Methods of enhancing fibroblast therapeutic activity
Disclosed are compositions of matter, cells, protocols and procedures useful for augmentation of one or more therapeutic activities of fibroblast cellular populations. In one embodiment fibroblasts are pretreated with growth factor-comprising composition(s), wherein the growth factor(s) may be cytokines, peptides, and/or proteins. In another embodiment fibroblasts are cultured with platelet rich plasma and/or derivatives from platelet rich plasma. In another embodiment, fibroblasts are cultured under hypoxic conditions prior to administration to an individual. The disclosure further provides means of assessment of fibroblast activity in vitro, including wound repair assay and cytokine production, for example.
1. A method of generating regenerative fibroblasts comprising the step of contacting fibroblasts with platelet-rich plasma and one or more biologically active substances, thereby generating regenerative fibroblasts having improved anti-inflammatory activity compared to fibroblasts that lack said contact, wherein the one or more biologically active substances comprise hyaluronic acid and batroxobin.
2. The method of claim 1 , wherein said biologically active substance comprises one or more cytokines.
3. The method of claim 2 , wherein said cytokine comprises one or more growth factors.
4. The method of claim 3 , wherein said growth factor is FGF-alpha.
5. The method of claim 3 , wherein said growth factor is FGF-beta.
6. The method of claim 3 , wherein said growth factor is a member of the TGF-beta family.
7. The method of claim 1 , wherein the fibroblast cells are selected from the group consisting of (a) fibroblasts obtained by biopsy, cultured and proliferated; (b) subsets thereof having greater ability to differentiate; and (c) a combination thereof.
8. The method of claim 1 , wherein the fibroblasts express stage specific embryonic antigen 3 (SSEA3).
9. The method of claim 1 , wherein said fibroblasts are comprised in a pharmaceutically acceptable carrier selected from the group consisting of sterile solutions, hydrogels, implantable cell matrices, devices and a combination thereof.
10. The method of claim 1 , wherein the fibroblasts are derived from tissues comprising skin, heart, blood vessels, bone marrow, skeletal muscle, liver, pancreas, brain, adipose tissue, foreskin, placental, and/or umbilical cord.
11. The method of claim 1 , wherein the fibroblasts are placental, fetal, neonatal, adult or a combination thereof.
12. The method of claim 1 , wherein a therapeutically effective amount of the fibroblasts are provided to the disc of an individual in need of disc repair.
13. The method of claim 1 , wherein the fibroblasts are recombinantly manipulated to encode SSEA3, VEGF, FGF-1, FGF-2, FGF-4, EGF, HGF, HIF-1alpha, HIF-2, NET, NF-kB, TGF-a, TGF-b, IL-4, IL-10, IL-13, IL-20 thrombospondin, canstatin, IP-10, kringle 1-5, collagen XVIII/endostatin, IL-1, TNF-alpha, IL-2, IL-6, IL-8, IL-9, IL-11, IL-12, IL-15, IL-17, IL-18, IL-21, IL-23, IL-27, IFN-alpha, IFN-beta, IFN-gamma, p55, p60, STAT1, STATS, STAT4, IRF-1, IFR-3, IFR-8, or a combination thereof.