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US 12,110,510 B2Granted
Functional cortico-spinal-muscle assembled spheroids
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsJimena Andersen, Sergiu P. Pasca
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Abstract
Functional human cortico-spinal-muscle assembled spheroids are generated by in vitro culture. Complete cortico-spinal-muscle spheroids (hCS-hSC-hSkM) are assembled from component cultured cell systems, where each cultured cell system is designed to provide specific sets of neural and/or muscle cells, and which components are functionally integrated in the assembled spheroid.
Claims17
A method for producing functionally integrated human cortico-spinal-muscle assembled spheroids in vitro, the method comprising:
- (i) inducing in a human pluripotent stem cell suspension culture a neural fate to provide a first spheroid of neural progenitor cells and a second spheroid of neural progenitor cells;
- (ii) differentiating the neural progenitor cells in the first spheroid to differentiate into human spinal cord spheroids (hSC), and culturing the hSC with human skeletal muscle cells (hSkM) under integrating conditions to generate hSC-hSkM spheroids;
- (iii) differentiating the neural progenitor cells in the second spheroid to differentiate into human cortical spheroids (hCS);
- (iv) culturing the hCS and hSC-hSkM under conditions permissive for spheroid fusion while maintaining for an extended period of time in neural medium;
- wherein a functionally integrated human cortico-spinal-muscle assembled spheroid is differentiated comprising interacting glutamatergic projection neurons, motor neurons, interneurons, and muscle cells.
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Dependencies are inferred from the claim text.