Improved Cortical Spheroids and Methods of Making the Same

Abstract

Improved hybrid neurovascular spheroids and methods for making the same are provided. In some embodiments of a method for making a hybrid neurovascular spheroid, the method includes i) propagating cortical cells to form a cortical spheroid; ii) propagating endothelial cells to form an endothelial spheroid; iii) propagating mesenchymal stem cells to form a mesenchymal cell culture; and iv) combining the cortical spheroid, endothelial spheroid, and mesenchymal spheroid under conditions to form the hybrid neurovascular spheroid.

That which is claimed is: 
     
         1 . A hybrid neurovascular spheroid comprising cortical cells, endothelial cells, and mesenchymal stem cells. 
     
     
         2 . The hybrid neurovascular spheroid of  claim 1 , wherein the hybrid neurovascular spheroid displays upregulated secretion levels of one or more cytokines VEGF-A, PGE2, and TBF-β1 relative to a comparative hybrid neurovascular spheroid prepared by directly mixing cultures of cortical cells, endothelial cells, and mesenchymal stem cells. 
     
     
         3 . The hybrid neurovascular spheroid of  claim 1 , wherein the hybrid neurovascular spheroid displays increased expression of one or more of TBR1, Nkx2.1, matrix remodeling, MMP2, MMP3, Notch-1, GLUT-1, CD31, or ZO-1 genes relative to a comparative hybrid neurovascular spheroid prepared by directly mixing cultures of cortical cells, endothelial cells, and mesenchymal stem cells. 
     
     
         4 . The hybrid neurovascular spheroid of  claim 1 , wherein the cortical cells are human iPSC-derived cortical neural progenitor cells. 
     
     
         5 . The hybrid neurovascular spheroid of  claim 1 , wherein the endothelial cells are human endothelial cells. 
     
     
         6 . The hybrid neurovascular spheroid of  claim 1 , wherein the mesenchymal stem cells are human mensenchymal stem cells (hMSC). 
     
     
         7 . The hybrid neurovascular spheroid of  claim 1 , wherein the cortical cells are human induced pluripotent stem cells (iPSC)-derived cortical neural progenitor cells, and wherein the endothelial cells are human endothelial cells. 
     
     
         8 . The hybrid neurovascular spheroid of  claim 1 , wherein the cortical cells are human iPSC-derived cortical neural progenitor cells, and wherein the mesenchymal stem cells are human mensenchymal stem cells (hMSC). 
     
     
         9 . The hybrid neurovascular spheroid of  claim 1 , wherein the cortical cells are human iPSC-derived cortical neural progenitor cells, wherein the endothelial cells are human endothelial cells, and wherein the mesenchymal stem cells are human mensenchymal stem cells (hMSC). 
     
     
         10 . The hybrid neurovascular spheroid of  claim 1 , wherein a ratio based on initial seeded cell numbers of the cortical cells to the endothelial cells to the mesenchymal stem cells is from about 4:2:0 to about 1:2:3. 
     
     
         11 . The hybrid neurovascular spheroid of  claim 1 , wherein one or more of the cortical cells, endothelial cells, and mesenchymal stem cells are propagated in a hydrogel comprising basement membrane, hyaluronic acid, or any combination thereof. 
     
     
         12 . The hybrid neurovascular spheroid of  claim 1 , wherein the hybrid neurovascular spheroid is an antagonist-treated hybrid neurovascular spheroid. 
     
     
         13 . A hybrid neurovascular spheroid comprising cortical cells, endothelial cells, and mesenchymal stem cells, wherein the hybrid neurovascular spheroid displays upregulated secretion levels of one or more cytokines VEGF-A, PGE2, and TBF-β1 relative to a comparative hybrid neurovascular spheroid prepared by directly mixing cultures of cortical cells, endothelial cells, and mesenchymal stem cells.