Biohybridized Neural Interface Microsystems

Living Neuro-Prosthetic Interfaces

The Cullen Lab blends neural tissue engineering techniques and micro-electrical technology to create “biohybrid” interface microsystems. These living biohybridized neural-electrical interface microsystems are being investigated for functional integration with the nervous system. With this design, host axonal ingrowth and synaptic integration may occur with the living component, potentially exploiting a more natural integration with the non-organic interface.  In the case of driving a prosthetic limb, the neural interface may occur with the peripheral nervous system rather than placing a device into the otherwise non-injured brain or spinal cord.  In addition, this neural interface occurs at the point of final motor output and primary sensory input, thus leveraging the exquisite processing power of the brain and spinal cord rather than straining to decipher it.  However, peripheral nerve axonal require a living target for innervation, hence the necessity of our living biohybrid neural interfaces.  Overall, these tissue engineered 3-D neural interfce microsystems may significantly advance regeneration or device-based deficit mitigation in the nervous system.



Biohybridized neural interface microsystems formed around 3-D electrodes create unique microsystems that are powerful platforms for enhancing the interface with the nervous system.  (a) Cells can be grown in vitro on electro-conductive microfibers and encapsulated with hydrogel or (b) the microfibers can be incorporated with the axonal constructs prior to transplantation.

neurons and azons


Controlled neuronal adhesion to conductive polymer microfibers.  Confocal reconstructions of neuronal cultures plated on microfibers immunolabeled at 7 days in vitro for MAP-2 (green) and tau (fiber locations denoted by dashed lines). By controlling the relative electrostatic surface charge of the microfiber and the substrate, adhesion to the microfiber was increased.  (a) Low-density adhesion on the microfibers resulted in the axonal projections to the substrate.  (b) Robust neuronal adhesion resulted in neuronal somata and axonal containment on the microfiber. Scale bar = 200 µm.




Neuronal encapsulation on microfibers.  For future transplantation, removal from culture while maintaining neuronal network integrity and viability is necessary.  In order to demonstrate this using hydrogel encapsulation, neurons were plated on collagen-coated conductive polymer microfibers and, at 6-9 days in vitro, encapsulated using 0.5-1.0% agarose.  (a-c) Representative fluorescent confocal reconstructions of encapsulated neuronal cultures on microfibers stained to discriminate live cells (green) from the nuclei of dead cells (red) (scale bar = 200 µm). (a) The encapsulation process did not reduce the cell density or the cell viability versus non-encapsulated controls.  Increased magnification of regions of interest from showing (b) a cluster of neuronal somata and (c) a neurite-rich segment following encapsulation (scale bars = 50 µm).


Advanced Microsystems In Vitro

These techniques are also useful to enhance the capabilities of investigational platforms in vitro.  Here, interfacing novel 3-D neural cellular constructs with micro-fluidic and/or micro-electrical systems has created biohybridized platforms, providing unprecendented 3-D microenvironmental control and allowing non–invasive probing and manipulation of cultured neural cells. This effort was selected as one of the Highlights of Neural Engineering entitled “Microfluidic Engineered High Cell Density 3-D Neural Cultures”. Currently, the Cullen Lab is applying these 3-D neural cellular constructs as powerful investigational platforms for the study of basic neurobiology, network neurophysiology, injury/disease mechanisms, pharmacological screening, or test-beds for cell replacement therapies.

advanced microsystems


Biohybridized neural interfaces. (a-b) Concept: 3-D neural cell cultures formed around 3-D micro-towers to create biohybridized neural interface microsystems. (c-d) Live neurons (green fluorescent protein; GFP+) forming 3-D networks around micro-towers in culture. (c) At 1 DIV, neurons are homogenously distributed throughout the matrix (>500 µm thick); (d) However, by 13 DIV, there was increased cell density around the towers and the culture thickness had decreased to approximately 300 µm. (e) Live (green) and dead (red) neural cells at 23 DIV, demonstrating viable neural networks adhering to the micro-towers.  (f-g) Immunocytochemistry labeling astrocytes (GFAP+, red) with GFP+ neurons. Neurons and astrocytes were in intimate contact with each other and the micro-towers in the bottom 300 mm of the cultures. (h) In some cases, neural cells and processes had coalesced into tracts spanning neural populations on micro-towers (top 200 µm shown here). With micro-electrical capability, these microsystems may serve as powerful in vitro platforms to continuously monitor the electrical activity and dynamics of “small world” neuronal networks in 3-D matrices.



