Modeling Early Post-Implantation Development

Modeling Neurodevelopment

Mechanobiology of Human Pluripotent Stem Cells

Cell Mechanics and Mechanotransduction

 

Modeling Early Post-Implantation Development

Early post-implantation human development remains mysterious and very difficult to study. Recent advances in the development of human embryo models from human stem cells, including both human embryonic and extraembryonic stem cells, have sparked great interest in developing and applying such models to advance knowledge of human development and regenerative and reproductive medicine. In this research, we have leveraged the developmental potential and self-organizing property of human stem cells in conjunction with biomimetic culture systems to develop different embryo models to study the early post-implantation human development. Specifically, we have successfully developed the first human embryo model that recapitulates successive embryogenic events during the early post-implantation human development, including amniotic cavity formation, amnion-epiblast patterning, specification of primordial germ cells, and the onset of gastrulation. The long-term goal of our research is to integrate bioengineering innovations with stem cell biology and developmental biology to establish faithful and useful experimental models of the early post-implantation human development. These models are promising in revealing new knowledge of some previously inaccessible phases of human development, including the implantation, placentation, gastrulation, and early organogenesis.


During the first three weeks of human development, a few key embryogenic events occur, including blastocyst formation, implantation and gastrulation. Gastrulation is the fundamental organizational event that generates the basic body plan and provides the building blocks for all the tissues in the human embryo.

Selected Publications:

