Encapsulation and cardiac differentiation of hiPSCs in 3D PEG-fibrinogen hydrogels
The present invention relates to the production of cell cultures and tissues from undifferentiated pluripotent stem cells using three-dimensional biomimetic materials. The resultant cell cultures or tissues can be used in any of a number of protocols including testing chemicals, compounds, and drugs. Further, the methods and compositions of the present invention further provide viable cell sources and novel cell delivery platforms that allow for replacement of diseased tissue and engraftment of new cardiomyocytes from a readily available in vitro source. The present invention includes novel methods required for the successful production of cell cultures and tissues, systems and components used for the same, and methods of using the resultant cell and tissue compositions.
1. A three-dimensional, synchronously contracting cardiac tissue comprising;
a hydrogel material comprising a covalently crosslinkable component and a natural hydrogel component, the natural hydrogel component comprising one or more of fibrinogen, collagen, gelatin, hyaluronic acid, elastin, fibronectin, laminin, fibrin, alginate, and decellularized cardiac extracellular matrix, and
three-dimensionally differentiated, PSC-derived cardiomyocytes encapsulated within the hydrogel material,
wherein the three-dimensionally differentiated, PSC-derived cardiomyocytes have been differentiated from PSCs within the hydrogel material.
2. The three-dimensional, synchronously contracting cardiac tissue of claim 1 , wherein the synchronously contracting cardiac tissue contracts as a single unit.
3. The three-dimensional, synchronously contracting cardiac tissue of claim 2 , wherein the synchronously contracting cardiac tissue contracts spontaneously.
4. The three-dimensional, synchronously contracting cardiac tissue of claim 3 , wherein the spontaneous contraction frequency ranges from 0.59 to 1.53 Hertz.
5. The three-dimensional, synchronously contracting cardiac tissue of claim 1 , wherein the three-dimensionally differentiated, PSC-derived cardiomyocytes are in situ-differentiated cardiomyocytes.
6. The three-dimensional, synchronously contracting cardiac tissue of claim 1 , wherein the PSC-derived cardiomyocytes are axially aligned to each other.
7. The three-dimensional, synchronously contracting cardiac tissue of claim 6 , wherein the PSC-derived cardiomyocytes comprise Z-bands aligned with T-tubules.
8. The three-dimensional, synchronously contracting cardiac tissue of claim 6 , wherein the PSC-derived cardiomyocytes comprise sarcomeres spaced by 1.8 micrometers to 2.0 micrometers.
9. The three-dimensional, synchronously contracting cardiac tissue of claim 6 , wherein the PSC-derived cardiomyocytes comprise H-bands.
10. The three-dimensional, synchronously contracting cardiac tissue of claim 6 , wherein the PSC-derived cardiomyocytes comprise basement membrane with caveolae.
11. The three-dimensional, synchronously contracting cardiac tissue of claim 6 , wherein polar ends of the PSC-derived cardiomyocytes comprise intercalated discs and gap junctions.
12. The three-dimensional, synchronously contracting cardiac tissue of claim 1 , wherein the three-dimensional, synchronously contracting cardiac tissue is shaped as a microisland, a cardiac disc, a cardiac string, a macrotissue, or a microsphere.
13. The three-dimensional, synchronously contracting cardiac tissue of claim 1 , wherein the covalently crosslinkable component comprises an acrylate component.
14. The three-dimensional, synchronously contracting cardiac tissue of claim 1 , wherein the PSC-derived cardiomyocytes are induced PSC-derived cardiomyocytes.
15. The three-dimensional, synchronously contracting cardiac tissue of claim 13 , wherein the acrylate component comprises acrylated PEG.
16. The three-dimensional, synchronously contracting cardiac tissue of claim 13 , wherein the acrylate component comprises acrylated gelatin.
17. The three-dimensional, synchronously contracting cardiac tissue of claim 1 , wherein the hydrogel comprises a porogen.
18. The three-dimensional, synchronously contracting cardiac tissue of claim 1 , wherein the hydrogel comprises a matrix metalloproteinase degradable peptide.