Contact Information

D. Kacy Cullen, Ph.D
105E Hayden Hall, 3320 Smith Walk
Philadelphia, PA 19104

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News and Events

  • Dr. John O’Donnell, a postdoctoral fellow in the Cullen Lab, was awarded a prestigious NRSA (F32) Training Fellowship through the NIH. His project is titled “Modeling Disorders of Consciousness”. Way to go John!
  • The Cullen Lab and collaborators had a poster selected as one of the "Blue Ribbon Highlights" at the International Congress of Parkinson's Disease and Movement Disorders in Vancouver, Canada. The poster was titled “Tissue Engineered Nigrostriatal Pathway for Treatment of Parkinson’s Disease”. A write-up about the project in Neurology Today can be found here.
  • Dr. Cullen and collaborators received a major 4-year Merit Review Award from the Department of Veterans Affairs to advance their project “Transplantable Micro-Tissue Engineered Neural Networks to Restore the Nigrostriatal Pathway in Parkinson's Disease”. This exciting project applies cutting-edge tissue engineering and regenerative medicine techniques to build the full nigrostriatal pathway in vitro using human induced pluripotent stem cells, with efficacy testing of these human-scale constructs in a large animal model of Parkinson’s disease. Success of this project over the next several years will enable clinical trials based on precision medicine– rebuilding complex brain networks using a patients own cells and implanting these fully preformed constructs to replace vulnerable pathways lost due to neurodegenerative disease. The co-investigators on this project are Dr. Isaac Chen, Dr. John Wolf, and Dr. John Duda. Way to go team!
  • Dr. Cullen was promoted to Associate Professor at the University of Pennsylvania. Congratulations Dr. Cullen!
  • Congratulations to Laura Struzyna, a Bioengineering Doctoral Candidate in the Cullen Lab, whose winning entry for Penn's "Art in Science" competition was featured on the NIH Director's Blog!
  • The Cullen Lab and collaborators at Penn (Drs. Isaac Chen, Douglas Smith, and John Wolf) and Rutgers University (Drs. Joachim Kohn and Hilton Kaplan) were awarded a 3-year, $2.1 million dollar grant from the U.S. Dept. of Defense on "Tissue Engineering Strategies to Maintain Distal Target Efficacy and Promote Full Functional Recovery Following Major Peripheral Nerve Injury".
  • The Cullen Lab and collaborators had two abstracts selected as “Hot Topics” (top 2% of submissions) for presentation in a platform session at the World Parkinson’s Congress meeting in Portland, OR. Carolyn Keating, a Neuroscience doctoral candidate, presented "Diffuse Brain Injury in Swine Causes Plasmalemmal Dysruption and α-Synuclein Over-Expression in the Substantia Nigra". Dr. John Duda presented "Can Living Micro-Tissue Engineered Axonal Tracts Reconstruct the Nigrostriatal Pathway in PD?"
  • Dr. Cullen had the honor of being selected as one of "3 People to Watch in Neuroscience in 2016" by STAT News (Health, Medicine, and Biotech publication):
  • Congratulations to Laura Struzyna, a Bioengineering Doctoral Candidate in the Cullen Lab, who won second place in the Perelman School of Medicine 2016 "Art in Science" Competition. The winning image is shown here.
  • Penn and the Journal of Neural Engineering put out a press release for our most recent manuscript describing the Cullen Lab's novel micro-tissue engineered neural networks to restore brain circuitry. Link:
  • Carla Winter, and undergraduate researcher in the Cullen Lab for the past 3 years, was selected as a George J. Mitchell Scholar (one of only 12 in the U.S.): is very prestigious and well-deserved honor. Way to go Carla!
  • Dr. Cullen had the honor of giving a TED-style talk at Penn’s Annual Neuroscience Public Lecture Series on Neuroengineering: Where Biology Meets Technology. Dr. Cullen discussed the lab’s pioneering work on "Restoring Nervous System Structure and Function With Tissue Engineered 'Living Scaffolds'” (YouTube video:
  • The Cullen Lab and collaborators Dr. John Wolf, Dr. Misha Serruya, Dr. Brain Litt, Dr. Reuben Kraft and Dr. Isaac Chen were awarded a 3-year, $2 million dollar cooperative agreement (U01) from the NIH to further pioneer "Biological 'Living Electrodes' Using Tissue Engineered Axonal Tracts to Probe and Modulate the CNS".
  • The Cullen Lab received a 2-year grant from the U.S. Dept. of Defense to advance "Novel Tissue-Engineered Nerve Grafts for Repair of Currently Untreatable Peripheral Nerve Injury".
  • Bioengineering Ph.D. Candidate, Kritika Katiyar, was selected for a prestigious NRSA F31 Graduate Research Fellowship through the National Institutes of Health based on her proposal for "Molecular Mediators of Axon-Facilitated Axon Regeneration."
  • Dayo Adewole, a Bioengineering Ph.D. student in the Cullen Lab, was awarded a 3-year Graduate Research Fellowship through the National Science Foundation to develop next-generation neuroprosthetic interfaces. Way to go Dayo!
  • The Cullen Lab and collaborators Dr. John Duda, Dr. Isaac Chen, and Dr. John Wolf were awarded a 2-year grant from the Michael J. Fox Foundation for the project "Restoring the Nigrostriatal Pathway with Living Micro-Tissue Engineered Axonal Tracts".

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