  1. Jun Wu and Jianping Fu. Towards developing human organs via embryo models and chimeras. Cell, vol. 187, pp. 3194-3219, 2024. [PDF]
  2. Sajedeh Nasr Esfahani, Yi Zheng, Auriana Arabpour, Agnes M. Resto Irizarry, Norio Kobayashi, Xufeng Xue, Yue Shao, Cheng Zhao, Nicole L. Agranonik, Megan Sparrow, Timothy J. Hunt, Jared Faith, Mary Jasmine Lara, Qiu Ya Wu, Sherman Silber, Sophie Petropoulos, Ran Yang, Kenneth R. Chien, Amander T. Clark, and Jianping Fu. Derivation of human primordial germ cell-like cells in an embryonic-like culture. Nature Communications, vol. 15, 167, 2024. [PDF | Supplemental Materials]
  3. Nicolas C. Rivron, Alfonso Martinez-Arias, Karen Sermon, Christine Mummery, Hans Schöler, James Wells, Jenny Nichols, Anna-Katerina Hadjantonakis, Madeline A. Lancaster, Jianping Fu, Janet Rossant, and Kazuto Kato. Changing the public perception of human embryology. Nature Cell Biology, vol. 25, pp. 1717-1719, 2023. [PDF]
  4. Janet Rossant and Jianping Fu. Why researchers should use human embryo models with caution. Nature, vol. 622, pp. 22-24, 2023. [PDF]
  5. Zongyong Ai, Baohua Niu, Yu Yin, Lifeng Xiang, Gaohui Shi, Kui Duan, Sile Wang, Yingjie Hu, Chi Zhang, Chengting Zhang, Lujuan Rong, Ruize Kong, Tingwei Chen, Yixin Guo, Wanlu Liu, Nan Li, Shumei Zhao, Xiaoqing Zhu, Xuancheng Mai, Yonggang Li, Ze Wu, Yi Zheng, Jianping Fu, Weizhi Ji, and Tianqing Li. Dissecting peri-implantation development using cultured human embryos and embryo-like assembloids. Cell Research, vol. 33, pp. 661-678, 2023. [PDF | Supplemental Materials]
  6. Yi Zheng, Robin Zhexuan Yan, Shiyu Sun, Mutsumi Kobayashi, Lifeng Xiang, Ran Yang, Alexander Goedel, Yu Kang, Xuefeng Xue, Sajedeh Nasr Esfahani, Yue Liu, Agnes M. Resto Irizarry, Weisheng Wu, Yunxiu Li, Weizhi Ji, Yuyu Niu, Kenneth R. Chien, Tianqing Li, Toshihiro Shioda, and Jianping Fu. Single-cell analysis of embryoids reveals lineage diversification roadmaps of early human development. Cell Stem Cell, vol. 29, pp. 1402-1419, 2022. [PDF | Supplemental Materials]
  7. Yue Shao, and Jianping Fu. Engineering multiscale structural orders for high-fidelity embryoids and organoids. Cell Stem Cell, vol. 29, pp. 722-743, 2022. [PDF]
  8. Ran Yang, Alexander Goedel, Yu Kang, Chengyang Si, Chu Chu, Yi Zheng, Zhenzhen Chen, Peter J. Gruber, Yao Xiao, Chikai Zhou, Chuen-Yan Leung, Yongchang Chen, Jianping Fu, Weizhi Ji, Fredrik Lanner, Yuyu Niu, and Kenneth Chien. Amnion signals are essential for mesoderm formation in primates. Nature Communications, vol. 12, 5126, 2021. [PDF]
  9. Sicong Wang, Chien-Wei Lin, Chari L. Cortez, Amber E. Carleton, Craig Johnson, Linnea E. Taniguchi, Ryan F. Townshend, Venkatesha Basrur, Alexey I. Nesvizhskii, Amy W. Hudson, Blake R. Hill, Peng Zou, Jianping Fu, Deborah L. Gumucio, Mara C. Duncan, and Kenichiro Taniguchi. Spatially resolved cell polarity proteomics of a human epiblast model. Science Advances, vol. 7, eabd8407, 2021. [PDF]
  10. Jianping Fu, Aryeh Warmflash, and Lutolf P. Matthias. Stem-cell-based embryo models for fundamental research and translation. Nature Materials, vol. 20, pp. 132-144, 2021. [PDF]
  11. Jonathon M. Muncie, Nadia M.E. Ayad, Johnathon N. Lakins, Xufeng Xue, Jianping Fu, and Valerie M. Weaver. Mechanical tension promotes formation of gastrulation-like nodes and patterns mesoderm specification in human embryonic stem cells. Developmental Cell, vol. 55, pp. 679-694, 2020. [PDF]
  12. Di Chen, Na Sun, Lei Hou, Rachel Kim, Jared Faith, Marianna Aslanyan, Yu Tao, Yi Zheng, Jianping Fu, Wanlu Liu, Manolis Kellis, and Amander Clark. Human primordial germ cells are specified from lineage-primed progenitors. Cell Reports, vol. 29, pp. 4568-4582, 2019. [PDF]
  13. Yi Zheng, Xufeng Xue, Yue Shao, Sicong Wang, Sajedeh Nasr Esfahani, Zida Li, Jonathon M. Muncie, Johnathon N. Lakins, Valerie M. Weaver, Deborah L. Gumucio, and Jianping Fu. Controlled modeling of human epiblast and amnion development using stem cells. Nature, vol. 573, pp. 421-425, 2019. [PDF]
  14. Nicolas Rivron, Martin Pera, Janet Rossant, Alfonso Martinez Arias, Magdalena Zernicka-Goetz, Jianping Fu, Suzanne van den Brink, Annelien Bredenoord, Wybo Dondorp, Guido de Wert, Insoo Hyun, Megan Munsie, and Rosario Isasi. Commentary: Debate ethics of embryo models from stem cells. Nature, vol. 564, pp. 183-185, 2018. [PDF]
  15. Xufeng Xue, Yubing Sun, Agnes M. Resto-Irizarry, Ye Yuan, Koh Meng Aw Yong, Yi Zheng, Shinuo Weng, Yue Shao, Yimin Chai, Lorenz Studer, and Jianping Fu. Mechanics-guided embryonic patterning of neuroectoderm tissue from human pluripotent stem cells. Nature Materials, vol. 17, pp. 633-641, 2018. [PDF | Supplemental Materials]
  16. Yue Shao, Kenichiro Taniguchi, Ryan F. Townshend, Toshio Miki, Deborah L. Gumucio, and Jianping Fu. A pluripotent stem cell-based model for post-implantation human amniotic sac development. Nature Communications, vol. 8, 208, 2017. [PDF | Supplemental Materials]
  17. Yue Shao, Kenichiro Taniguchi, Katherine Gurdziel, Ryan F. Townshend, Xufeng Xue, Koh Meng Aw Yong, Jianming Sang, Jason R. Spence, Deborah L. Gumucio, and Jianping Fu. Self-organized amniogenesis by human pluripotent stem cells in a biomimetic implantation-like niche. Nature Materials, vol. 16, pp. 419-425, 2017. [PDF | Supplemental Materials]
  18. Kenichiro Taniguchi, Yue Shao, Ryan F. Townshend, Yu-Hwai Tsai, Cynthia J. DeLong, Shawn A. Lopez, Srimonta Gayen, Andrew M. Freddo, Deming J. Chue, Dennis J. Thomas, Jason R. Spence, Benjamin Margolis, Sundeep Kalantry, Jianping Fu, K. Sue O’Shea, and Deborah L. Gumucio. Lumen formation is an intrinsic property of isolated human pluripotent stem cells. Stem Cell Reports, vol. 5, pp. 954-962, 2015. [PDF | Supplemental Materials]
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Modeling Neurodevelopment

