Compositions and methods for using small mobile stem cells
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Publication Number
US-12146164-B2
Publication Date
2024-11-19
Expiration Date
Abstract
Disclosed herein are small mobile stem (SMS) cells and methods of culturing, isolating, and using SMS cells. Also disclosed are methods of culturing SMS cells in an undifferentiated state for prolonged periods of time, and for using SMS cells for the production of a variety of molecules, including proteins, extracellular matrix (ECM) proteins, and the use of SMS cells in microfluidic devices.
Core Innovation
The disclosure relates to maintaining an isolated population of small mobile stem (SMS) cells in suspension culture in an undifferentiated state for a prolonged period. The SMS cells are cultured in a polypropylene vessel or a pretreated siliconized vessel containing high glucose Dulbecco’s Modified Eagle Medium (DMEM) comprising calf serum, configured to promote growth of the SMS cells in suspension.
The disclosure further characterizes the SMS cells as adherent equi-dimensional cells with strict radial symmetry, a translucent cytoplasm, and a diameter of 4.5 to 5.5 µm, with high mobility. The SMS cells are derived from umbilical cord, peripheral blood, bone marrow, or solid tissue, and are handled as a suspension population, with embodiments describing separation of suspended SMS cells from adherent cells.
The disclosure also describes isolating or purifying undifferentiated SMS cells by differential centrifugation or filtration in order to obtain an undifferentiated SMS cell population. The disclosed system includes optional microfluidic systems and further processing embodiments for downstream use, including producing molecules, including extracellular matrix (ECM) proteins derived from SMS cells, with optional transfection and induction to produce gene products.
Claims Coverage
The independent claim covers an SMS suspension culturing method that maintains an isolated population of SMS cells in suspension in an undifferentiated state using a polypropylene or pretreated siliconized vessel with high glucose DMEM comprising calf serum. The claim further specifies physical and morphological characteristics of the SMS cells and the source tissues. Dependent claim coverage includes vessel formats, optional surface etching, microfluidic channel embodiments, medium movement/circulation, and separating suspended SMS cells from adherent cells.
Suspension culturing SMS cells in polypropylene or siliconized vessel to maintain undifferentiated state
Culturing an isolated population of small mobile stem (SMS) cells in a vessel configured to promote growth of the SMS cells in a suspension, wherein the vessel is a polypropylene vessel or a pretreated siliconized vessel containing high glucose Dulbecco’s Modified Eagle Medium (DMEM) comprising calf serum, thereby maintaining the isolated population of SMS cells in suspension in an undifferentiated state.
SMS cell morphology and mobility in suspension
Using SMS cells that are adherent equi-dimensional cells with strict radial symmetry, exhibit a translucent cytoplasm, are from 4.5 to 5.5 µm in diameter, and are highly mobile.
SMS cell source from multiple tissues
Using SMS cells derived from umbilical cord, peripheral blood, bone marrow, or solid tissue.
Medium movement or circulation during suspension culture
Further culturing the cells with movement or circulation of the medium.
Suspension culturing in selectable vessel formats
Performing the method in a vessel selected from a flask, container, chamber, channel, tube, niche, or bioreactor.
Etched surface of vessel component
Further including etching at least part of a surface of a flask or other specified container or bioreactor component.
Microfluidic channel embodiment
Wherein the channel used is a microfluidic channel.
Separating suspended SMS cells from adherent cells
Further including separating an isolated population of suspended SMS cells from adherent cells.
Overall, the claims focus on suspension culturing SMS cells in specific vessel types and media to maintain undifferentiated suspension conditions, together with defined cell morphology, mobility, and specified tissue sources. Dependent claim refinements expand vessel embodiments, add optional vessel surface etching and medium movement/circulation, and include separating suspended SMS cells from adherent cells.
Stated Advantages
Documented Applications
Producing molecules, including proteins and SMS-derived extracellular matrix (ECM) proteins, optionally using transfection and induction with chemical inducers.
Differentiating SMS cells into multiple lineages including neural, osteogenic, and adipogenic lineages.
Generating tissue-like structures and scaffolded extracellular matrix and using these for wound healing, including wound-healing systems and wound-dressing materials involving SMS-derived antimicrobial ECM proteins.
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