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A portable platform for stepwise hematopoiesis from human pluripotent stem cells within PET-reinforced collagen sponges

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Abstract

Various systems for differentiating hematopoietic cells from human pluripotent stem cells (PSCs) have been developed, although none have been fully optimized. In this report, we describe the development of a novel three-dimensional system for differentiating hematopoietic cells from PSCs using collagen sponges (CSs) reinforced with poly(ethylene terephthalate) fibers as a scaffold. PSCs seeded onto CSs were differentiated in a stepwise manner with appropriate cytokines under serum-free and feeder-free conditions. This process yielded several lineages of floating hematopoietic cells repeatedly for more than 1 month. On immunohistochemical staining, we detected CD34+ cells and CD45+ cells in the surface and cavities of the CS. Taking advantage of the portability of this system, we were able to culture multiple CSs together floating in medium, making it possible to harvest large numbers of hematopoietic cells repeatedly. Given these findings, we suggest that this novel three-dimensional culture system may be useful in the large-scale culture of PSC-derived hematopoietic cells.

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Acknowledgments

We thank Y. Sasaki and M. Yamane for their technical assistance, H. Watanabe for the administrative assistance and M. Osawa, M. Yanagimachi, T. Tanaka, M. Matsui, and S. Tajima for critical discussions. We are grateful to the Center for Anatomical, Pathological and Forensic Medical Researches, Graduate School of Medicine, Kyoto University for the Immunohistochemistry (IHC) analysis. Funding was provided by grants from the Ministry of Health, Labour and Welfare to T.N. and M.K.S., grants from the Leading Project of MEXT to T.N., a grant from the Funding Program for World-Leading Innovative Research and Development on Science and Technology (FIRST Program) of the Japan Society for the Promotion of Science (JSPS) to T.N., JSPS KAKENHI Grant No. JP25860856 to A.N., Japan Science and Technology Agency CREST to T.N., the Program for Intractable Diseases Research utilizing disease-specific iPS cells from Japan Agency for Medical Research and Development (AMED) to T.N., the grant for Core Center for iPS Cell Research of Research Center Network for Realization of Regenerative Medicine from AMED to T.N. and M.K.S, and grants from the JSPS to T.N. and M.K.S. All of the funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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    Authors and Affiliations

    1. Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto, 606-8507, Japan

      Yoshinori Sugimine, Akira Niwa, Hiroyuki Matsubara, Kazuki Kobayashi, Tatsutoshi Nakahata & Megumu K. Saito

    2. Department of Pediatrics, Kyoto University Graduate School of Medicine, Kyoto, 6068507, Japan

      Yoshinori Sugimine & Toshio Heike

    3. Department of Biomaterials, Field of Tissue Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, 6068507, Japan

      Yasuhiko Tabata

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    1. Yoshinori Sugimine
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    4. Kazuki Kobayashi
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    5. Yasuhiko Tabata
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    7. Tatsutoshi Nakahata
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    Correspondence to Megumu K. Saito.

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    Y. Sugimine and A. Niwa contributed equally to this work.

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    Sugimine, Y., Niwa, A., Matsubara, H. et al. A portable platform for stepwise hematopoiesis from human pluripotent stem cells within PET-reinforced collagen sponges. Int J Hematol 104, 647–660 (2016). https://doi.org/10.1007/s12185-016-2088-x

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