The foundation for the anatomical and functional complexity of the vertebrate central nervous system (CNS) is laid during the development of neural tube (NT), the embryonic precursor to the CNS. Development of the NT starts from the formation of neural plate in dorsal ectoderm through the neural induction process, before its infolding into the tubular NT structure. Continuous NT development involves patterning and differentiation of distinct classes of neuronal progenitor cells located at defined positions within the NT along the anterior (A)-posterior (P) and dorsal (D)-ventral (V) axes. Neural patterning along the A-P axis establishes the main subdivisions of CNS: forebrain, midbrain, hindbrain and spinal cord. Development of human NT is a tightly regulated, genetically encoded process. Any deviation from normal NT development can result in neural tube defects and neurodevelopmental disorders and may lead to distinct neurological and psychiatric diseases later in life.

Despite the importance of NT development, we still have very limited understanding of the signaling activities and genetic networks that control human NT development. Recently, we have leveraged the developmental potential and self-organization property of human stem cells in conjunction with 2D and 3D bioengineering tools to achieve the development of patterned multicellular tissues that mimic certain aspects of early human NT development, including neural induction and D-V patterning of the NT. More recently, we have developed controllable microfluidic gradient devices to achieve the development of another human NT development model, termed microfluidic neural tube-like structure (or µNTLS). The µNTLS recapitulates some critical aspects of neural patterning in both the brain and spinal cord regions and along both A-P and D-V axes. The µNTLS is utilized for studying development of different neuronal lineages, revealing pre-patterning of axial identities of neural crest progenitors and functional roles of neuromesodermal progenitors and caudal gene CDX2 in spinal cord and trunk neural crest development. We have further developed D-V patterned, microfluidic forebrain-like structures (µFBLS) with spatially segregated dorsal and ventral regions and layered apicobasal cellular organizations that mimic human embryonic brain development in pallium and subpallium areas, respectively. Together, both µNTLS and µFBLS offer 3D lumenal tissue architectures with an in vivo-like spatiotemporal cell differentiation and organization, very promising for studying human neural development and related diseases.


Stem cell-based models of human neurulation. (left) Neurulation in human embryos. (top right) hPSC-based model of neural induction. (bottom right) hPSC-based, D-V patterned neural tube model.


Stem cell-based models of human neural patterning. (left) Neural patterning in human brain and spinal cord regions. (right) A microfluidic neural tube-like structure (µNTLS) that recapitulates regional fate patterning in both the brain and spinal cord regions and along both A-P and D-V axes.

