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Stromal cells from murine embryonic aorta–gonad–mesonephros region, liver and gut mesentery expand human umbilical cord blood-derived CAFCweek6 in extended long-term cultures

Leukemia volume 16, pages 1782–1790 (2002)Cite this article

Abstract

The first definitive long-term repopulating hematopoietic stem cells (HSCs) emerge from and undergo rapid expansion in the embryonic aorta–gonad–mesonephros (AGM) region. To investigate the presumptive unique characteristics of the embryonic hematopoietic microenvironment and its surrounding tissues, we have generated stromal clones from subdissected day 10 and day 11 AGMs, embryonic livers (ELs) and gut mesentery. We here examine the ability of 19 of these clones to sustain extended long-term cultures (LTCs) of human CD34+ umbilical cord blood (UCB) cells in vitro. The presence of in vitro repopulating cells was assessed by sustained production of progenitor cells (extended LTC-CFC) and cobblestone area-forming cells (CAFC). The embryonic stromal clones differed greatly in their support for human HSCs. Out of eight clones tested in the absence of exogenous cytokines, only one (EL-derived) clone was able to provide maintenance of HSCs. Addition of either Tpo or Flt3-L + Tpo improved the long-term support of about 50% of the tested clones. Cultures on four out of 19 clones, ie the EL-derived clone mentioned, two urogenital-ridge (UG)-derived clones and one gastrointestinal (GI)-derived clone, allowed a continuous expansion of primitive CAFC and CFU-GM with over several hundred-fold more CAFCweek6 produced in the 12th week of culture. This expansion was considerably higher than that found with the FBMD-1 cell line, which is appreciated by many investigators for its support of human HSCs, under comparable conditions. This stromal cell panel derived from the embryonic regions may be a powerful tool in dissecting the factors mediating stromal support for maintenance and expansion of HSCs.

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Acknowledgements

We would like to acknowledge the nursing staff and midwives of the St Fransiscus Gasthuis for collection of UCB samples, and PB van Hennik, WJC Rombouts and I Blokland for technical assistance. Supported by research grants of the NIH R01 DK51077 (ED) and AMGEN, Thousand Oaks, CA (RO, ED).

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Authors and Affiliations

  1. Institute of Hematology, Erasmus Medical Center Rotterdam, The Netherlands

    N Kusadasi, WJLM Koevoet & RE Ploemacher

  2. Cell Biology and Genetics, Erasmus Medical Center Rotterdam, The Netherlands

    RAJ Oostendorp & EA Dzierzak

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Kusadasi, N., Oostendorp, R., Koevoet, W. et al. Stromal cells from murine embryonic aorta–gonad–mesonephros region, liver and gut mesentery expand human umbilical cord blood-derived CAFCweek6 in extended long-term cultures. Leukemia 16, 1782–1790 (2002). https://doi.org/10.1038/sj.leu.2402615

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