Carolina Machado Email & Phone Number

London, Gb

London, Gb

Cambridge, United Kingdom

Target validation and pathway discovery employing human stem cell models of Alzheimer’s disease.Drug screening - High Content Imaging - Project Management

Assembly of neuromuscular circuits from stem cell-derived components in vitro.My study focus on the formation of neuromuscular circuits in vitro to model degenerative diseases. To this end, I am assembling the cellular components of the system, motor neurons, glia and muscle, in compartmentalized tissue culture devices. The derivation of these cell types from stem cells relies on understanding the development of different types of tissue and organs in vivo. I have started this project employing murine embryonic stem cells but I am now transferring the knowledge gained from the murine system towards making human models of degenerative disease. I have genetically modified murine and human stem cells to allow the sorting of specific cell types (neurons, glia and myoblasts) from mixed cultures. I am assembling these cell types in tissue culture devices divided into compartments, which allows the segregation of CNS and periphery, further improving the in vivo-likeness of the system. This approach will be applied to study Amyotrophic Lateral Sclerosis (ALS), a fatal neurodegenerative disease without effective treatment.Expertise employed in this project:- human pluripotent stem cell culture- mouse embryonic stem cell culture- generation of transgenic cell lines using CRISPR/Cas9, TALENs, PiggyBac, Tol2- High content imaging- Microfluidics and compartmentalised devices- Flow cytometry and magnetic activated cell sorting- Differentiation models, neural and myoblast- Organoids and 3D co-cultures- Optogenetics- Long term culture of neurons, glia and muscle.- qPCR- Confocal Imaging

The molecular neurobiology of breathing.My work has focused in the acquisition of phrenic motor neuron fate in the spinal cord. Phrenic or respiratory neuron control respiratory muscles relaying the breathing rhythm generated by the brainstem. The transcriptional program of phrenic neuron identity was investigated using a combination of in vitro differentiation of motor neurons from mouse ES cells, mouse genetics, imaging and systematic analysis of gene expression. This study provided a framework for understanding how phrenic neuron identity is conferred in vivo using stem cells as a tool to dissect key events in the development of neural tissue.Expertise employed in this project:- mouse embryonic stem cell culture- generation of transgenic cell lines- flow cytometry and magnetic activated cell sorting- differentiation models, motor neurons- cloning- gene expression profiling- confocal Imaging- imaging analysis (ImageJ)- whole embryo culture- embryo dissection and manipulation

United Kingdom

www.nibsc.orgDivision of Cell Biology and Imaging - NIBSCThe project developed a general technology for quickly and efficiently examining large numbers of linked variables, addressing a central issue of regenerative medicine; the capacity to reproducibly control the differentiation of a high proportion of stem cells. This project was part of a consortium bringing together strengths in cell biology, advanced imaging, robotic engineering and combinatorial techniques.Expertise employed in this project:- Human embryonic stem cell culture- Neural differentiation - Imaging analysis - Confocal Imaging- Cryo-SEM- Microcarrier culture based assays- Live cell imaging- Ethical and regulatory aspects involving human embryonic stem cell research- Stem cell banking- Product development