Pluripotent Stem Cells Differentiate into Motor Neurons in Cynomolgus Monkeys

MN (motor neuron) is a specialized type of neuron whose cell body is located in the motor cortex, brainstem, and the spinal cord. The damage of this type of cell results in many neurodegenerative diseases. So far, it is extremely difficult to study these diseases in vivo. Therefore, it is of great significance to study the pathogenesis and therapy of neurodegenerative diseases through the differentiation of mature MN in vitro. Cynomolgus monkeys (Macaca fascicularis), as a non-human primate model, has a highly similar genetic background with humans and is widely used for the study of these diseases. We developed a method to guide the induced pluripotent stem cell of cynomolgus monkeys into an enriched population (>90%) of mature MNs in 28 days, which consists of four differentiation stages. The four stages are as follows: (1) iPSCs (induced pluripotent stem cells) were induced into NEP (neuroepithelial progenitors); (2) NEP were induced into MNP (motor neuron progenitors); (3) MNP were induced into non-mature MN; (4) non-mature MN were induced into mature MN. This study successfully established a method for guiding iPSCs of cynomolgus monkeys into MN. Moreover, it showed that the poly-L-ornithine and lamina-521 double-coated dish, cell density, and the addition of ROCK inhibitor (Y27632) can contribute to the survival of MN. In summary, this study provides a basis for studying the pathogenesis and stem cell transplantation therapy of MN degenerative diseases.

CSTR: 32200.14.cjcb.2019.10.0015