Medium, methods, cells and secreted factors for stem cell culture and therapy

Inventors

Betancourt, Aline M. • Waterman, Ruth S. • ISETT, Thomas F.

Assignees

Sanbio Inc

Abstract

Described herein are methods and culture medium, useful for inducing polarization in multipotent stem cells. Additionally, described herein are multipotent cells produced by the methods and culture medium of this disclosure that are useful therapeutic agents. Also described herein are extracellular vesicles and factors secreted by multipotent cells that are produced by the methods and culture medium of this disclosure that are useful as therapeutic agents.

Core Innovation

The invention relates to producing an immunologically polarized mesenchymal stem cell population from an unstimulated mesenchymal stem cell population. It is achieved by contacting the unstimulated mesenchymal stem cell population with a culture medium comprising a Toll-like receptor 3 (TLR3) ligand and an agent selected from acetylcholine, norepinephrine, epinephrine, serotonin, and melatonin, and expression of CXCL9 is induced.

The disclosed polarization approach uses Toll-like receptor signaling combined with a second inducing agent to generate discrete type 1 (pro-inflammatory) or type 2 (anti-inflammatory) phenotypes. Type 2 polarization is associated with induction of CXCL9 and corresponding miRNA signatures, while type 1 polarization is associated with induction of TNFSF10 (TRAIL) and corresponding miRNA signatures.

The disclosure further characterizes secreted by-products from the polarized mesenchymal stem cells, including conditioned factors and extracellular vesicles (EVs) with altered protein content and miRNA signatures. It associates elevated or reduced EV protein markers with the pro-inflammatory versus anti-inflammatory polarized phenotypes.

Claims Coverage

The provided content includes one independent claim directed to producing an immunologically polarized mesenchymal stem cell population using a TLR3 ligand plus a selected agent, with CXCL9 induction as the stated outcome.

The core inventive elements are the use of a TLR3 ligand together with a selected agent from the neurotransmitter/melatonin group to immunologically polarize mesenchymal stem cells, with CXCL9 expression as the induced marker.

Stated Advantages

Documented Applications