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Stroma-Contact Prevents Loss of Hematopoietic Stem Cell Quality
During Ex Vivo Expansion of CD34+ Mobilized Peripheral
Blood Stem Cells
Dimitri A. Breems,
Ellen A.W. Blokland,
Karen E. Siebel,
Angelique
E.M. Mayen,
Lilian J.A. Engels, and
Rob E. Ploemacher
From the Institute of Hematology, Erasmus University Rotterdam,
Rotterdam, The Netherlands.
Stroma-supported long-term cultures (LTC) allow estimation of stem
cell quality by simultaneous enumeration of hematopoietic stem cell
(HSC) frequencies in a graft using the cobblestone area forming cell
(CAFC) assay, and the ability of the graft to generate progenitors in
flask LTC (LTC-CFC). We have recently observed that the number and
quality of mobilized peripheral blood stem cells (PBSC) was low in
patients having received multiple rounds of chemotherapy. Moreover,
grafts with low numbers of HSC and poor HSC quality had a high
probability to cause graft failure upon their autologous infusion.
Because ex vivo culture of stem cells has been suggested to present an
attractive tool to improve hematological recovery or reduce graft size,
we have studied the possibility that such propagation may affect stem
cell quality. In order to do so, we have assessed the recovery of
different stem cell subsets in CD34+ PBSC after a 7-day
serum-free liquid culture using CAFC and LTC-CFC assays. A numerical
expansion of stem cell subsets was observed in the presence of
interleukin-3 (IL-3), stem cell factor, and IL-6, while stroma-contact,
stromal soluble factors, or combined addition of FLT3-ligand and
thrombopoietin improved this parameter. In contrast, ex vivo culture
severely reduced the ability of the graft to produce progenitors in LTC
while stromal soluble factors partly abrogated this quality loss. The
best conservation of graft quality was observed when the PBSC were
cultured in stroma-contact. These data suggest that ex vivo propagation
of PBSC may allow numerical expansion of various stem cell subsets,
however, at the expense of their quality. In addition, cytokine-driven
PBSC cultures require stroma for optimal maintenance of graft quality.
Blood, Vol. 91 No. 1 (January 1), 1998:
pp. 111-117
© 1998 by The American Society of Hematology.
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