Stem cells and pancreatic cells useful for the treatment of insulin-dependent diabetes mellitus
Inventors
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-9968639-B2
Publication Date
2018-05-15
Expiration Date
Abstract
Fresh human pancreas tissue can be used as a source of cells when to identify and select a non-stem cell population that is predisposed to be a source for surrogate pancreatic cells that can be used in treating insulin-dependent diabetes. The progenitors of these surrogate pancreatic cells have no reprogramming genes integrated into their genomes, differentiate to the pancreatic lineage pursuant to a protocol that employs only defined reagents, and are substantially unable to differentiate to the mesodermal lineage.
Core Innovation
The invention provides a composition comprising non-pluripotent progenitors of surrogate pancreatic cells, where the non-pluripotent progenitors comprise a varied population of native pancreatic cells harvested from a subject’s pancreas. These progenitors are cultured in only defined, non-animal origin reagents for fewer than five population doublings. The progenitors are unable to differentiate into the mesodermal lineage, but can differentiate into surrogate pancreatic cells suitable for treating insulin-dependent diabetes.
The invention further defines surrogate pancreatic cells produced from the non-pluripotent progenitors by their ability to survive and differentiate using a differentiation protocol that comprises defined, non-animal-origin components. The protocol is directed toward producing a large population of pancreatic cells efficiently, while maintaining the inability of the non-pluripotent progenitors to differentiate into the mesodermal lineage. In some embodiments, the differentiation protocol includes lineage progression to an endodermal lineage followed by pancreatic endocrine lineage differentiation.
Surrogate pancreatic cell compositions are characterized by pancreatic endocrine marker expression and by restrictions on genetic modification and protocol components. In particular, non-pluripotent progenitors may have no reprogramming genes integrated into their genomes, and produced pancreatic cells may be defined by high expression of Pdx1, Nkx6.1, and NeuroD1, including threshold values such as at least about 80% or at least about 90% expressing genes characteristic of pancreatic endocrine progenitors, and more than about 90% expressing the markers Pdx1, Nkx6.1, and NeuroD1. The differentiation protocol can further exclude contacting the progenitors with a wnt family member.
Claims Coverage
The partial content includes two independent claims. Across these, the inventive features focus on non-pluripotent progenitors derived from native pancreatic cells with defined, non-animal origin culture limitations and mesodermal lineage exclusion, and on survivability and efficient production of pancreatic cells using a defined, non-animal-origin differentiation protocol that maintains mesodermal lineage exclusion while enabling pancreatic endocrine differentiation, with defined marker and optional genetic/protocol constraints.
Non-pluripotent surrogate pancreatic progenitors from native pancreas in defined non-animal origin reagents for fewer than five population doublings
The progenitors comprise a varied population of native pancreatic cells harvested from a subject’s pancreas, cultured in only defined, non-animal origin reagents for fewer than five population doublings.
Mesodermal lineage exclusion while differentiating into surrogate pancreatic cells for insulin-dependent diabetes
The non-pluripotent progenitors are unable to differentiate into the mesodermal lineage, but can differentiate into surrogate pancreatic cells that are suitable for treating insulin-dependent diabetes.
Survival through defined non-animal-origin differentiation protocol to efficiently produce a large population of pancreatic cells
The non-pluripotent progenitors have the ability to survive a differentiation protocol such that a large population of pancreatic cells can be efficiently produced, where the differentiation protocol comprises defined, non-animal-origin components.
Optional two-stage lineage differentiation to endodermal then pancreatic endocrine lineage
In the differentiation protocol, a first combination of components drives cells to an endodermal lineage, and endodermal cells are then exposed to a second combination to drive pancreatic endocrine lineage differentiation.
High pancreatic endocrine marker expression defining differentiated surrogate pancreatic cells
Cells produced can be characterized by high expression of genes characteristic of pancreatic endocrine progenitors and/or by marker expression thresholds, including at least about 80% or at least about 90% expressing genes characteristic of pancreatic endocrine progenitors, and more than about 90% expressing Pdx1, Nkx6.1, and NeuroD1.
No reprogramming genes integrated into progenitor genomes
The progenitors have no reprogramming genes integrated into their genomes.
Exclusion of contacting progenitors with a wnt family member
The differentiation protocol does not comprise contacting the progenitors with a wnt family member.
Taken together, the independent claims define compositions of non-pluripotent surrogate pancreatic progenitors derived from native pancreatic cells that cannot differentiate into the mesodermal lineage, but can generate surrogate pancreatic cells suitable for treating insulin-dependent diabetes. Differentiation is supported by defined, non-animal-origin components, with optional two-stage endodermal-to-pancreatic endocrine progression, optional exclusion of wnt family member contact, optional absence of integrated reprogramming genes, and marker-based definitions using Pdx1, Nkx6.1, and NeuroD1 with quantitative thresholds.
Stated Advantages
Surrogate pancreatic cells suitable for treating insulin-dependent diabetes can be produced from non-pluripotent progenitors.
The non-pluripotent progenitors cultured in defined, non-animal origin reagents for fewer than five population doublings can be used to generate surrogate pancreatic cells.
The differentiation protocol allows the non-pluripotent progenitors to survive while efficiently producing a large population of pancreatic cells.
Produced pancreatic cell populations can be defined by high expression of pancreatic endocrine markers (Pdx1, Nkx6.1, and NeuroD1) and/or high proportions of cells expressing genes characteristic of pancreatic endocrine progenitors.
Documented Applications
Treating insulin-dependent diabetes using surrogate pancreatic cells differentiated from non-pluripotent progenitors of surrogate pancreatic cells.
Interested in licensing this patent?