Umbilicalcord

Fetal liverAdult bone marrow

Absolute No. of stem cells and quiescent progenitors

Yolk Sac

Periodo mesoblastico(sacco vitellino)

Periodo epatico (6a settimana)

SpleenPeriodo epato-splenico

(12a settimana)

Periodo midollare (20a settimana)

MICROAMBIENTE MIDOLLARE

Componentecellulare

Componente vascolareCellule reticolariStruttura nervosaAdipociti, fibroblasti, MacrofagiCellule stam. mesenchimali

Matriceextracellulare

Fibre reticolari/collageneProlungamenti cell. reticolariProteine d’adesioneproteoglicani

MIDOLLO OSSEO

IMMISSIONE IN CIRCOLO DEGLI ELEMENTI MATURI

SANGUE PERIFERICO

ERITROCITIPIASTRINE

GRANULOCITINEUTROFILI

GRANULOCITIEOSINOFILI

GRANULOCITIBASOFILI

MONOCITI LINFOCITI

SANGUE PERIFERICO

EMIVITA DELLE CELLULE DEL SANGUE

ERITROCITI 120 giorni

PIASTRINE 4-5 giorni

GRANULOCITI 6-8 ore

MONOCITI 8 ore

LINFOCITI VARIABILE

HEMATOPOIESIS

Stem cell

SELFRENEWAL

CFU-GEMM

STEM CELL POOL

CFU-GM

CFU-Eo

CFU-Bas

BFU-E

CFU-Mk

CFU-G

CFU-MMonocytes

Erytrocytes

CFU-E

Platelets

PROLIFERATIVEPOOL

MATURATIVEPOOL

MATUREBLOOD CELLS

Megakaryocytes

Neutrophyls

Basophyls

Eosinophyls

LTC-ICELTC-IC

MYELOPOIESIS

CYTOKINES REGULATION OF HAEMOPOIESIS

Fattori trascrizionali emopoietici

RARs PU.1 C/EBPε

Sp1

C/EBPs

Egr-1 Rb p53

PU.1 C/EBPs

MICSBP

PU.1

GMP

PU.1C/ EBPs

PU.1

GATA-1

CMP

MEP

GATA -1

BasoGATAs

MITF

Meg

GATA-1,GATA-2NF-E2,RUNX1,FLI1

E

GATA-1,LMO2

B

PU.1

Pax 5

T

GATA-3

CLP

Ikaros

HSC

CBFβ,HOXA9,CUTL-1

SCL/LMO2, ID1

GATA-2,Ikaros

G

C/EBPαPU.1

EoP

C/EBPβ

GATA-1

+/-C/EBPs

Sp1GATA-1;PU.1

Hoxa5

PU.1

Hoxa10

TAL-1,LDB1

E2A,E47

Normal hematopoiesis

CFU-GM

CFU-M

CFU-G

CFU-MEG

Limphoid progenitor

Quiescent Staminal cell

Myeloblast

Monoblast

Megakaryoblast

Pro-erythroblast Erythroblast

Granulocyte

Monocyte

Platelets

Red cells

CFU-GEMM

BFU-E / CFU-E

Epo

SCF, TPO, IL3, GM-CSF

SCF, IL6 IL3, GM-CSF

Plasma cellB lymphocyte

Proliferating staminal cell

SCF, IL1, IL3, IL6, Flt3

ligand, IL11

TGFβ, TNFα, INFγ, LIF

SELF RENEWAL COMMITTMENT PRECURSOR EXPANSION TERMINAL DIFFERENTIATION

Normal hematopoiesis

CFU-GM

CFU-M

CFU-G

CFU-MEG

Limphoid progenitor

Quiescent Staminal cell

Myeloblast

Monoblast

Megakaryoblast

Pro-erythroblast Erythroblast

Granulocyte

Monocyte

Platelets

Red cells

CFU-GEMM

BFU-E / CFU-E

Epo

SCF, TPO, IL3, GM-CSF

SCF, IL6 IL3, GM-CSF

Plasma cellB lymphocyte

Proliferating staminal cell

SCF, IL1, IL3, IL6, Flt3

ligand, IL11

TGFβ, TNFα, INFγ, LIF

SELF RENEWAL COMMITTMENT PRECURSOR EXPANSION TERMINAL DIFFERENTIATION

Normal hematopoiesis

CFU-GM

CFU-M

CFU-G

CFU-MEG

Limphoid progenitor

Quiescent Staminal cell

Myeloblast

Monoblast

Megakaryoblast

Pro-erythroblast Erythroblast

Granulocyte

Monocyte

Platelets

Red cells

CFU-GEMM

BFU-E / CFU-E

Epo

SCF, TPO, IL3, GM-CSF

SCF, IL6 IL3, GM-CSF

Plasma cellB lymphocyte

Proliferating staminal cell

SCF, IL1, IL3, IL6, Flt3

ligand, IL11

TGFβ, TNFα, INFγ, LIF

SELF RENEWAL COMMITTMENT PRECURSOR EXPANSION TERMINAL DIFFERENTIATION

Normal hematopoiesis

CFU-GM

CFU-M

CFU-G

CFU-MEG

Limphoid progenitor

Quiescent Staminal cell

Myeloblast

Monoblast

Megakaryoblast

Pro-erythroblast Erythroblast

Granulocyte

Monocyte

Platelets

Red cells

CFU-GEMM

BFU-E / CFU-E

Epo

SCF, TPO, IL3, GM-CSF

SCF, IL6 IL3, GM-CSF

Plasma cellB lymphocyte

Proliferating staminal cell

SCF, IL1, IL3, IL6, Flt3

ligand, IL11

TGFβ, TNFα, INFγ, LIF

SELF RENEWAL COMMITTMENT PRECURSOR EXPANSION TERMINAL DIFFERENTIATION

SOURCES OF HEMATOPOIETICSTEM CELLS FOR TRANSPLANT

Cord Blood 0.5%

Bone Marrow 1%

Mobilized Peripheral Blood 0.1-10%

THE CD34 ANTIGENFor more than a decade, CD34

molecule has been the best-known marker of HSC;Very high academic as commercial interest;

Berenson et al (1988) reported successful hematopoietic

reconstitution in baboons with selected CD34+ bone marrow

cells.

M.W. 110-120 kDaChromosome localization:1q32

E F M

TNF-alphaMIP-1 alphaTGF-beta

IL’sSCFLIFFlt3

LFA-1CD34

CD44

Thy1

L-selectinFlt3

c-kit

P-selectin

α5β1

α4β1

+ +- -

ICAM Collagen Fibronectin Laminin Proteoglycan VCAM-1Thrombospondin

Kit Rec Selectin RecFlt3 Rec

INTERACTIONS OF STEM CELLS WITH MICROENVRONMENT

HSC

Guitard…dello Sbarba et al.,Exp Hem 2010

HEMATOPOIETIC PROGENITOR CELLS

1950: MURINE MODELSCLINICAL INVESTIGATION

1970: COLONY ASSAYSTILL/ Mc CULLOGH

1975: BONE MARROW TRANSPLANTATION INHUMAN

1995: PHERIPHERAL BLOOD STEM CELLS

- Found throughout the body in all tissues in specific stromal niches

- Multipotent

- Partially committed with a key role in damaged tissues repair

The adult stem cell

- Derives from inner cell mass of blastocyst

- Pluripotent

- Give rise to endoderm, ectoderm, mesoderm tissues

The embryonic stem cell (ES)

Copyright Mike Jones

Diane S. Krause, Neil D. Theise, Michael I. Collector, Octavian Henegariu, Sonya Hwang, Rebekah Gardner, Sara Neutzel, and Saul J. Sharkis

Fluorescence microscopic imagesof (A) lung,(B) esophagus, (C) stomach, (D)colon, (E) bile duct cyst, (F) skin.

Donald Orlic, Jan Kajstura, Stefano Chimenti, Federica Limana, Igor Jakoniuk, FedericoQuaini, Bernardo Nadal-Ginard, David M. Bodine, Annarosa Leri, and Piero Anversa.

Mobilized bone marrow cells repair the infarcted heart, improving function and survival.

La cellula staminale emopoietica

PROPRIETA’ DELLA CELLULA STAMINALE

EMOPOIETICA:

• capacità di “self-renewal”

• capacità di dare origine a progenitori multipotenti

• capacità di dare origine a precursori “committed”

(differentiation plasticity)

• Capacità di engraftment

• Plasticità (developmental plasticity)

EVIDENZE SPERIMENTALI DELLA PLASTICITA’ DELLE CSE: UOMO

CUOREBELTRAMI (2001): I CARDIOMIOCITI POSSONO REPLICARE DOPO IMA

QUAINI (2002): MIGRAZIONE DI HSC DEL RICEVENTE NEL CUORE TRAPIANTATO

FEGATOALISON (2000) E THIESE (2000): STUDIO SU BIOPSIE EPATICHE DI DONNE TRAPIANTATE CON HSC DA UOMINI E BIOPSIE EPATICHE DI UOMINI TRAPIANTATI

CON FEGATI DI DONNE: CELLULE DI ORIGINE MIDOLLARE (CROMOSOMA Y) TROVATE NELL’1-40% DEI CASI.

RENEPOULSOM (2001): BIOPSIE RENALI DI DONNE CHE RICEVONO MIDOLLO OSSEO DA

UOMINI E DI UOMINI CHE RICEVONO RENI DA DONNE: RISCONTRO DI CELLULE DI ORIGINE MIDOLLARE NEL 34% DEI CASI.

GUPTA (2002): UOMINI CON IRA RICEVONO RENI DA DONNE: LE CELLULE DEL RICEVENTE SI TROVANO NEL RENE TRAPIANTATO.

- Derives from a genetic reprogramming of adult somatic cell

- Pluripotent artificial stem cell

- Different growth conditions lead to the differentiation into all possible types of specialized cells

The iPS: induced Pluripotent Stem cell

Skin fibroblasts (KeratinocyteBMSC, PBC)

Retroviral infection with Oct4, Sox2, c-Myc and Klf4 genes

iPS

Epithelium Cartilage Muscle CNS Adipose

(Reprogramming of human somatic cells to pluripotency with defined factors. Daley GQ et al., Nature, 2008)