Self-organized yolk sac-like organoids allow for scalable generation of multipotent hematopoietic progenitor cells from induced pluripotent stem cells

Naritaka Tamaoki; Stefan Siebert; Takuya Maeda; Ngoc-Han Ha; Meghan L Good; Yin Huang; Suman K Vodnala; Juan J Haro-Mora; Naoya Uchida; John F Tisdale; Colin L Sweeney; Uimook Choi; Julie Brault; Sherry Koontz; Harry L Malech; Yasuhiro Yamazaki; Risa Isonaka; David S Goldstein; Masaki Kimura; Takanori Takebe; Jizhong Zou; David F Stroncek; Pamela G Robey; Michael J Kruhlak; Nicholas P Restifo; Raul Vizcardo

doi:10.1016/j.crmeth.2023.100460

Self-organized yolk sac-like organoids allow for scalable generation of multipotent hematopoietic progenitor cells from induced pluripotent stem cells

Cell Rep Methods. 2023 Apr 24;3(4):100460. doi: 10.1016/j.crmeth.2023.100460.

Authors

Naritaka Tamaoki^{1

2}, Stefan Siebert³, Takuya Maeda^{1

2}, Ngoc-Han Ha⁴, Meghan L Good^{1

2}, Yin Huang^{1

2}, Suman K Vodnala^{1

2}, Juan J Haro-Mora⁵, Naoya Uchida⁵, John F Tisdale⁵, Colin L Sweeney⁶, Uimook Choi⁶, Julie Brault⁶, Sherry Koontz⁶, Harry L Malech⁶, Yasuhiro Yamazaki⁷, Risa Isonaka⁸, David S Goldstein⁸, Masaki Kimura⁹, Takanori Takebe^{9

10}, Jizhong Zou¹¹, David F Stroncek¹², Pamela G Robey¹³, Michael J Kruhlak⁴, Nicholas P Restifo^{1

2}, Raul Vizcardo^{1

2}

Affiliations

¹ Surgery Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD 20892, USA.
² Center of Cell-based Therapy, National Cancer Institute, NIH, Bethesda, MD 20892, USA.
³ Department of Molecular and Cellular Biology, University of California, Davis, Davis, CA 95616, USA.
⁴ Laboratory of Cancer Biology and Genetics, National Cancer Institute, NIH, Bethesda, MD 20892, USA.
⁵ Cellular and Molecular Therapeutics Branch, National Heart, Lung, and Blood Institute/National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, MD 20892, USA.
⁶ Genetic Immunotherapy Section, Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA.
⁷ Immune Deficiency Genetics Section, Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA.
⁸ Autonomic Medicine Section, National Institute of Neurological Disorders and Stroke, NIH, Bethesda, MD 20892, USA.
⁹ Division of Gastroenterology, Hepatology & Nutrition, Developmental Biology, Center for Stem Cell and Organoid Medicine (CuSTOM), Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229-3039, USA.
¹⁰ Premium Research Institute for Human Metaverse Medicine (WPI-PRIMe), and Division of Stem Cell and Organoid Medicine, Osaka University, Suita, Osaka 565-0871, Japan.
¹¹ iPSC Core, National Heart, Lung, and Blood Institute, NIH, Bethesda, MD 20892, USA.
¹² Cell Processing Section, Department of Transfusion Medicine, Clinical Center, NIH, Bethesda, MD 20892, USA.
¹³ Skeletal Biology Section, National Institute of Dental and Craniofacial Research, NIH, Bethesda, MD 20892, USA.

Abstract

Although the differentiation of human induced pluripotent stem cells (hiPSCs) into various types of blood cells has been well established, approaches for clinical-scale production of multipotent hematopoietic progenitor cells (HPCs) remain challenging. We found that hiPSCs cocultured with stromal cells as spheroids (hematopoietic spheroids [Hp-spheroids]) can grow in a stirred bioreactor and develop into yolk sac-like organoids without the addition of exogenous factors. Hp-spheroid-induced organoids recapitulated a yolk sac-characteristic cellular complement and structures as well as the functional ability to generate HPCs with lympho-myeloid potential. Moreover, sequential hemato-vascular ontogenesis could also be observed during organoid formation. We demonstrated that organoid-induced HPCs can be differentiated into erythroid cells, macrophages, and T lymphocytes with current maturation protocols. Notably, the Hp-spheroid system can be performed in an autologous and xeno-free manner, thereby improving the feasibility of bulk production of hiPSC-derived HPCs in clinical, therapeutic contexts.

Keywords: HPC; bulk production; hematopoiesis; human iPSC; organoid; stromal cell; xeno-free condition; yolk sac.

Publication types

Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural
Research Support, Non-U.S. Gov't

MeSH terms

Activities of Daily Living
Hematopoietic Stem Cells
Humans
Induced Pluripotent Stem Cells*
Organoids
Yolk Sac

Abstract

Publication types

MeSH terms

Grants and funding