Placenta-derived matrix and methods of preparing and use thereof
The invention relates to placenta-derived matrix, methods of preparing, and methods of use thereof. The invention also relates to methods of culturing cells, delivering cells, promoting differentiation of stem cells or tissue-specific progenitor cells, and repairing, replacing, regenerating, filling, reducing or inhibiting scarring of defects using the same. The invention further relates to methods of coating placenta-derived matrix on a surface or injecting the placenta-derived matrix into a site of interest.
1. A method of preparing a hydrogel obtained from a placental tissue, said method comprising
(a) devitalizing a placenta tissue to produce a devitalized placenta tissue,
(b) digesting the devitalized placenta tissue in a digestion solution having a pepsin and hydrochloric acid (HCl) to produce a placenta-derived matrix,
(c) adjusting pH of the placenta-derived matrix from step (b) to a basic pH of 8.0 to 11.0 to irreversibly inactivate the pepsin,
(d) adjusting the pH of the placenta-derived matrix from step (c) to a neutral pH of 7.0 to 7.2 to prepare a first neutralized placenta-derived matrix,
(e) freeze-drying the first neutralized placenta-derived matrix to produce a sponge structure,
(f) dissolving the sponge structure in an acetic acid solvent to form a second placenta-derived matrix,
(g) adjusting the pH of the second placenta-derived matrix from step (f) to a neutral pH of 7.0 to 7.2 to prepare a second neutralized placenta-derived matrix, and
(h) adjusting the temperature of the second neutralized placenta-derived matrix to 36 to 38° C. to produce the hydrogel.
2. The method of claim 1 , wherein step (a) comprises treating the placenta tissue with a non-denaturing detergent.
3. The method of claim 1 , further comprising homogenizing the devitalized placenta tissue concurrently with or before digesting the devitalized placenta tissue of step (b).
4. The method of claim 3 , wherein the devitalized placenta tissue is homogenized at 4° C.
5. The method of claim 1 , wherein the concentration of pepsin in the digestive solution is from 400 to 700 units/ml, and the concentration of HCl in the digestive solution is 0.01M-1.0M.
6. The method of claim 1 , further comprising storing cells on or in the hydrogel.
7. The method of claim 6 , wherein the cells on or in the hydrogel is stored by cryopreservation.
8. The method of claim 1 , further comprising exposing a protein to the hydrogel.
9. The method of claim 1 , further comprising
(a) mixing cells in the hydrogel to produce a mixture, and
(b) injecting the mixture into a site of interest.
10. The method of claim 1 , further comprising culturing pluripotent stem cells or tissue-specific progenitor cells on or within the hydrogel.
11. The method of claim 1 , further comprising coating at least a part of a surface of a substrate with the hydrogel.