BIOACTIVE RENAL CELLS
The present invention concerns bioactive renal cell populations, renal cell constructs, and methods of making and using the same.
1 - 34 . (canceled)
35 . A method of providing a regenerative effect to a native kidney comprising in vivo contacting the native kidney with a composition comprising isolated human secreted vesicles produced by a renal cell population enriched for bioactive kidney cells,
wherein the vesicles comprise exosomes or microvesicles comprising a paracrine factor that attenuates Nuclear Factor kappa B (NFκB) signaling,
wherein the vesicles have been isolated from the renal cell population, and
wherein the regenerative effect comprises a reduction in renal inflammation.
36 . The method of claim 35 , wherein the vesicles comprise microvesicles.
37 . The method of claim 35 , wherein the vesicles comprise exosomes.
38 . The method of claim 35 , wherein the paracrine factor comprises at least one miRNA that inhibits NFκB.
39 . The method of claim 38 , wherein the at least one miRNA comprises miR-146a, miR-124, or miR-151.
40 . The method of claim 39 , wherein the at least one miRNA further comprises miR-30b-5p, miR449a, miR130a, miR-23b, or miR-21.
41 . The method of claim 35 , wherein the bioactive kidney cells are tubular cells.
42 . The method of claim 41 , wherein the renal cell population further comprises epithelial cells of the collecting duct.
43 . The method of claim 35 , wherein the renal cell population is autologous to the native kidney.
44 . The method of claim 35 , wherein the renal cell population is non-autologous to the native kidney.
45 . An in vitro method of screening a test article comprising an enriched renal cell population for biotherapeutic efficacy, the method comprising:
culturing the test article;
collecting conditioned media from the cultured test article; and
determining whether the conditioned media attenuates nuclear factor kappa B (NFκB) or plasminogen activator inhibitor (PAI)-1 signaling,
wherein the enriched renal cell population is screened as having biotherapeutic efficacy if it attenuates NFκB or PAI-1 signaling, and
wherein the biotherapeutic efficacy comprises attenuation of an immune or fibrotic response.
46 . The in vitro method of claim 45 , wherein the determining determines whether the conditioned media attenuates PAI-1 signaling in cells.
47 . The in vitro method of claim 46 , wherein the cells are mesangial cells.
48 . The in vitro method of claim 45 , wherein the determining determines whether the conditioned media attenuates NFκB signaling.
49 . The in vitro method of claim 48 , wherein the determining determines whether the conditioned media attenuates NFκB signaling in cells.
50 . The in vitro method of claim 49 , wherein the cells are proximal tubule cells.
51 . The in vitro method of claim 50 , wherein the proximal tubule cells are HK-2 cells.
52 . The in vitro method of claim 49 , wherein the conditioned media is determined to attenuate NFκB signaling in the cells if it reduces tumor necrosis factor (TNF)-α induced nuclear localization of NFκB subunit p65.