Synthetic Embryology: Post-Implantation Development

Synthetic Embryology: Neurodevelopment

Mechanobiology of Human Pluripotent Stem Cells

Cell Mechanics and Mechanotransduction

 

Synthetic Embryology: Post-Implantation Development

Human post-implantation development remains mysterious and difficult to study. Recent advances in the development of human embryo models (or embryoids) from human pluripotent stem cells (hPSCs) have sparked great interest in using such models to advance human embryology and regenerative and reproductive medicine. In this research, we have leveraged the developmental potential and self-organizing property of hPSCs in conjunction with biomimetic culture systems to develop different embryoid systems to study the early post-implantation human development. Specifically, we have successfully developed the first hPSC-based embryoid system that recapitulates successive embryogenic events during the early post-implantation human development, including pro-amniotic cavity formation, amnion-epiblast patterning, specification of primordial germ cells, and the onset of gastrulation. Together, our studies have successfully established faithful experimental models of human development and provided insights into previously inaccessible phases of the post-implantation human development.


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. 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]
  2. 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]
  3. 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]
  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. 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]
  6. 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]
  7. 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]
  8. 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]
  9. 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]
  10. 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]
  11. 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]

Synthetic Embryology: 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 a tubular structure enclosing a central fluid-filled lumen (the neural canal). Continuous development of the NT 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. Neuronal patterning along the A-P axis establishes the main subdivisions of CNS: forebrain, midbrain, hindbrain and spinal cord. The development of human NT is a tightly regulated, genetically encoded process. Any deviation from the normal NT developmental program 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 neuronal patterning. Recently, we have leveraged the developmental potential and self-organization property of human pluripotent stem cells (hPSCs) 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 DV patterning of the NT. These stem cell-derived models offer promising trackable systems to study neural development and diseases.


Stem cell-based models of human neural system development. (left) Neurulation in vertebrate embryos. (top right) hPSC-based model of neural induction. (bottom right) hPSC-based, dorsal-ventral patterned neural tube model.

Selected Publications:

  1. 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]
  2. 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]
  3. 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

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. 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]
  2. 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]
  3. 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]
  4. 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]
  5. 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
  6. Yubing Sun and Jianping Fu. Mechanobiology: A new frontier for human pluripotent stem cells. Integrative Biology, vol. 5, pp. 450-457, 2013. [PDF]
  7. 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]
  8. 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]

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]