Simulated Microgravity Potentiates Hematopoietic Differentiation of Human Pluripotent Stem Cells and Supports Formation of 3D Hematopoietic Cluster

Chiyuan Ma; Yue Xiong; Pei Han; Xueying Zhang; Yujing Cao; Baobei Wang; Huashan Zhao; Enkui Duan; Jian V Zhang; Xiaohua Lei

doi:10.3389/fcell.2021.797060

Simulated Microgravity Potentiates Hematopoietic Differentiation of Human Pluripotent Stem Cells and Supports Formation of 3D Hematopoietic Cluster

Front Cell Dev Biol. 2022 Jan 10:9:797060. doi: 10.3389/fcell.2021.797060. eCollection 2021.

Authors

Chiyuan Ma¹, Yue Xiong¹, Pei Han², Xueying Zhang¹, Yujing Cao³, Baobei Wang¹, Huashan Zhao¹, Enkui Duan³, Jian V Zhang^{1

4}, Xiaohua Lei^{1

4}

Affiliations

¹ Center for Energy Metabolism and Reproduction, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
² Technology and Engineering Center for Space Utilization, Chinese Academy of Sciences, Beijing, China.
³ State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
⁴ Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.

Abstract

Microgravity has been shown to induces many changes in proliferation, differentiation and growth behavior of stem cells. Little is known about the effect of microgravity on hematopoietic differentiation of pluripotent stem cells (PSCs). In this study, we used the random position machine (RPM) to investigate whether simulated microgravity (SMG) allows the induction of hematopoietic stem/progenitor cell (HSPC) derived from human embryonic stem cells (hESCs) in vitro. The results showed that SMG facilitates hESCs differentiate to HSPC with more efficient induction of CD34⁺CD31⁺ hemogenic endothelium progenitors (HEPs) on day 4 and CD34⁺CD43⁺ HSPC on day 7, and these cells shows an increased generation of functional hematopoietic cells in colony-forming unit assay when compared with normal gravity (NG) conditions. Additionally, we found that SMG significantly increased the total number of cells on day 4 and day 7 which formed more 3D cell clusters. Transcriptome analysis of cells identified thousands of differentially expressed genes (DEGs) between NG and SMG. DEGs down-regulated were enriched in the axonogenesis, positive regulation of cell adhesion, cell adhesion molecule and axon guidance, while SMG resulted in the up-regulation of genes were functionally associated with DNA replication, cell cycle, PI3K-Akt signaling pathway and tumorigenesis. Interestingly, some key gene terms were enriched in SMG, like hypoxia and ECM receptor interaction. Moreover, HSPC obtained from SMG culture conditions had a robust ability of proliferation in vitro. The proliferated cells also had the ability to form erythroid, granulocyte and monocyte/macrophage colonies, and can be induced to generate macrophages and megakaryocytes. In summary, our data has shown a potent impact of microgravity on hematopoietic differentiation of hPSCs for the first time and reveals an underlying mechanism for the effect of SMG on hematopoiesis development.

Keywords: 3D clusters; hematopoietic differentiation; human embryonic stem cells; microgravity; transcriptome analysis.