Selected Publications:

  1. Jun Wu and Jianping Fu. Towards developing human organs via embryo models and chimeras. Cell, vol. 187, pp. 3194-3219, 2024. [PDF]
  2. Xufeng Xue, Yung Su Kim, Alfredo-Isaac Ponce-Arias, Richard O'laughlin, Robin Zhexuan Yan, Norio Kobayashi, Rami Yair Tshuva, Yu-Hwai Tsai, Shiyu Sun, Yi Zheng, Yue Liu, Frederick C.K. Wong, Azim Surani, Jason R. Spence, Hongjun Song, Guo-Li Ming, Orly Reiner, and Jianping Fu. A patterned human neural tube model using microfluidic gradients. Nature, vol. 628, pp. 391-399, 2024. [PDF | Supplemental Materials]
  3. Yue Shao, and Jianping Fu. Engineering multiscale structural orders for high-fidelity embryoids and organoids. Cell Stem Cell, vol. 29, pp. 722-743, 2022. [PDF]
  4. Yuanyuan Zheng, Xufeng Xue, Agnes M. Resto Irizarry, Zida Li, Yue Shao, Yi Zheng, Gang Zhao, and Jianping Fu. Dorsal-ventral patterned neural cyst from human pluripotent stem cells in a neurogenic niche. Science Advances, vol. 5, eaax5933, 2019. [PDF | Supplemental Materials]
  5. Xufeng Xue, Yubing Sun, Agnes M. Resto-Irizarry, Ye Yuan, Koh Meng Aw Yong, Yi Zheng, Shinuo Weng, Yue Shao, Yimin Chai, Lorenz Studer, and Jianping Fu. Mechanics-guided embryonic patterning of neuroectoderm tissue from human pluripotent stem cells. Nature Materials, vol. 17, pp. 633-641, 2018. [PDF | Supplemental Materials]
  6. Yubing Sun, Koh Meng Aw Yong, Luis G. Villa-Diaz, Xiaoli Zhang, Weiqiang Chen, Renee Philson, Shinuo Weng, Haoxing Xu, Paul H. Krebsbach, and Jianping Fu. Hippo/YAP-mediated rigidity-dependent motor neuron differentiation of human pluripotent stem cells. Nature Materials, vol. 13, pp. 599-604, 2014. [PDF | Supplemental Materials] 
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Mechanobiology of Human Pluripotent Stem Cells

Research on human pluripotent stem cells (hPSCs) has expanded rapidly over the last two decades, owing to the promise of hPSCs for regenerative medicine, disease modeling, and developmental biology studies. In our research, we have uniquely focused on a high-risk, high-payoff concept to investigate an emerging functional connection between mechanobiology and some critical questions in the field of hPSCs, including pluripotency, directed differentiation, cell reprogramming and transdifferentiation, and functional maturation. We are also exploring intracellular molecular mechanisms underlying mechanosensitive properties of hPSCs. Our research on mechanobiology of hPSCs will potentially enable drastic advances in large-scale production of hPSCs and their derivatives and contribute significantly to future cell-based regenerative therapies and disease modeling. So far, our research has unambiguously unraveled the mechanosensitive properties of hPSCs and their roles in directed neural differentiation and subtype specification. Our mechanistic work has further led to the discovery of an intervened regulatory network emerged from converging and reinforcing signal integration of TGF-β, WNT, Hippo, Rho-GTPase, and the actomyosin cytoskeleton that forms a molecular framework required for contextual, integrated responses of hPSCs.


Micro/nanoengineered ex vivo stem cell niche.

Selected Publications:

