Stem cells within gel microenvironments
First Claim
1. A method of encapsulating stem cells within three-dimensional polymer microenvironments, wherein said polymer is a naturally derived polymer obtained from living organisms, wherein said method comprises the following steps:
- (i) mixing a polymer solution with stem cells in a buffer to incorporate and disperse the stem cells therein to form a cell-liquid suspension, (ii) emulsifying said cell-liquid suspension in a hydrophobic fluid phase to form an emulsified liquid, (iii) initiating polymerization of said emulsified liquid to form three-dimensional polymer microenvironments, and (iv) collecting the three-dimensional polymer microenvironments from the hydrophobic phase.
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Abstract
This invention provides a system to embed stem cells within three-dimensional (3D) hydrogel microenvironments consisting of naturally derived proteins, proteoglycans and/or polysaccharides. Pure matrices or combinations of materials can be used. The method involves suspending stem cells in solutions of the matrix components of interest, emulsifying these solutions in a hydrophobic phase, triggering gelation of the matrix components by changing the environmental conditions, and collection of the resulting hydrogel beads. The unique bead format of this invention has the advantage of allowing the use of small amounts of rare matrix proteins. Bead preparations can be concentrated into a paste for use as a cell delivery vehicle to damaged tissues, either directly after encapsulation or after a period of culture to promote stem cell differentiation. Defined 3D microenvironments can guide stem cell differentiation, and the resulting beads can be used directly as a cell delivery vehicle in various tissue repair applications.
77 Citations
45 Claims
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1. A method of encapsulating stem cells within three-dimensional polymer microenvironments, wherein said polymer is a naturally derived polymer obtained from living organisms, wherein said method comprises the following steps:
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(i) mixing a polymer solution with stem cells in a buffer to incorporate and disperse the stem cells therein to form a cell-liquid suspension, (ii) emulsifying said cell-liquid suspension in a hydrophobic fluid phase to form an emulsified liquid, (iii) initiating polymerization of said emulsified liquid to form three-dimensional polymer microenvironments, and (iv) collecting the three-dimensional polymer microenvironments from the hydrophobic phase. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A method of producing a population of differentiated viable stem cells, wherein said method comprises directed differentiation of stem cells by the following steps:
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(i) mixing a polymer solution derived from living organisms with stem cells in a buffer to incorporate and disperse the stem cells therein to form a cell-liquid suspension, (ii) emulsifying said cell-liquid suspension in a hydrophobic fluid phase to form an emulsified liquid, (iii) initiating polymerization of said emulsified liquid to form three-dimensional polymer microenvironments, (iv) collecting the three-dimensional polymer microenvironments via centrifugation, and (v) culturing said three-dimensional polymer microenvironments to promote the directed differentiation of the embedded stem cells. - View Dependent Claims (19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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31. A method of encapsulating and delivering stem cells within hydrogel polymer microenvironments, wherein said method comprises the following sequential steps:
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(i) mixing a polymer derived from living organisms with stem cells in a buffer to incorporate and disperse the stem cells therein to form a cell-liquid suspension, (ii) emulsifying said cell-liquid suspension in a hydrophobic fluid phase to form an emulsified liquid, (iii) initiating polymerization of said emulsified liquid to form polymer microenvironments, (iv) collecting the polymer microenvironments from the hydrophobic phase, (v) forming a concentrated paste of said polymer microenvironments, and (vi) loading the paste into a syringe and extruding it for cosmetic or therapeutic application to a subject in need thereof. - View Dependent Claims (32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45)
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Specification