IP Library › Granted Patent US 12,441,987
Granted Patent B2
US 12,441,987 · App. 17/458,838 · Granted Oct 14, 2025

Multicellular integrated brain tissue in neurological diseases

Inventors: Li-Huei Tsai (Cambridge, MA); Joel Blanchard (Arlington, MA); Alice Stanton (Boston, MA); Adele Bubnys (Cambridge, MA); Robert S. Langer (Newton, MA)
Assignee: Massachusetts Institute of Technology
C12N5/0691A61K35/30C12N5/0062C12N5/0607C12N5/0622C12N2502/28C12N2503/00C12N2513/00C12N2533/90
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Quick Facts
Patent No.
US 12,441,987
App. No.
17/458,838
Granted
Oct 14, 2025
Kind
B2
Abstract

The present disclosure provides, in some embodiments, in vitro brain (miBRAIN) having functional and structural properties of in vivo brain as well as methods of identifying compounds capable of influencing brain function.

Claims (20)

1. A multi-cellular integrated micro brain (miBRAIN) comprising a 3-dimensional (3D) matrix comprising an interconnected cerebrovascular network comprised of human brain endothelial cell (BEC) vessels positioned in the 3D matrix and comprised of human pluripotent stem cell-derived endothelial cells, human pluripotent stem cell-derived pericytes proximal to the BEC vessel and physically interacting with an apical surface of the BEC vessels, human pluripotent stem cell-derived astrocytes, human myelinated cells, each of which is co-cultured in and dispersed throughout the 3D matrix, and optionally immune cells, wherein the 3D matrix comprises a material selected from the group consisting of an engineered hydrogel, exogenous RGD polymers and dextran.

2. A multi-cellular integrated micro brain (miBRAIN) comprising a 3-dimensional (3D) matrix comprising an interconnected cerebrovascular network comprised of human pluripotent stem cell-derived endothelial cells, human pluripotent stem cell-derived pericytes, human pluripotent stem cell-derived astrocytes, human pluripotent stem cell-derived neuron cells, human pluripotent stem cell-derived microglial cells, and human oligodendrocytes or oligodendrocyte precursor cells (OPCs), each of which is co-cultured in, housed within, and dispersed throughout the 3D matrix, and forms the interconnected cerebrovascular network, wherein the 3D matrix comprises a material selected from the group consisting of an engineered hydrogel, exogenous RGD polymers and dextran.

3. The miBRAIN of claim 1 , wherein the 3D matrix comprises an extracellular matrix (ECM) substrate.

4. The miBRAIN of claim 1 , wherein the 3D matrix comprises type-I collagen.

5. The miBRAIN of claim 1 , wherein the 3D matrix comprises a solubilized basement membrane preparation.

6. The miBRAIN of claim 1 , wherein the 3D matrix comprises arginine, glycine, aspartic acid (RGD)-polymer.

7. The miBRAIN of claim 1 , wherein the microglial cells are derived from human hematopoietic progenitor cells (HPCs).

8. The miBRAIN of claim 1 , wherein the astrocytes in miBRAIN exhibit radial morphologies with numerous processes extending from the cell body.

9. The miBRAIN of claim 1 , wherein the endothelial cells express endothelial cell markers platelet endothelial cell adhesion molecule (PECAM) and/or vascular endothelial-cadherin (VE-CAD).

10. The miBRAIN of claim 1 , wherein the pericytes express neuron-glial antigen 2 (NG2) and/or platelet derived growth factor receptor beta (PDGFrB).

11. The miBRAIN of claim 1 , further comprising a culture media comprising human endothelial serum-free medium (hESFM), astrocyte growth supplement (AGS), and B27.

12. The miBRAIN of claim 11 , wherein the culture media comprises human endothelial serum-free medium (hESFM), astrocyte growth supplement (AGS), antioxidants, catalase, superoxide dismutase and one or more supplements selected from the group consisting of neurotrophin-3 (NT3), triiodothyronine (T3), insulin-like growth factor (IGF), platelet derived growth factor subunit B (PDGFB), cyclic adenosine monophosphate (cAMP), amino acids (AA), brain-derived neurotrophic factor (BDNF), Glial cell derived neurotrophic factor (GDNF), macrophage colony-stimulating factor (mCSF), interleukin 34 (IL34), and vascular endothelial growth factor A (VegF-A).

13. The miBRAIN of claim 1 , further comprising at least one biochemical factor selected from the group consisting of: RGD peptide, collagen, fibronectin, laminin, aggrecan, hyaluronic acid, heparin, reelin, tenascin-R (TenR), tenascin-C (TenC), brevican (BCN), versican (VCN), neurocan (NCN), and thrombospondin-1 (TSP1).

14. The miBRAIN of claim 13 , wherein the at least one biochemical factor comprises an RGD peptide.

15. The miBRAIN of claim 1 , wherein at least one cell is an iPSC-derived cell.

16. The miBRAIN of claim 1 , wherein at least one cell is an APOE4 neuron.

17. The miBRAIN of claim 1 , wherein the astrocytes express GFAP and S100b.

18. The miBRAIN of claim 1 , wherein the 3D matrix is comprised of dextran.

19. The miBRAIN of claim 1 , further comprising human pluripotent stem cell-derived neurons.

20. The miBRAIN of claim 1 , further comprising human pluripotent stem cell-derived microglial cells.

Assignments (2)
CONFIRMATORY LICENSE Recorded Dec 13, 2023
From: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 065988/0597 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 26, 2022
From: TSAI, LI-HUEI; BLANCHARD, JOEL; STANTON, ALICE; BUBNYS, ADELE; LANGER, ROBERT S.
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 060202/0079 →
Continuity (2)
Provisional Application 63106320 · Oct 27, 2020
Related Publication 20220127581A1 · Apr 28, 2022
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