METHOD OF CREATING HUMAN PLURIPOTENT STEM CELL DERIVED BRAIN PERICYTE-LIKE CELLS
A population of brain pericyte-like cells, wherein the cells express pericyte markers but do not express ACTA2 and wherein the cells are generated from hPSCs, is disclosed herein.
1 . A population of brain pericyte-like cells, wherein the cells express pericyte markers but do not express actin alpha 2 (ACTA2), and wherein the cells are generated from hPSCs.
2 . The population of cells of claim 1 , wherein the pericyte markers include calponin 1 (CNN1), neuron-glial antigen 2 (NG2), and platelet-derived growth factor receptor beta (PDGFRB).
3 . The population of brain pericyte-like cells of claim 1 , wherein the population is at least 90% NG2 + PDGFRB + .
4 . The cells of claim 3 , wherein the brain pericyte-like cells further express calponin and smooth muscle 22α (SM22α) but do not express alpha-smooth muscle actin (α-SMA).
5 . The cells of claim 1 , wherein the cells are capable of associating with vascular networks.
6 . The cell population of claim 1 , wherein the cells express one or more transcripts of pericyte markers selected from the group consisting of chondroitin sulfate proteoglycan 4 (CSPG4), PDGFRB, CNN1, transgelin (TAGLN), alanyl aminopeptidase (ANPEP), T-box transcription factor 18 (TBX18), ATP binding cassette subfamily C member 9 (ABCC9) and potassium inwardly rectifying channel subfamily J member 8 (KCNJ8).
7 . The cells of claim 1 , wherein the cells are capable of inducing pericyte-driven phenomena in bone marrow microvascular endothelial cells (BMECs), including the enhancement of barrier properties and reduction of transcytosis.
8 .- 12 . (canceled)
13 . A population of NCSC created by
(a) culturing hPSC in E6-CSFD medium for about 15 days to produce p75-NGFR+HNK+NCSC cells,
(b) sorting p75-NGFR + cells and re-plating the p75-NGFR + cells to produce an enriched population of p75-NGFR + NCSCs.
14 . A population of brain pericyte-like cells wherein the cells express pericyte markers but do not express ACTA2 and wherein the cells are generated from human pluripotent stem cells (hPSC), comprising the steps of
c. culturing the cells of step (b) of claim 13 in E6 media with an addition of serum for about 11 days, wherein a brain pericyte-like population of cells that express pericyte markers but do not express ACTA2 is produced.
15 . An isogenic blood-brain barrier (BBB) model created using the cells of claim 14 .
16 . A method of creating a population of p75-NGFR+HNK+NCSCs from human pluripotent stem cells, the method comprising:
a. culturing hPSC in E6-CSFD medium for about 15 days to produce p75-NGFR+HNK+NCSCs, and
b. sorting p75-NGFR + cells and re-plating the p75-NGFR + cells of step (a) to produce a population of p75-NGFR + NCSCs.
17 . The method of claim 16 , wherein step (b) is performed by magnetic activated cell sorting.
18 . The method of claim 16 , wherein the population of cells produced is p′75-NGFR + HNK + AP-2 + NCSCs which are able to double at least 5 times in culture and maintain expression p75-NGFR + , HNK + , and AP-2 + within the cells.
19 . The method of claim 16 , wherein the NCSCs are able to be maintained in culture for at least five passages and maintain NGFR + HNK + AP-2 + marker expression and do not express pericyte markers.
20 . The method of claim 16 , the method further comprising
c. culturing the cells of step (b) in E6 media with an addition of serum for about 11 days, wherein the NCSCs produce a population of a brain pericyte-like cells that express NG2, and PDGFRB but do not express ACTA2 is produced.
21 . NCSCs produced by the method of claim 16 , wherein the NCSCs maintain the potential to differentiate into neurons and mesenchymal cells.