  1. Yue Liu, Xufeng Xue, Shiyu Sun, Norio Kobayashi, Yung Su Kim, and Jianping Fu. Morphogenesis beyond in vivo. Nature Reviews Physics, vol. 6, pp. 28-44, 2024. [PDF]
  2. Feng Lin, Xia Li, Shiyu Sun, Zhongyi Li, Jianbo Bai, Lin Song, Bo Li, Jianping Fu, and Yue Shao. Mechanically enhanced biogenesis of gut spheroids with instability-driven morphomechanics. Nature Communications, vol. 14, 6016, 2023. [PDF | Supplemental Materials]
  3. Yue Shao, and Jianping Fu. Engineering multiscale structural orders for high-fidelity embryoids and organoids. Cell Stem Cell, vol. 29, pp. 722-743, 2022. [PDF]
  4. Jonathon M. Muncie, Nadia M.E. Ayad, Johnathon N. Lakins, Xufeng Xue, Jianping Fu, and Valerie M. Weaver. Mechanical tension promotes formation of gastrulation-like nodes and patterns mesoderm specification in human embryonic stem cells. Developmental Cell, vol. 55, pp. 679-694, 2020. [PDF]
  5. Xufeng Xue, Yubing Sun, Agnes M. Resto-Irizarry, Ye Yuan, Koh Meng Aw Yong, Yi Zheng, Shinuo Weng, Yue Shao, Yimin Chai, Lorenz Studer, and Jianping Fu. Mechanics-guided embryonic patterning of neuroectoderm tissue from human pluripotent stem cells. Nature Materials, vol. 17, pp. 633-641, 2018. [PDF | Supplemental Materials]
  6. Yue Shao, Kenichiro Taniguchi, Katherine Gurdziel, Ryan F. Townshend, Xufeng Xue, Koh Meng Aw Yong, Jianming Sang, Jason R. Spence, Deborah L. Gumucio, and Jianping Fu. Self-organized amniogenesis by human pluripotent stem cells in a biomimetic implantation-like niche. Nature Materials, vol. 16, pp. 419-425, 2017. [PDF | Supplemental Materials]
  7. Yubing Sun and Jianping Fu. Harnessing mechanobiology of human pluripotent stem cells for regenerative medicine. ACS Chemical Neuroscience, vol. 5, pp. 621-623, 2014. [PDF]
  8. Yubing Sun, Koh Meng Aw Yong, Luis G. Villa-Diaz, Xiaoli Zhang, Weiqiang Chen, Renee Philson, Shinuo Weng, Haoxing Xu, Paul H. Krebsbach, and Jianping Fu. Hippo/YAP-mediated rigidity-dependent motor neuron differentiation of human pluripotent stem cells. Nature Materials, vol. 13, pp. 599-604, 2014. [PDF | Supplemental Materials] 
  9. Yubing Sun and Jianping Fu. Mechanobiology: A new frontier for human pluripotent stem cells. Integrative Biology, vol. 5, pp. 450-457, 2013. [PDF]
  10. Yubing Sun, Luis G Villa-Diaz, Raymond Hiu-Wai Lam, Weiqiang Chen, Pasul H. Krebsbach, and Jianping Fu. Matrix mechanics regulates fate decisions of human embryonic stem cells. PLoS ONE, vol. 7, e37178, 2012. [PDF]
  11. Weiqiang Chen, Luis G Villa-Diaz, Yubing Sun, Shinuo Weng, Raymond Hiu-Wai Lam, Lin Han, Rong Fan, Paul H. Krebsbach, and Jianping Fu. Nanotopography influences adhesion, spreading, and self-renewal of human embryonic stem cells. ACS Nano, vol. 6, pp. 4094-4103, 2012. [PDF | supplemental materials]
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Cell Mechanics and Mechanotransduction

External forces and matrix mechanics play a key role in the regulation of cell function. Cells sense and response to external forces and changes in matrix mechanics by modulating their endogenous cytoskeleton (CSK) contractility, balanced by external forces or resistant forces generated by the deformation of the extracellular matrix (ECM). Dysregulation of the tensional homeostasis in cells contributes to atherosclerosis, osteoarthritis and osteoporosis, and cancer. To aid in the mechanistic investigation of mechanotransduction, we have developed different micromechanical tools that allow for quantitative controls and real-time measurements of mechanical stimuli and cellular biomechanical responses. Our micromechanical tools are particularly useful for investigations of mechanotransduction centering on the ECM-integrin-CSK signaling axis to generate quantitative descriptions of the functional relations between matrix mechanics, external forces, CSK contractility, cell stiffness, and adhesion signaling and morphogenesis.


Micromechanical tools for precise control and measurement of mechanical stimuli and responses.

