Adapted from Gazit R Weissman IL and Rossi DJ 2008 HSC MPP Mouse Lineage low Sca1 cKit flk2 CD34 CD48 CD150 Human Lineage low CD90 CD45RA CD34 ID: 387305
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Slide1
Hematopoietic Stem Cells (HSC)
Adapted from {Gazit R, Weissman IL and Rossi DJ 2008}
HSC
MPP
Mouse
Lineage
-/low
Sca1
+cKit+flk2-CD34-CD48-CD150+
HumanLineage-/lowCD90+CD45RA-CD34+CD38-CD49f+
Functional definition
Repopulate Irradiated RecipientsGenerate all Blood cell types (Peripheral Blood myeloid and lymphoid)Long-term (>4month)
Multipotent
Self-
RenewSlide2
Stem and Progenitors Cells
form a Population within the Hematopoietic system
HSPC
DC
Mo
MF
GN
preT
B
NKT
NK
T
Tgd
proB
Unguided
transcriptome
PCA found clear clustering of cell typesSlide3
HSC-specific
Transcription regulators include known and novel genes – that cluster togetherSlide4
HSC specific Transcription Factors
(and other genes)
Lymphoid- and Myeloid- Progenitors
Mo.
Mac
Ost
.
DC
MPP2
HSC
B cells
GN
NK
T Cells
Multipotent
Eryth
.
Oligo
-potent
Lineage-committed and effector cells
MPP1
Glis2
Hoxa9
Hoxa5
Rbpms
Prdm16
Zfp467
Mycn
Pbx1
Irf6
Ndn
Hoxb5
Rarb
Zfp37
Hlf
Meis1
Msi2
Zfp532
Nap1l3
Zfp521
Zfp612Slide5
One Transcription Factor,
Hlf
,
induce HSC-markers and sustain
multilineages
Percent Lin- CD150+
time (weeks)
0
1
2
3
HLF
ZsGreen
HSC
MPPflk2-
MPPflk2+
GMP
CMP
MEP
b
c
HSC
+HLF
HSC
+dsRed
control
mac.
meg.
mac.
prog.
gran.
mac.
+ZsGreen
+HLF
a
CD11b
CD150
MPPflk2-
HSC
MPPflk2+
GMP
CMP
MEP
MPPflk2-
HSC
MPPflk2+
GMP
CMP
MEP
0
10
2
10
3
10
4
10
5
0
10
2
10
3
10
4
10
5
0
10
2
10
3
10
4
10
5
0
10
2
10
3
10
4
10
5
1.1±0.4
2.3±0.4
0.8±0.4
13.2±3.0
6.0±2.3
22.1±10.3
55.4±2.2
60.2±7.2
56.2±5.3
12.4±12.1
60.8±18.4
63.6±5.5
FACS at 2weeksSlide6
Eight Transcription Factors can reprogram progenitors into long-term repopulating cells
3.32
30.1
66.4
CD45.2
CD45.1
0.83
0.21
58.5
40.4
CD4+CD8+GR1
Mac1
CD45.1
ZsGreen
(Control)
CD45.2
With 8 TFs
10.9
48.9
9.51
30.6
Eto
Hoxb5
Hlf
Irf6
Msi2
Nap1l3
Rbpms
Zfp467Slide7
Knockin
mouse with Fgd5 mCherry
label HSC
FSC
mCherry
0
10
2
10
3
10
4
10
5
0
10
2
10
3
10
4
10
5
99%
0
10
2
10
3
10
4
10
5
0
10
2
10
3
10
4
10
5
98%
0
10
2
10
3
10
4
10
5
0
10
2
10
3
10
4
10
5
94%
FSC
cKit
CD48
0
50K
100K
150K
200K
250K
0
10
2
10
3
10
4
10
5
0.035%
99.9%
Pre-gated on Live,
doublet-discriminated, BM cells
Gating on
mCherry
+ cells identify HSCs
Lineage
Sca1
CD150
mCherry
+ cells are functional HSCs, and all of the HSCs.
Weeks Post-Transplant
0.1
1
10
100
4
8
12
16
20
24
Donor
chimerism
(%)
FSC
mCherry
0
50K
100K
150K
200K
250K
0
10
2
10
3
10
4
10
5
99.9%
0.03%
Fgd5
+/
mCherry
100
mCherry
+
100,000
mCherry
-