A portable platform for stepwise hematopoiesis from human pluripotent stem cells within PET-reinforced collagen sponges.
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.
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto, 606-8507, Japan.|Department of Pediatrics, Kyoto University Graduate School of Medicine, Kyoto, 6068507, Japan.
Yoshinori
Y
Sugimine
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto, 606-8507, Japan.
Akira
A
Niwa
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto, 606-8507, Japan.
Hiroyuki
H
Matsubara
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto, 606-8507, Japan.
Kazuki
K
Kobayashi
Department of Biomaterials, Field of Tissue Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, 6068507, Japan.
Yasuhiko
Y
Tabata
Department of Pediatrics, Kyoto University Graduate School of Medicine, Kyoto, 6068507, Japan.
Toshio
T
Heike
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto, 606-8507, Japan.
Tatsutoshi
T
Nakahata
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto, 606-8507, Japan. msaito@cira.kyoto-u.ac.jp.
Megumu K
http://orcid.org/0000-0001-8813-3614
MK
Saito
D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry*; D004867:Equipment Design; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D000066449:Human Embryonic Stem Cells / Q000166:cytology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000166:cytology*; D011093:Polyethylene Terephthalates / Q000737:chemistry*; D054457:Tissue Scaffolds / Q000737:chemistry*
D011093:Polyethylene Terephthalates; D003094:Collagen
Collagen sponge; Hematopoietic differentiation; Hematopoietic progenitor cells; Human embryonic stem cells; Human induced pluripotent stem cells