Selected Publications:

  1. Dennis W. Zhou, Marc A. Fernández-Yagüe, Elijah N. Holland, Andrés F. García, Nicolas S. Castro, Eric B. O’Neill, Jeroen E.G. Eyckmans, Christopher S. Chen, Jianping Fu, David D. Schlaepfer, and Andrés J. García. Force-FAK signaling coupling at individual focal adhesions coordinates mechanosensing and microtissue repair. Nature Communications, vol. 12, 2359, 2021. [PDF]
  2. Jonathon M. Muncie, Nadia M.E. Ayad, Johnathon N. Lakins, Xufeng Xue, Jianping Fu, and Valerie M. Weaver. Mechanical tension promotes formation of gastrulation-like nodes and patterns mesoderm specification in human embryonic stem cells. Developmental Cell, vol. 55, pp. 679-694, 2020. [PDF]
  3. Shinuo Weng, Yue Shao, Weiqiang Chen, and Jianping Fu. Mechanosensitive subcellular rheostasis drives emergent single-cell mechanical homeostasis. Nature Materials, vol. 15, pp. 961-967, 2016. [PDF | Supplemental Materials]
  4. Di Chen, Yubing Sun, Madhu S. R. Gudur, Yising Hsiao, Ziqi Wu, Jianping Fu, and Cheri X. Deng. Two bubble acoustic tweezing cytometry for biomechanical probing and stimulation of cells. Biophysical Journal, vol. 108, pp. 32-42, 2015. [PDF]
  5. Yue Shao, Jennifer M. Mann, Weiqiang Chen, and Jianping Fu. Global architecture of F-actin cytoskeleton regulates cell shape-dependent endothelial mechanotransduction. Integrative Biology, vol. 6, pp. 300-311, 2014. [PDF | Supplemental Materials]
  6. Zhenzhen Fan, Yubing Sun, Di Chen, Donald Tay, Weiqiang Chen, Cheri X. Deng, and Jianping Fu. Acoustic tweezing cytometry for live-cell subcellular control of intracellular cytoskeleton contractility. Scientific Reports, vol. 3, 2176, 2013. [PDF | Supplemental Materials]
  7. Raymond Hiu-Wai Lam, Shinuo Weng, Wei Lu, and Jianping Fu. Live-cell subcellular measurement of cell stiffness using a microengineered stretchable micropost array membrane. Integrative Biology, vol. 4, pp. 1289-1298, 2012. [PDF]
  8. Raymond Hiu-Wai Lam, Yubing Sun, Weiqiang Chen, and Jianping Fu. Elastomeric microposts integrated into microfluidics for flow-mediated endothelial mechanotransduction analysis. Lab on a Chip, vol. 12, pp. 1865-1873, 2012. [PDF | supplemental materials]
  9. Jennifer M. Mann, Raymond Hiu-Wai Lam, Shinuo Weng, Yubing Sun, and Jianping Fu. A silicone-based stretchable micropost array membrane for monitoring live-cell subcellular cytoskeletal response. Lab on a Chip, vol. 12, pp. 731-740, 2012. [PDF | supplemental materials]
  10. Shang-You Tee, Jianping Fu, Christopher S. Chen, and Paul A. Janmey. Cell shape and substrate rigidity both regulate cell stiffness. Biophysical Journal, vol. 100, pp. L25-27, Mar. 2011. [PDF | supplemental materials]
  11. Michael T. Yang, Jianping Fu, Yang-Kao Wang, Ravi A. Desai, and Christopher S. Chen. Assaying stem cell mechanobiology on microfabricated elastomeric substrates with geometrically modulated rigidity. Nature Protocols, vol. 6, pp. 187-213, 2011. [PDF]
  12. Jianping Fu, Yang-Kao Wang, Michael T. Yang, Ravi A. Desai, Xiang Yu, Zhijun Liu, and Christopher S. Chen. Mechanical regulation of stem cell function using geometrically modulated elastomeric substrates. Nature Methods, vol. 7, pp.733-736, 2010. [PDF | supplemental materials]
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