SfN’s 48th annual meeting, Neuroscience 2018, is the world’s largest neuroscience conference for scientists and physicians devoted to understanding the brain and nervous system.
Meet the Author
To celebrate the upcoming release of iCell® Microglia, join Fujifilm Cellular Dynamics in welcoming Edsel Abud, the lead author of:
Join us at the FCDI booth at Neuroscience 2018 to:
- Discuss your research applications
- Explore the benefits of iPSC-derived microglia
- Be eligible for free vials of iCell Microglia to jump start your research (quantities limited)
Fujifilm Cellular Dynamics Poster Presentations
The Generation and Functional Characterization of Human Microglia from Episomally Reprogrammed iCell Hematopoietic Progenitor Cells
Microglia are immune-competent cells residing within the brain that play a critical role in maintaining immunological balance for normal brain function. As phagocytic cells, they are activated following clearance of pathogens and secreted proteins and respond by either exacerbating or dampening a pro-inflammatory environment. Primary human microglia are difficult to acquire and stably culture in vitro. We generated and characterized functional human induced pluripotent stem cell-derived microglia (iCell Microglia) from episomally reprogrammed iCell Hematopoietic Progenitor Cells (proprietary technology) under defined conditions based on technology developed by the Blurton-Jones laboratory exclusively licensed to FCDI from the University of California-Irvine. iCell Microglia express CD45, CD11b and CD33, and consistent with a microglial phenotype, they also express PU.1, CX3CR1, IBA, TREM-2 and P2RY12. The purity for all these markers is greater than 80% and the retain the expression of all these markers post cryopreservation. iCell Microglia were able to phagocytose opsonized bacteria and fibrillar Aβ within a span of 2-60hrs and reveal a ramified morphology when treated with 5µM Thiazovivin. Additionally, iCell Microglia secrete cytokines and chemokines including TNFα, IL-8, IL-10, CCL2, CCL4, CCL3, CCL4, CXCL10, CXCL11, CXCl1, CXCl2 and CXCL10 when stimulated with LPS and interferon gamma. iCell Microglia will serve as a great tool for disease modeling and drug testing for neuroscience research.
675.20 / QQ8 | Wednesday, Nov. 7, 2018 | 11:00 am – noon | Halls B-H
The ability to produce motor neurons from induced pluripotent stem cells (iPSCs) provides the means to model several neuromuscular disorders using human cells. It is now possible to explore human-specific drug screening platforms with prospective treatment possibilities using iPS-derived motor neurons for disorders lacking therapeutic options such as amyotrophic lateral sclerosis (ALS). Here we demonstrate the generation of iPSc-derived motor neurons using an optimized differentiation protocol generating a population with greater than 60% purity based on Isl1/2 and Tuj1 positive staining. Furthermore, we used this cell line to generate a TDP-43 line, equipping researchers with this valuable tool for ALS prognostics and treatment. We characterize our cells and co-cultured them with human skeletal muscle, capable of forming neuromuscular junctions in vitro. We are now exploring co-culture conditions with human iPSC-derived skeletal muscle based on our published protocol (Caron et al., 2016). Our media-based differentiation method efficiently generates contractile and fused myotubes without the need for any cell sorting or myogenic gene overexpression. Combined with our motor neurons, this co-culture platform will enable ALS drug discovery and facilitate complex disease modeling of other neuromuscular disorders.
755.16 / DD18 | Wednesday, Nov. 7 | 4:00 – 5:00 pm | Halls B-H
Superior Survival, Innervation, and Functional Recovery by Day 17 Differentiated Dopamine Stem Cells in a Rat Model of Parkinson’s Disease
As the second most common neurodegenerative disease, Parkinson’s disease (PD) presents an immense burden for the aging population. PD is a progressive neurodegenerative disease characterized by the loss of dopamine neurons in the substantia nigra subsequent to the loss of striatal dopaminergic tone. The current gold standard treatment for PD is limited to oral levodopa which eventually can lead to the development of motor fluctuation and dyskinesias. Early work with fetal and embryonic tissues suggests that a cell replacement therapy may offer relief for motor deficits associated with PD. However, these tissue sources are associated with ethical and logistical concerns.
Previously, we have shown that cryopreserved human induced pluripotent stem cells (hiPSC) reprogrammed to post-mitotic midbrain dopamine (mDA) neurons survive engraftment in the rat striatum with low levels of proliferation. Further, when striatally transplanted following a 6-hydroxydopamine lesion to the medial forebrain bundle, hiPSC-mDA neurons reverse motor asymmetry as measured by amphetamine-induced rotations (Wakeman et al., 2017).
In the present study we tested the in vivo efficacy of hiPSC-mDA neurons and progenitor cells after transitioning the differentiation protocol to a cell therapy-compatible manufacturing process. 4.5x105cells were unilaterally transplanted to the 6-hydroxydopamine-lesioned striatum of athymic nude rats. Here we show that hiPSC-mDA neurons progenitors cryopreserved at day 17 (D17), D24, and D37 of the differentiation process and iCell DopaNeurons survive and project tyrosine hydroxylase-immunoreactive processes into the host parenchyma. Animals that received grafts of D17 and D24 progenitor cells showed fully reversed amphetamine-induced rotations by 6 months post-transplant, with complete reversal seen by 4 months post-grafting in animals treated with D17 progenitors. In addition, the D17 progenitors exhibited superior mDA neuron and innervation of the striatum. We found low levels of proliferating cells and no evidence of teratoma formation within the grafts as indicated by human specific Ki-67 staining and did not observe cell migration or aberrant outgrowth, demonstrating a safety profile amenable to translation to the clinic. Ongoing studies will seek to determine optimal dosing to inform future human trials.