Altered adherent stromal cells and methods of producing and using same
Inventors
Abraham, Eytan • Raviv, Lior • WADAMANY, Maya • ESHKOL, Nadav
Assignees
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Publication Number
US-10851347-B2
Publication Date
2020-12-01
Expiration Date
Abstract
Disclosed herein are altered adherent stromal cells and methods of producing and utilizing same.
Core Innovation
The disclosure relates to altered and modified adherent stromal cells (ASC) derived from placenta and/or adipose tissue, including ASC generated in a three-dimensional (3D) culture. ASC are expanded after a two-dimensional (2D) culture step and then transferred to a 3D culture format in a bioreactor with a cytokine-containing growth medium in which one or more cytokines are added, and cytokine incubation is commenced during exponential phase.
Cytokine-treated ASC exhibit distinct properties, including elevated therapeutic factor secretion and immunomodulatory effects. Cytokine stimulation is associated with increased secretion of factors such as VEGF, IL-6, MCP-1, GM-CSF, RANTES, and CCL5, and the disclosure also characterizes manufacturing and culture formats used to obtain the ASC.
The invention also provides modified ASC populations defined by multi-marker expression patterns at the population level, including combinations of CD10, CD29, CD38, CD40, HLA-DR, and CD74. The described modified ASC populations include further defined variants characterized by additional expression markers such as CD90, quantified marker positivity, and absence of certain markers, and are further distinguished by reduced differentiation into osteocytes and/or adipocytes under conditions that drive classical MSC differentiation.
Claims Coverage
The independent claims cover three inventive features. One covers increasing CCL5 secretion from ASC using a cytokine-containing 3D bioreactor culture commenced during exponential phase, and two cover characterized cell populations defined by majority expression of specified marker combinations at the population level.
Incubating placental or adipose ASC in a cytokine-containing 3D bioreactor culture with TNF-alpha started during exponential phase to increase CCL5 secretion
A method of increasing CCL5 secretion from ASC by incubating ASC from placental or adipose tissue in a 3D culture in a cytokine-containing growth medium comprising TNF-alpha, where the 3D culture is performed in a bioreactor, and where incubating in the cytokine-containing growth medium is commenced during exponential phase.
Cell population majority expresses CD10, CD29, CD38, and CD40 at the population level
A population of cells in which the majority of the population expresses on a population level each of CD10, CD29, CD38, and CD40.
Cell population majority expresses CD10, CD29, HLA-DR, and CD74 at the population level
A population of cells in which the majority of the population expresses on a population level each of CD10, CD29, HLA-DR, and CD74.
Across the independent claims, the disclosure covers increasing CCL5 secretion from placental or adipose ASC via cytokine-containing 3D bioreactor culture with TNF-alpha and exponential-phase commencement, and defined ASC cell populations characterized by majority expression of specified marker combinations at the population level.
Stated Advantages
Increasing CCL5 secretion from ASC.
elevated therapeutic factor secretion associated with cytokine-conditioned ASC.
immunomodulatory effects associated with cytokine-conditioned ASC.
Reduced differentiation into osteocytes and/or adipocytes under conditions that drive classical MSC differentiation.
Supports therapeutic use across immune-mediated diseases, transplant rejection, ischemia including critical limb ischemia, myocardial infarction, angiogenesis, hematopoietic stem cell (HSC) engraftment, and fibrosis including pulmonary fibrosis.
Provides secreted extracellular vesicles (exosomes) and conditioned media (CM) derived from the modified ASC populations for downstream therapeutic activity.
Documented Applications
Generation of altered adherent stromal cells (ASC) from placenta and/or adipose tissue in 3D bioreactor culture with cytokine stimulation.
Characterized cell populations used as pharmaceutical compositions.
Characterized cell populations included in a bioreactor.
Immune-mediated diseases.
Transplant rejection.
Ischemia, including critical limb ischemia.
Myocardial infarction.
Angiogenesis.
Hematopoietic stem cells (HSC) engraftment.
Fibrosis, including pulmonary fibrosis.
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