Cedars-sinai Medical Center

Emulate, Inc.

Wisconsin Alumni Research Foundation

Other

Clive N Svendsen

Dhruv Sareen

Daniel Levner

Christopher David Hinojosa

Geraldine A Hamilton

Norman Wen

Jacob Fraser

S Jordan Kerns

Samuel Sances

Gad Vatine

Robert N Barrett

Carol Lucchesi

Michael Workman

Loren A Ornelas

Uthra Rajamani

Carolina Lucchesi

Magdalena Kasendra

Jefferson Puerta

Stephen R Targan

Alexander Laperle

Nur Yucer

Pablo Avalos

Doniel Drazin

Eric V Shusta

Chris Hinojosa

Stephan R Targan

Sean Paul Palecek

Mark Sasha Drlik

Michael John Baker

Joshua Breunig

Beth Y Karlan

Christian Weidenfeller

Ritchie Ho

Allison Ebert

Scott G Canfield

Tilly Jenkins Vogel

Aslam Akhtar

SJordan Kerns

Marie Holzapfel

Samuel Sanees

Sam Sances

Jordan Kerns

Magdelena Kasendra

Gad D Vatine

Alexis J Kerl

The invention relates to culturing brain endothelial cells, and optionally astrocytes and neurons in a fluidic device under conditions whereby the cells mimic the structure and function of the blood brain barrier. Culture of such cells in a microfluidic device, whether alone or in combination with other cells, drives maturation and/or differentiation further than existing systems.

Microfluidic model of the blood brain barrier

Organs-on-chips are microfluidic devices for culturing living cells in micrometer sized chambers in order to model physiological functions of tissues and organs. Engineered patterning and continuous fluid flow in these devices has allowed culturing of intestinal cells bearing physiologically relevant features and sustained exposure to bacteria while maintaining cellular viability, thereby allowing study of inflammatory bowl diseases. However, existing intestinal cells do not possess all physiologically relevant subtypes, do not possess the repertoire of genetic variations, or allow for study of other important cellular actors such as immune cells. Use of iPSC-derived epithelium, including IBD patient-specific cells, allows for superior disease modeling by capturing the multi-faceted nature of the disease.

Systems and methods for growth of intestinal cells in microfluidic devices

Delivery of glial cell line-derived neurotrophic factor (GDNF) has provided benefits to Parkinsonian patients and is currently being tested in a Phase 1/2a clinical trial for ALS patients. However, chronic trophic factor delivery prohibits dose adjustment or shut off in the event of side effects. To address this, the Inventors engineered a stably integrating, third-generation doxycycline-regulated vector, allowing inducible and reversible expression of a therapeutic molecule Human iPSC-derived neural progenitors were stably transfected with the vector, expanded and transplanted into the adult mouse brain. The Inventors observed that the addition and withdrawal of doxycycline led to GDNF expression that could be induced and reversed multiple times, demonstrating that doxycycline can penetrate the graft and regulate transgene expression in vivo. The Inventors' findings provide a proof of concept for combining gene and stem cell therapy for effective modulation of ectopic protein expression in transplanted cells.

Methods and compositions for inducible expression of neurotrophic factors

Described herein is a microphysiological system for models of disease. Specifically, induced pluripotent stem cells (iPSCs) and iPSC-derived cells, including those obtained from disease patients, are seeded onto microfluidic 'chip' devices to study cellular development and disease pathogenesis. Herein, neurodegenerative disease modeling, including Parkinson's Disease (PD) is shown to reproduce key PD pathology in a vascularized human model that contains neurons relating to PD pathology. Such compositions and methods are used for research for PD biomarkers, patient screening for PD risk assessment, and therapeutic discovery and testing. A panel of biomarkers are generated through analysis of living PD-chips by neural activity, whole transcriptomic, proteomic, and metabolomic analysis, and functional enzyme tests of media and tissue. Introducing therapeutics through a vasculature channel, coupled with blood brain barrier penetration studies can be assessed for efficacy in the human neural cells present in the PD-Chip.

Human pluripotent stem cell derived neurodegenerative disease models on a microfluidic chip

Development of spinal cord on a microfluidic chip

Type 2 diabetes (T2D) is a clinical syndrome caused by insufficient insulin secretion for insulin requirements. described herein are compositions and methods for microphysiological MPS models of disease (MODs) for diabetes. These platforms allow one to compare the effect of chronic β-cell stimulation in the presence and absence of patient specific immune cells in IPSC-derived islets from each group. Additionally, one can reproduce the T2D β-cell phenotype, using islets-on-chips will also be exposed to gluco-lipotoxicity. Likewise, skeletal muscle-on-chips are exposed to patient specific activated immune cells, variable motor neuron innervation and lipids characteristic of T2D.

Methods of use of islet cells

Described herein is functionalized glass allowing for robust attachment of extracellular matrix proteins (ECM) withstanding extended culturing periods. By first treating glass with a sulfur silane reagent, the treated glass can be activated via an amine-sulfur linker, after which ECM proteins are attached to the linker. The Inventors observed that this glass treatment combination (sulfur silane-linker-ECM) resisted degradation when compared to conventional surface coatings, such as poly-L-orthinine coated glass.

L-glass: a novel functionalization method for covalently attaching ECM protein to optical glass

Induced Pluripotent Stem Cell (Ipsc) technology enables the generation and study of living brain tissue relevant to Parkinson's disease (PD) ex vivo. Utilizing cell lines from PD patients presents a powerful discovery system that links cellular phenotypes observed in vitro with real clinical data. Differentiating patient-derived iPSCs towards a dopaminergic (DA) neural fate revealed that these cells exhibit molecular and functional properties of DA neurons in vitro that are observed to significantly degenerate in the substantia nigra of PD patients. Clinical symptoms that drive the generation of other relevant cell types may also yield novel PD-specific phenotypes in vitro that have the potential to lead to new therapeutic avenues for patients with PD. Due to their early onset and non-familial origin, differentiated nervous tissue from these patients offer a key opportunity to discover neuron subtype-specific pathological mechanisms and importantly interrogate the contribution of their genetic background in susceptibility to PD.

Differentiation technique to generate dopaminergic neurons from induced pluripotent stem cells

Described herein are methods and compositions related to generation of induced pluripotent stem cells (iPSCs). Improved techniques for establishing highly efficient, reproducible reprogramming using non-integrating episomal plasmid vectors. Using the described reprogramming protocol, one is able to consistently reprogram non-T cells with close to 100% success from non-T cell or non-B cell sources. Further advantages include use of a defined reprogramming media E7 and using defined clinically compatible substrate recombinant human L-521. Generation of iPSCs from these blood cell sources allows for recapitulation of the entire genomic repertoire, preservation of genomic fidelity and enhanced genomic stability.

Method for reprogramming blood to induced pluripotent stem cells

The present invention teaches minimally invasive apparatuses and methods for stabilizing and/or guiding medical instruments used in a variety of medical procedures, including (a) introducing one or more substances into a subject's body, (b) removing one or more substances from a subject's body, (c) manipulating a region of a subject's body, or (d) combinations thereof. Among the many advantages of the inventive apparatuses are their simplicity and adaptability to attach to a variety of retractors.

Stabilization apparatuses and methods for medical procedures

A method of creating an isogenic multicellular blood-brain barrier model from iPSCs is disclosed.

Isogenic blood-brain barrier model

Efficient method for reprogramming blood to induced pluripotent stem cells

Growing community of inventors

Pacific Palisades, CA, United States of America

Clive N Svendsen