Previous Article in Journal
Reduced RhoGDI2 Expression Disrupts Centrosome Functions and Promotes Mitotic Errors
Previous Article in Special Issue
Integrating Macrophages into Human-Engineered Cardiac Tissue
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Cells
Volume 14
Issue 22
10.3390/cells14221835
cells-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Chemically Defined, Efficient Megakaryocyte Production from Human Pluripotent Stem Cells

by
Jae Eun Kim
1,
Yeonmi Lee
1,2,
Yonghee Kim
1,
Sae-Byeok Hwang
1,
Yoo Bin Choi
1,
Jongsuk Han
2,
Juyeol Jung
3,
Jae-woo Song
4,
Je-Gun Joung
2,
Jeong-Jae Ko
1 and
Eunju Kang
1,2,*
1
CHA R&D Institute, CHA Bundang Medical Center, Seongnam 13488, Republic of Korea
2
Department of Biomedical Science, College of Life Science, CHA University, Seongnam 13488, Republic of Korea
3
CHA R&D Institute, Artificial Intelligence Omics Research Center, Seongnam 13488, Republic of Korea
4
Department of Laboratory Medicine, Yonsei University, College of Medicine, Seodaemoon-gu, Seoul 03722, Republic of Korea
*
Author to whom correspondence should be addressed.
Cells 2025, 14(22), 1835; https://doi.org/10.3390/cells14221835 (registering DOI)
Submission received: 22 September 2025 / Revised: 24 October 2025 / Accepted: 17 November 2025 / Published: 20 November 2025
(This article belongs to the Special Issue Immune Cells from Pluripotent Stem Cells)
Review Reports Versions Notes

Abstract

Platelet shortage poses a significant barrier to research and transfusion therapies because native megakaryocytes (MKs) are scarce in blood. To overcome this limitation, pluripotent stem cell–derived MKs (PSC-MKs) offer a standardized, donor-independent platform for research and therapeutic development, including disease modeling and ex vivo platelet production. Here, we report a chemically defined, feeder-free protocol to generate MKs from human pluripotent stem cells (hPSCs). The protocol combines the small molecule MPL agonist Butyzamide, macrophage colony-stimulating factor (M-CSF), and three-dimensional (3D) suspension culture, achieving high efficiency and reproducibility. Butyzamide replaced recombinant thrombopoietin (TPO), yielding comparable CD41+/CD42b+ populations and enhanced polyploidization. M-CSF accelerated nuclear lobulation and induced 4N MKs, while 3D culture increased yield, cell size, and substrate detachment. Multiple independent assays confirmed mature MK hallmarks, multi-nuclei, demarcation membranes, granules, and elevated mitochondrial respiration. Single-cell RNA sequencing outlined a continuous trajectory from early progenitors to functionally specialized MK subsets. This platform enables reliable MK supply for mechanistic studies and in vitro platelet production, advancing both basic research and therapeutic development.
Keywords: pluripotent stem cells; megakaryocyte differentiation; Butyzamide; M-CSF; 3D suspension culture pluripotent stem cells; megakaryocyte differentiation; Butyzamide; M-CSF; 3D suspension culture

Share and Cite

MDPI and ACS Style

Kim, J.E.; Lee, Y.; Kim, Y.; Hwang, S.-B.; Choi, Y.B.; Han, J.; Jung, J.; Song, J.-w.; Joung, J.-G.; Ko, J.-J.; et al. Chemically Defined, Efficient Megakaryocyte Production from Human Pluripotent Stem Cells. Cells 2025, 14, 1835. https://doi.org/10.3390/cells14221835

AMA Style

Kim JE, Lee Y, Kim Y, Hwang S-B, Choi YB, Han J, Jung J, Song J-w, Joung J-G, Ko J-J, et al. Chemically Defined, Efficient Megakaryocyte Production from Human Pluripotent Stem Cells. Cells. 2025; 14(22):1835. https://doi.org/10.3390/cells14221835

Chicago/Turabian Style

Kim, Jae Eun, Yeonmi Lee, Yonghee Kim, Sae-Byeok Hwang, Yoo Bin Choi, Jongsuk Han, Juyeol Jung, Jae-woo Song, Je-Gun Joung, Jeong-Jae Ko, and et al. 2025. "Chemically Defined, Efficient Megakaryocyte Production from Human Pluripotent Stem Cells" Cells 14, no. 22: 1835. https://doi.org/10.3390/cells14221835

APA Style

Kim, J. E., Lee, Y., Kim, Y., Hwang, S.-B., Choi, Y. B., Han, J., Jung, J., Song, J.-w., Joung, J.-G., Ko, J.-J., & Kang, E. (2025). Chemically Defined, Efficient Megakaryocyte Production from Human Pluripotent Stem Cells. Cells, 14(22), 1835. https://doi.org/10.3390/cells14221835

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop