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Table I

HSC engraftment capacity as a function of cell cycle status




3–4 wk

9–12 wk

>18 wk
Phase
Cell #
Frequencyreconstitution(#/total)
Average chimerism(range)engraftment type(#/reconstituted)
Frequencyreconstitution(#/total)
Average chimerism(range)engraftment type(#/reconstituted)
Frequencyreconstitution(#/total)
Average chimerism(range)engraftment type(#/reconstituted)
LT-HSC        
G0 10 57% (8/14) 1.1% (0.1–3.9%)
 M + L (8/8) 57% (8/14) 4.7% (0.3–15.2%)
 M + L (8/8) 54% (7/13) 7.5% (0.6–23.7%)
 M + L (7/7) 
 50 100% (5/5) 1.5% (0.1–5.8%)
 M + L (5/5) 100% (5/5) 5.7% (0.5–14.3%)
 M + L (5/5) 100% (5/5) 11.8% (0.3–35.3%)
 M + L (4/5), L only (1/5) 
G1 10 8% (1/12) 0.2%
 M only (1/1) 0% (0/11) n/a 0% (0/11) n/a 
 50 50% (3/6) 4.2% (0.1–12.5%)
 M only (3/3) 0% (0/5) n/a 0% (0/5) n/a 
S-G2/M 10 0% (0/9) n/a 0% (0/9) n/a 0% (0/9) n/a 
 50 50% (3/6) 0.4% (0.2–0.7%)
 M + L (3/3) 33% (2/6) 0.35% (0.3–0.4%)
 M + L (1/2), L only (1/2) 33% (2/6) 0.15% (0.1–0.2%)
 L only (2/2) 
KTLS HSC        
G0 10 50% (6/12) 0.7% (0.1–1.2%)
 M + L (5/6), M only (1/6) 42% (5/12) 3% (0.2–10.6%)
 M + L (5/5) 42% (5/12) 3.2% (0.3–8.3%)
 M + L (4/5), L only (1/5) 
 50 100% (4/4) 0.7% (0.1–1.2%)
 M + L (4/4) 100% (4/4) 2.9% (0.4–8.2%)
 M + L (4/4) 100% (4/4) 3.0% (0.2–6.2%)
 M + L (4/4) 
G1 10 0% (0/10) n/a 0% (0/10) n/a 0% (0/10) n/a 
 50 25% (1/4) 0.3%
 M (1/1) 0% (0/4) n/a 0% (0/4) n/a 
S-G2/M 10 0% (0/10) n/a 0% (0/10) n/a 0% (0/10) n/a 

 
50
 
20% (1/5)
 
0.6%
 M + L (1/1)
 
20% (1/5)
 
0.3%
 L only (1/1)
 
0% (0/5)
 
n/a
 



3–4 wk

9–12 wk

>18 wk
Phase
Cell #
Frequencyreconstitution(#/total)
Average chimerism(range)engraftment type(#/reconstituted)
Frequencyreconstitution(#/total)
Average chimerism(range)engraftment type(#/reconstituted)
Frequencyreconstitution(#/total)
Average chimerism(range)engraftment type(#/reconstituted)
LT-HSC        
G0 10 57% (8/14) 1.1% (0.1–3.9%)
 M + L (8/8) 57% (8/14) 4.7% (0.3–15.2%)
 M + L (8/8) 54% (7/13) 7.5% (0.6–23.7%)
 M + L (7/7) 
 50 100% (5/5) 1.5% (0.1–5.8%)
 M + L (5/5) 100% (5/5) 5.7% (0.5–14.3%)
 M + L (5/5) 100% (5/5) 11.8% (0.3–35.3%)
 M + L (4/5), L only (1/5) 
G1 10 8% (1/12) 0.2%
 M only (1/1) 0% (0/11) n/a 0% (0/11) n/a 
 50 50% (3/6) 4.2% (0.1–12.5%)
 M only (3/3) 0% (0/5) n/a 0% (0/5) n/a 
S-G2/M 10 0% (0/9) n/a 0% (0/9) n/a 0% (0/9) n/a 
 50 50% (3/6) 0.4% (0.2–0.7%)
 M + L (3/3) 33% (2/6) 0.35% (0.3–0.4%)
 M + L (1/2), L only (1/2) 33% (2/6) 0.15% (0.1–0.2%)
 L only (2/2) 
KTLS HSC        
G0 10 50% (6/12) 0.7% (0.1–1.2%)
 M + L (5/6), M only (1/6) 42% (5/12) 3% (0.2–10.6%)
 M + L (5/5) 42% (5/12) 3.2% (0.3–8.3%)
 M + L (4/5), L only (1/5) 
 50 100% (4/4) 0.7% (0.1–1.2%)
 M + L (4/4) 100% (4/4) 2.9% (0.4–8.2%)
 M + L (4/4) 100% (4/4) 3.0% (0.2–6.2%)
 M + L (4/4) 
G1 10 0% (0/10) n/a 0% (0/10) n/a 0% (0/10) n/a 
 50 25% (1/4) 0.3%
 M (1/1) 0% (0/4) n/a 0% (0/4) n/a 
S-G2/M 10 0% (0/10) n/a 0% (0/10) n/a 0% (0/10) n/a 

 
50
 
20% (1/5)
 
0.6%
 M + L (1/1)
 
20% (1/5)
 
0.3%
 L only (1/1)
 
0% (0/5)
 
n/a
 

LT-HSC and KTLS HSC were first double sorted from C57BL/6-Ly5.1 donor mice, stained with H/PY, resorted for cell cycle G0 (H2N/PY), G1 (H2N/PY+), and S-G2/M (H>2N-4N/PY+) subpopulations and tested for their engraftment capacities by competitive reconstitution assays. 10 or 50 cells were transplanted into lethally irradiated congenic C57BL/6-Ly5.2 recipient mice, together with 3 × 105 Ly5.2+ helper BM cells. Recipient mice were considered to be reconstituted by Ly5.1+ donor cells if the frequencies of Ly5.1+ donor-derived myeloid (M, Gr-1+, or Mac-1+) or lymphoid (L, B220+, or CD3+) cells detected in the peripheral blood at the indicated times after transplantation were greater than background levels (0.1% as determined by parallel analysis of untransplanted control C57BL/6-Ly5.2 mice). M+L indicates multilineage reconstitution. n/a, not applicable.

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