Wnt-activated adipose-derived stem cell apparatuses, methods and systems
AssigneeRegeneration Biomedical, Inc.
InventorChristopher Duma
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Abstract
The WNT-ACTIVATED ADIPOSE-DERIVED STEM CELL APPARATUSES, METHODS AND SYSTEMS (hereinafter “WAADSC”) disclosed herein in various embodiments provide for production of an isolated and enriched population of mesenchymal stem cells that have an active Wnt signaling demonstrated by the elevated expression of Lgr5 marker and/or Nestin in more than 50% of the population. Such an autologous cell population may, in embodiments, be injected into cerebral ventricles of patients with neurodegenerative diseases to yield therapeutic results, such as halting the progression of certain conditions and/or ameliorating specific symptoms thereof.
Claims20
A method for producing adipose-derived Wnt-activated mesenchymal stem cells, comprising:
- suspending adipose mesenchymal stem cells, blood progenitors, and a mixture of fibroblasts, myocytes, vascular endothelial cells, and blood cells in a culture flask with media with antibiotic and a dissociating enzyme to yield a first suspension;
- incubating the first suspension;
- centrifuging the first suspension;
- removing a first supernatant;
- adding the media, signaling amino acids, and growth factors to yield a second suspension,
- wherein the growth factors comprise at least one of Activin A and basic Fibroblast Growth Factor, and
- wherein the signaling amino acids comprise at least one of leucine and taurine but not arginine;
- incubating the second suspension;
- removing a second supernatant;
- adding the media and the growth factors to yield a third suspension;
- applying at least one recombinant cell-dissociation enzyme to the third suspension;
- centrifuging the third suspension;
- removing a third supernatant;
- adding the media and the growth factors to yield a fourth suspension;
- homogenizing the fourth suspension;
- and
- adding the media and the growth factors to yield confluent adipose-derived Wnt-activated mesenchymal stem cells,
- wherein more than 50% of the adipose-derived Wnt-activated mesenchymal stem cells exhibit elevated expression of Lgr5 and/or Nestin compared to isolated adipose-derived mesenchymal stem cells not subjected to the method for producing adipose-derived Wnt-activated mesenchymal stem cells.
Dependencies are inferred from the claim text.