INJECTABLE CELL AND SCAFFOLD COMPOSITIONS
Provided herein are, inter alia, therapeutic formulations containing active agents, such as bioactive cell populations, and methods of making and using the same.
1 . An injectable formulation comprising:
a) a temperature-sensitive cell-stabilizing biomaterial, and
b) a bioactive renal cell (BRC) population,
wherein the temperature-sensitive cell-stabilizing biomaterial is a hydrogel that
(i) maintains a substantially solid state at about 8° C. or below, wherein the substantially solid state is a gel state,
(ii) maintains a substantially liquid state at about ambient temperature or above, and
(iii) has a solid-to-liquid transitional state between about 8° C. and about ambient temperature or above,
wherein the hydrogel comprises an extracellular matrix protein of recombinant origin, is derived from extracellular matrix sourced from kidney or another tissue or organ, or comprises gelatin.
2 . The injectable formulation of claim 1 , wherein the gelatin is derived from Type I, alpha I collagen.
3 - 5 . (canceled)
6 . The injectable formulation of claim 1 , wherein the BRC is a selected renal cell (SRC) population.
7 . (canceled)
8 . The injectable formulation of claim 6 , wherein the BRC or SRC population is enriched for tubular renal cells.
9 - 10 . (canceled)
11 . The injectable formulation of claim 8 , wherein the SRC population is characterized by phenotypic expression of one or more tubular epithelial cell markers, wherein the one or more tubular epithelial cell markers comprise CK18 and/or GGT1.
12 - 13 . (canceled)
14 . The injectable formulation of claim 8 , wherein the SRC population is characterized by phenotypic expression of one or more viability and/or functionality markers, and wherein the one or more viability and/or functionality markers comprise VEGF and/or KIM-1.
15 . The injectable formulation of claim 8 , wherein the SRC population is characterized by LAP and/or GGT enzymatic activity.
16 - 20 . (canceled)
21 . An implantable formulation comprising:
a) a decellularized kidney of human or animal origin or a cell-stabilizing biomaterial that has been structurally engineered through three dimensional bioprinting, and
b) a bioactive renal cell (BRC) population.
22 . (canceled)
23 . The injectable formulation of claim 1 , wherein:
a) the biomaterial comprises gelatin at about 0.88% (w/v), and wherein the gelatin is derived from Type I, alpha I collagen, and
b) the BRC population comprises a selected renal cell (SRC) population, wherein the SRC population comprises an enriched population of tubular renal cells and has a density greater than about 1.04 g/mL.
24 . A method for preparing an injectable formulation comprising a temperature-sensitive cell-stabilizing biomaterial and an admixture of bioactive renal cells, the method comprising the steps of: i) obtaining renal cortical tissue from the donor/recipient; ii) isolating renal cells from the kidney tissue by enzymatic digestion and expanding the renal cells using standard cell culture techniques; iii) subjecting the harvested renal cells to separation across a density boundary or density interface or single step discontinuous gradient to obtain an SRC population; and iv) reconstituting the bioactive cells with a gelatin-based hydrogel biomaterial, wherein the gelatin is derived from Type I, alpha I collagen.
25 . The method of claim 24 , wherein the selected renal cells comprise an enriched population of tubular renal cells and having a density greater than about 1.04 g/mL.
26 . The method of claim 24 , wherein the harvested renal cells are exposed to hypoxic culture conditions prior to separation across a density boundary or density interface or continuous or discontinuous single step or multistep density gradient.
27 . The method of claim 24 , wherein the renal cells are enriched for tubular renal cells.
28 - 32 . (canceled)
33 . The method of claim 24 , further comprising monitoring the renal cells for phenotypic expression of one or more viability and/or functionality markers, wherein the one or more viability and/or functionality markers comprise VEGF and/or KIM-1.
34 . (canceled)
35 . The method of claim 24 , further comprising monitoring the renal cells for phenotypic expression of one or more tubular epithelial cell markers, wherein the one or more tubular epithelial cell markers comprise CK18 and/or GGT1.
36 . (canceled)
37 . The method of claim 24 , further comprising monitoring renal cell functionality by measuring enzymatic activity, wherein the measured enzymatic activity is for LAP and/or GGT.
38 - 40 . (canceled)
41 . The method of claim 24 , wherein the SRC are resuspended in a liquefied gelatin solution at 26-30° C.
42 . The method of claim 41 , wherein the SRC are resuspended in sufficient gelatin solution to achieve an SRC concentration of 100×10 6 cells/ml.
43 . The method of claim 24 , further comprising rapidly cooling the SRC/gelatin solution to stabilize the biomaterial such that the SRC will remain suspended in the gel on storage.
44 - 47 . (canceled)
48 . A method of treating kidney disease in a subject, the method comprising injecting the formulation of claim 1 into the subject, wherein the formulation is injected through a 18 to 30 gauge needle.
49 . (canceled)