iPSC-derived mesenchymal stromal cells are less supportive than primary MSCs for co-culture of hematopoietic progenitor cells
- Theresa Vasko†1, 2,
- Joana Frobel†1, 2,
- Richard Lubberich1, 2,
- Tamme W. Goecke3 and
- Wolfgang Wagner1, 2Email author
© Vasko et al. 2016
Received: 2 March 2016
Accepted: 13 April 2016
Published: 21 April 2016
In vitro culture of hematopoietic stem and progenitor cells (HPCs) is supported by a suitable cellular microenvironment, such as mesenchymal stromal cells (MSCs)—but MSCs are heterogeneous and poorly defined. In this study, we analyzed whether MSCs derived from induced pluripotent stem cells (iPS-MSCs) provide a suitable cellular feeder layer too. iPS-MSCs clearly supported proliferation of HPCs, maintenance of a primitive immunophenotype (CD34+, CD133+, CD38-), and colony-forming unit (CFU) potential of CD34+ HPCs. However, particularly long-term culture-initiating cell (LTC-IC) frequency was lower with iPS-MSCs as compared to primary MSCs. Relevant genes for cell-cell interaction were overall expressed at similar level in MSCs and iPS-MSCs, whereas VCAM1 was less expressed in the latter. In conclusion, our iPS-MSCs support in vitro culture of HPCs; however, under the current differentiation and culture conditions, they are less suitable than primary MSCs from bone marrow.
We assessed the CFU frequency in freshly isolated HPCs or upon culture-expansion for 7 days: without stromal support, there was no expansion of CFUs, whereas CFU frequency was significantly increased under co-culture conditions with MSCs or iPS-MSCs (Fig. 1d). CFU frequency was not significantly affected if HPCs were co-cultured either with MSCs or iPS-MSCs, and there was no bias towards specific types of colonies (Fig. 1d). However, if HPCs were cultured for 5 weeks in a long-term culture-initiating cell (LTC-IC) assay , different hematopoiesis supporting capacities of MSCs and iPS-MSCs became evident: long-term culture of HPCs gave rise to a significantly higher number of colonies on MSCs compared to iPS-MSCs (Fig. 1e).
Taken together, our iPS-MSCs provide a less hematopoiesis supportive microenvironment than primary MSCs, particularly after long-term co-culture. This tendency was not observed in a recent study by Moslem et al. , but these authors did not test for maintenance of LTC-ICs. It is conceivable that optimized differentiation procedures of iPSCs towards cellular components of the hematopoietic stem cell niche as well as 3D-culture systems will further enhance stromal support to ultimately facilitate in vitro expansion of HPCs.
This work was supported by the Else Kröner-Fresenius Stiftung (2014_A193), by the German Research Foundation (WA/1706/2-1), and by the Interdisciplinary Center for Clinical Research (IZKF) in the Faculty of Medicine at the RWTH Aachen University (O1-1).
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