Methods and compositions for optimized expansion and implantation of mesenchymal stem cells
Inventors
CENTENO, CHRISTOPHER J. • KEOHAN, CRISTIN
Assignees
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Publication Number
US-9095562-B2
Publication Date
2015-08-04
Expiration Date
Abstract
Compositions and methods are provided for the optimized expansion and implantation of mesenchymal stem cells into a patient in need thereof. Autologous mesenchymal stem cells (MSCs) to a patient in need of MSCs are harvested, expanded within novel growth parameters under the influence of autologous growth factors located on the patient's platelets.
Core Innovation
The invention provides a method of expanding mesenchymal stem cells (MSCs) in vitro for use in an implant to regenerate cartilage. MSCs are obtained from an older patient in the 5th to 7th decade of life who is diagnosed with osteoarthritis, avascular necrosis, or severe degenerative joint disease. The expanded MSCs are intended to regenerate new cartilage when implanted into the patient at a site in need thereof no later than the tenth ex-vivo passage.
The method expands the MSCs with a growth medium comprising the patient’s platelet lysate in an initial amount of 10% of the growth medium for at least one passage to obtain at least 10 million to 100 million cells. The method includes determining whether the expanded MSCs are outside the scope of defined performance parameters, including doubling time of about 1 to about 3 days, cellular confluence greater than about 27 calculated according to Surface area×(% Confluence)/Cell Number, random distribution of MSCs as a monolayer culture, and greater than 30% of MSCs without a spindle shape.
When the MSCs are outside at least one parameter, the method adjusting the growth medium in subsequent passages by increasing the amount of platelet lysate to between 15-20% platelet lysate. The approach constrains ex-vivo expansion to no later than the tenth ex-vivo passage for cartilage regeneration, with dependent refinements further specifying limits and additional implantation and monitoring workflow elements.
Claims Coverage
The independent claim describes an in vitro MSC expansion method for implant-based cartilage regeneration, with multiple inventive features driven by patient-derived platelet lysate levels and defined cell performance parameters (doubling time, confluence metric, monolayer distribution, and spindle-shape constraint).
Platelet lysate-based growth medium with initial 10% and passage-dependent increase
Expanding patient-derived MSCs in vitro using a growth medium comprising the patient’s platelet lysate in an initial amount of 10% for at least one passage, and adjusting the growth medium in subsequent passages by increasing the amount of platelet lysate to between 15-20% platelet lysate when the MSCs are outside defined performance parameters.
Defined in vitro performance parameters for deciding whether to adjust growth medium
Determining whether the expanded MSCs are outside the scope of defined parameters including doubling time of about 1 to about 3 days; cellular confluence greater than about 27 calculated according to Surface area×(% Confluence)/Cell Number; random distribution of MSCs as a monolayer culture; and greater than 30% of MSCs without a spindle shape.
Cartilage regeneration after implantation by limiting ex-vivo passage
Implanting the expanded MSCs into the patient at a site in need thereof so that the expanded MSCs are capable of regenerating new cartilage no later than the tenth ex-vivo passage.
Patient selection and clinical target diagnosis
Obtaining MSCs from an older patient in the 5th to 7th decade of life diagnosed with osteoarthritis, avascular necrosis, or severe degenerative joint disease for use in implant-mediated cartilage regeneration.
Across the independent claim’s scope, inventive coverage is directed to platelet lysate-based growth media with an initial 10% level and subsequent adjustment to 15-20% based on defined in vitro performance thresholds, combined with cartilage regeneration after implantation by limiting ex-vivo passage no later than the tenth passage.
Stated Advantages
Documented Applications
Regenerating new cartilage in a patient at a site in need thereof using expanded MSCs implanted no later than the tenth ex-vivo passage.
Treatment contexts include osteoarthritis, avascular necrosis, or severe degenerative joint disease in older patients in the 5th to 7th decade of life.
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