Center for iPS Cell Research and Application, Kyoto University, 53 Shogoin-Kawahara, Sakyo, Kyoto, 606-8507, Japan.
Department of Micro Engineering, Kyoto University, Kyoto Daigaku-Katsura C3, Nishikyo-ku, Kyoto, 615-8540, Japan.
Biomed Microdevices. 2020 May 6;22(2):34. doi: 10.1007/s10544-020-00488-2.
A fundamental limitation in the derivation of hematopoietic stem and progenitor cells is the imprecise understanding of human developmental hematopoiesis. Herein we established a multilayer microfluidic Aorta-Gonad-Mesonephros (AGM)-on-a-chip to emulate developmental hematopoiesis from pluripotent stem cells. The device consists of two layers of microchannels separated by a semipermeable membrane, which allows the co-culture of human hemogenic endothelial (HE) cells and stromal cells in a physiological relevant spatial arrangement to replicate the structure of the AGM. HE cells derived from human induced pluripotent stem cells (hiPSCs) were cultured on a layer of mesenchymal stromal cells in the top channel while vascular endothelial cells were co-cultured on the bottom side of the membrane within the microfluidic device. We show that this AGM-on-a-chip efficiently derives endothelial-to-hematopoietic transition (EHT) from hiPSCs compared with regular suspension culture. The presence of mesenchymal stroma and endothelial cells renders functional HPCs in vitro. We propose that the AGM-on-a-chip could serve as a platform to dissect the cellular and molecular mechanisms of human developmental hematopoiesis.
造血干/祖细胞的一个根本限制是对人类发育性造血的理解不够精确。在此，我们建立了一个多层微流控主动脉-性腺-中肾（AGM）芯片，以从多能干细胞模拟发育性造血。该装置由两层微通道组成，中间由半透膜隔开，允许人类造血内皮（HE）细胞和基质细胞在生理相关的空间排列中进行共培养，以复制 AGM 的结构。HE 细胞来源于人诱导多能干细胞（hiPSC），在上层通道的一层间充质基质细胞上培养，而血管内皮细胞在微流控装置的膜的底部侧共培养。我们表明，与常规悬浮培养相比，这种 AGM 芯片能够有效地从 hiPSC 中诱导出内皮-造血转化（EHT）。间充质基质和内皮细胞的存在使体外具有功能性造血祖细胞（HPC）。我们提出，AGM 芯片可以作为一个平台，用于剖析人类发育性造血的细胞和分子机制。
