|
|
Blood, Vol. 108, Issue 4, 1230-1233, August 15, 2006
The p67 laminin receptor identifies human erythroid progenitor and precursor cells and is functionally important for their bone marrow lodgment
Blood Bonig et al.
108: 1230
Supplemental materials for: Bonig et al
Files in this Data Supplement:
- Figure S1. p67— expression during erythroid maturation (PDF, 59.9 KB) -
TOP: p67 is expressed on the majority of proerythroblasts, which were picked from day-7 BFU-E. Colonies were identified by their typical morphology, and identity as proerythroblasts was confirmed by CD45—low GlycophorinA+ staining on FACS, by cell morphology in Wright-Giemsa stained cytospins, and by negativity for Benzidine staining. Most of these cells expressed p67.
BOTTOM: p67 is not well expressed on erythroblasts picked from day-14 BFU-E. Day-14 BFU-E were hemoglobinized and showed the characteristic colony morphology. The cells were CD45-negative and GlycophorinA+ on FACS, showed the morphology of late erythroblasts, and stained positive for Benzidine.
Microscope Olympus BH2, 40× original magnification, Nikon CoolPix 995 camera, unadjusted images.
- Figure S2 (PDF, 13.9 KB) -
A: CFU-C frequency in sorted CD34+p67— and CD34+p67+ cell samples: Results for the individual experiments are shown. Three experiments were done with cryopreserved BM, one experiment each with frozen and fresh MPB. In all experiments, BFU-E were significantly enriched in the p67+ cell fraction. * only one plate was available for analysis (Colony number/1000 plated cells; mean ± SEM).
B: Blockade of MPB-derived BFU-E homing to marrow by anti-p67Ab: Marrow homing of MPB CFU-C was inhibited by anti-p67Ab incubation, predominantly at the expense of BFU-E. BM homing of total CFU-C, BFU E, CFU-mixed and CFU GM was reduced by 61.2 ± 3.0%, 76.8 ± 3.0%, 79.2 ± 5.9% and 45.0 ± 4.0%, respectively (p < 0.005; mean + SEM of % of the total injected CFU-C homed to marrow 20 h after transplantation; n=1 experiment, 5 + 5 recipients). Our data thus suggest that, in contrast to BM, a fraction of MPB CFU-GM may express functional levels of p67.
- Figure S3. p67+ BM cells are enriched for BFU-E (PDF, 36.6 KB) -
Aliquots of sorted p67— or p67+ BM cells or unsorted BM cells were plated in plasma clot assays. Plasma clots were harvested after 11 days in culture and Benzidine-stained to enumerate BFU-E. Compared to unsorted controls, BFU-E were significantly depleted in the p67— fraction and significantly enriched in the p67+ fraction. Note that the starting population were total BM nucleated cells, rather than CD34+ cells (unsorted: 10000 cells/clot, p67— and p67+: 5000 cells/clot; 2 of 5 replicates are shown). Leica MZ6 microscope, 0.63× magnification, Nikon CoolPix 995 camera, 2× Zoom.
|
|
