted into animal

3) Chimera formation
- Injected into blastocyst

Smith, Annu Rev Cell Dev Biol 2001.

Bone Marrow Cell Fate

(-gal, GFP)
Male

Female

Blau, Brazelton & Weiman, Cell 2001.

Bone Marrow Cell Potential

Y chromosome

Gussoni et al.,
Nature 1999.

Dystrophin

FAH+ hepatocytes

Lagasse et al.,
Nat Med 2000.

FAH-/- mouse

mdx mouse

GFP
Cardiomyocyte
Jackson et al.,
J Clin Inv 2001.

Brazelton et al.,
Science 2000.

NeuN

Blau, Brazelton & Weiman, Cell 2001.

Stem Cell Differentiation Models

Frisn, Neuron 2002.

Misinterpretation ?

Frisn, Neuron 2002.

Questions to be Resolved
- What stem cell(s) are useful for which therapies?
Differentiation potential

- How to obtain or derive stem cells?

Cell/tissue source
Genetic manipulation

- What controls stem cell differentiation?

Signals required
Delivery/presentation methods

Stem Cell Translation

Stem Cell Biology

Regenerative Medicine
Clinicians

Identification

Engineers

Cellular
therapies

Derivation

- Quantitative approach
- Environmental control
Propagation

Differentiation
Cell biologists

Tissue
morphogenesis

Adapted from: Gepstein, Circ Res 2002 & http://stemcells.nih.gov/info/media/DSC_1187.jpg