Neural tissue engineering using embryonic and induced pluripotent stem cells

With the recent start of the first clinical trial evaluating a human embryonic stem cell-derived therapy for the treatment of acute spinal cord injury, it is important to review the current literature examining the use of embryonic stem cells for neural tissue engineering applications with a focus on diseases and disorders that affect the central nervous system. Embryonic stem cells exhibit pluripotency and thus can differentiate into any cell type found in the body, including those found in the nervous system. A range of studies have investigated how to direct the differentiation of embryonic cells into specific neural phenotypes using a variety of cues to achieve the goal of replacing diseased or damaged neural tissue. Additionally, the recent development of induced pluripotent stem cells provides an intriguing alternative to the use of human embryonic stem cell lines for these applications. This review will discuss relevant studies that have used embryonic stem cells to replicate the tissue found in the central nervous system as well as evaluate the potential of induced pluripotent stem cells for the aforementioned applications

Transdifferentiating Astrocytes Into Neurons Using ASCL1 Functionalized With a Novel Intracellular Protein Delivery Technology

Cellular transdifferentiation changes mature cells from one phenotype into another by altering their gene expression patterns. Manipulating expression of transcription factors, proteins that bind to DNA promoter regions, regulates the levels of key developmental genes. Viral delivery of transcription factors can efficiently reprogram somatic cells, but this method possesses undesirable side effects, including mutations leading to oncogenesis. Using protein transduction domains (PTDs) fused to transcription factors to deliver exogenous transcription factors serves as an alternative strategy that avoids the issues associated with DNA integration into the host genome. However, lysosomal degradation and inefficient nuclear localization pose significant barriers when performing PTD-mediated reprogramming. Here, we investigate a novel PTD by placing a secretion signal sequence next to a cleavage inhibition sequence at the end of the target transcription factor–achaete scute homolog 1 (ASCL1), a powerful regulator of neurogenesis, resulting in superior stability and nuclear localization. A fusion protein consisting of the amino acid sequence of ASCL1 transcription factor with this novel PTD added can transdifferentiate cerebral cortex astrocytes into neurons. Additionally, we show that the synergistic action of certain small molecules improves the efficiency of the transdifferentiation process. This study serves as the first step toward developing a clinically relevant in vivo transdifferentiation strategy for converting astrocytes into neurons

Advancements in Canadian biomaterials research in neurotraumatic diagnosis and therapies

Development of biomaterials for the diagnosis and treatment of neurotraumatic ailments has been significantly advanced with our deepened knowledge of the pathophysiology of neurotrauma. Canadian research in the fields of biomaterial-based contrast agents, non-invasive axonal tracing, non-invasive scaffold imaging, scaffold patterning, 3D printed scaffolds, and drug delivery are conquering barriers to patient diagnosis and treatment for traumatic injuries to the nervous system. This review highlights some of the highly interdisciplinary Canadian research in biomaterials with a focus on neurotrauma applications

Development of a Low Bias Method for Characterizing Viral Populations Using Next Generation Sequencing Technology

Background: With an estimated 38 million people worldwide currently infected with human immunodeficiency virus (HIV), and an additional 4.1 million people becoming infected each year, it is important to understand how this virus mutates and develops resistance in order to design successful therapies. Methodology/Principal Findings: We report a novel experimental method for amplifying full-length HIV genomes without the use of sequence-specific primers for high throughput DNA sequencing, followed by assembly of full length viral genome sequences from the resulting large dataset. Illumina was chosen for sequencing due to its ability to provide greater coverage of the HIV genome compared to prior methods, allowing for more comprehensive characterization of the heterogeneity present in the HIV samples analyzed. Our novel amplification method in combination with Illumina sequencing was used to analyze two HIV populations: a homogenous HIV population based on the canonical NL4-3 strain and a heterogeneous viral population obtained from a HIV patient's infected T cells. In addition, the resulting sequence was analyzed using a new computational approach to obtain a consensus sequence and several metrics of diversity. Significance: This study demonstrates how a lower bias amplification method in combination with next generation DNA sequencing provides in-depth, complete coverage of the HIV genome, enabling a stronger characterization of the quasispecies present in a clinically relevant HIV population as well as future study of how HIV mutates in response to a selective pressure

3-D Bioprinting of Neural Tissue for Applications in Cell Therapy and Drug Screening

Neurodegenerative diseases affect millions of individuals in North America and cost the health-care industry billions of dollars for treatment. Current treatment options for degenerative diseases focus on physical rehabilitation or drug therapies, which temporarily mask the effects of cell damage, but quickly lose their efficacy. Cell therapies for the central nervous system remain an untapped market due to the complexity involved in growing neural tissues, controlling their differentiation, and protecting them from the hostile environment they meet upon implantation. Designing tissue constructs for the discovery of better drug treatments are also limited due to the resolution needed for an accurate cellular representation of the brain, in addition to being expensive and difficult to translate to biocompatible materials. 3-D printing offers a streamlined solution for engineering brain tissue for drug discovery or, in the future, for implantation. New microfluidic and bioplotting devices offer increased resolution, little impact on cell viability and have been tested with several bioink materials including fibrin, collagen, hyaluronic acid, poly(caprolactone), and poly(ethylene glycol). This review details current efforts at bioprinting neural tissue and highlights promising avenues for future work

Parametric Studies of Melt Electrospinning Poly ?(caprolactone) Fibers for Tissue Engineering Applications

The process of electrospinning has received remarkable amount of attention as this technique can be used to fabricate suitable scaffolds for cells. In order to control morphology of scaffold, parametric studies are necessary in customized melt electrospinning. In this study, we demonstrate that fiber diameter could be controlled by customized nozzle and parameters which are temperature, voltage, and distance influence to fiber diameter. Moreover, we culture and seed murine CE3 stem cells derived from D3 embryonic stem cell line on fabricated scaffolds. Overall, scaffold controlled by parameter studies holds a promising strategy for superb cell attachment and proliferation

Utilizing Additive Manufacturing to Produce Organ Mimics and Imaging Phantoms

The complex geometries and material properties necessary for generating accurate organ mimics require new procedures and methods to fully utilize current technologies. The increased accessibility of 3D printers, along with more specialized bioprinters, allow the creation of highly tunable models of various body parts. Three-dimensional printing can reduce lead-time on custom parts, produce structures based on imaging data in patients, and generate a test bench for novel surgical methods. This technical note will cover three unique case studes and offer insights for how 3D printing can be used for lab research. Each case follows a unique design process in comparison to traditional manufacturing workflows as they required significantly more iterative design. The strengths of different printing technologies, design choices, and structural/chemical requirements all influence the design process. Utilization of in-house manufacturing allows for greater flexibility and lower lead-times for novel research applications. Detailed discussions of these design processes will help reduce some of the major barriers to entry for these technologies and provide options for researchers working in the field

3D Printing Breast Tissue Models : A Review of Past Work and Directions for Future Work

Breast cancer often results in the removal of the breast, creating a need for replacement tissue. Tissue engineering offers the promise of generating such replacements by combining cells with biomaterial scaffolds and serves as an attractive potential alternative to current surgical repair methods. Such engineered tissues can also serve as important tools for drug screening and provide in vitro models for analysis. 3D bioprinting serves as an exciting technology with significant implications and applications in the field of tissue engineering. Here we review the work that has been undertaken in hopes of generating the recognized in-demand replacement breast tissue using different types of bioprinting. We then offer suggestions for future work needed to advance this field for both in vitro and in vivo applications.Medicine, Faculty ofNon UBCUrologic Sciences, Department ofReviewedFacult