Previously undetected human hematopoietic cell populations with short-term repopulating activity selectively engraft NOD/SCID-β2 microglobulin–null mice
H. Glimm, … , C. von Kalle, C.J. Eaves
H. Glimm, … , C. von Kalle, C.J. Eaves
Published January 15, 2001
Citation Information: J Clin Invest. 2001;107(2):199-206. https://doi.org/10.1172/JCI11519.
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Article

Previously undetected human hematopoietic cell populations with short-term repopulating activity selectively engraft NOD/SCID-β2 microglobulin–null mice

Abstract

Increasing use of purified or cultured human hematopoietic cells as transplants has revealed an urgent need for better methods to predict the speed and durability of their engraftment potential. We now show that NOD/SCID-β2 microglobulin–null (NOD/SCID-β2m–/–) mice are sequentially engrafted by two distinct and previously unrecognized populations of transplantable human short-term repopulating hematopoietic cells (STRCs), neither of which efficiently engraft NOD/SCID mice. One is predominantly CD34+CD38+ and is myeloid-restricted; the other is predominantly CD34+CD38– and has broader lymphomyeloid differentiation potential. In contrast, the long-term repopulating human cells that generate lymphoid and myeloid progeny in NOD/SCID mice engraft and self-renew in NOD/SCID-β2m–/– mice equally efficiently. In short-term expansion cultures of adult bone marrow cells, myeloid-restricted STRCs were preferentially amplified (greater than tenfold) and, interestingly, both types of STRC were found to be selectively elevated in mobilized peripheral blood harvests. These results suggest an enhanced sensitivity of STRCs to natural killer cell–mediated rejection. They also provide new in vivo assays for different types of human STRC that may help to predict the engraftment potential of clinical transplants and facilitate future investigation of early stages of human hematopoietic stem cell differentiation.

Authors

H. Glimm, W. Eisterer, K. Lee, J. Cashman, T.L. Holyoake, F. Nicolini, L.D. Shultz, C. von Kalle, C.J. Eaves

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Figure 3

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The distribution of 6-week repopulating activity in NOD/SCID-β2m–/– mice...
The distribution of 6-week repopulating activity in NOD/SCID-β2m–/– mice between the G0/G1 and S/G2/M fractions of 5-day human CB cell-expansion cultures is indistinguishable from that measured for total cells or other progenitors. CD34+ CB cells were cultured for 5 days in serum-free medium supplemented with SF, FL, IL-3, IL-6, and G-CSF. G0/G1 and S/G2/M cells were then isolated after DNA staining with Hoechst 33342. Approximately half of the fractionated cells were transplanted in NOD/SCID-β2m–/– mice immediately after their isolation. The other half of each fraction was first cultured for an additional 16 hours before being transplanted. (a) The proportion of engrafted mice (and the average level of engraftment) for the two populations compared (G0/G1 and S/G2/M). (b) The corresponding data for total cells, CD34+ cells, CFC, and LTC-IC in the G0/G1 (open bars) and S/G2/M (filled bars) fractions from the same experiments. All values shown are the mean ± SEM of data pooled from three experiments.