CELL SUSPENSION AND USE THEREOF
The present invention provides for methods and devices suitable for producing a transplantable cellular suspension of living tissue suitable for promoting tissue regeneration in an epithelium-related procedure, as well as compositions produced therefrom. The cellular suspension can include viable and functioning cells at various stages of differentiation, including undifferentiated/progenitor cells and differentiated cells, as well as those in between. In certain embodiments, the cellular suspension can be subjected to a stress to induce a heat shock response therein, or be exposed to an exogenously supplied agent such as heat shock protein or a fragment thereof, hyaluronic acid, platelet-enriched plasma, and/or growth factors. The cellular suspension can be applied directly to a patient's recipient site for in vivo regeneration, or be cultured or seeded to a matrix for in vitro growth/regeneration.
1 . A method for preparing cells for use in an epithelium-related procedure, comprising:
preparing a suspension of cells from an epithelial tissue sample, wherein said cells are exposed to at least three stress conditions thereby inducing expression of a heat shock protein by the cells; and
applying the suspension of cells and the heat shock protein to a recipient site on a patient.
2 . The method of claim 1 , wherein the at least three stress conditions comprise mechanical disaggregation, enzymatic disaggregation, and a change in pH.
3 . The method of claim 2 , wherein the at least three stress conditions further comprise a change in osmotic pressure.
4 . The method of claim 2 , wherein the at least three stress conditions further comprise hypoxia.
5 . The method of claim 2 , wherein the at least three stress conditions further comprise calorie restriction.
6 . The method of claim 2 , wherein the at least three stress conditions further comprise a change in atmospheric pressure.
7 . The method of claim 2 , wherein the heat shock protein comprises at least one of Hsp90 and Hsp90a.
8 . The method of claim 7 , wherein the heat shock protein has a concentration between about 0.1 μg/μl to about 100 μg/μl.
9 . The method of claim 2 , wherein the suspension of cells comprises epidermal stem cells, melanocyte stem cells, follicular stem cells, and fibroblasts.
10 . The method of claim 2 , wherein the epithelial tissue sample is obtained from one or more of skin epithelium, respiratory epithelium, vascular epithelium, corneal epithelium, and glandular epithelium.
11 . The method of claim 2 , wherein applying the suspension of cells and the heat shock protein comprises applying a solution comprising the suspension of cells and the heat shock protein to the recipient site.
12 . The method of claim 11 , wherein applying the solution comprises at least one of spraying, dripping, spreading, pipetting, painting, and injecting the solution.
13 . The method of claim 12 , wherein applying the suspension of cells and the heat shock protein promotes at least one of epithelial treatment, healing, reconstructing, resurfacing, repigmentation, and regeneration in the patient.
14 . An apparatus for developing a tissue regeneration solution, comprising:
a first member comprising at least a heating element configured to heat an enzyme solution, a filter element configured to separate large cellular congregates, and a reservoir configured to hold a tissue sample immersed in a nutrient solution; and
a second member comprising at least one storage compartment, wherein the at least one storage compartment is configured to hold at least three tools and at least three vials,
wherein the apparatus is configured for use in preparing a suspension of cells from an epithelial tissue sample.
15 . The apparatus of claim 14 , wherein the at least three tools comprise at least three syringes.
16 . The apparatus of claim 14 , wherein the at least three vials comprise a vial that contains nutrient solution, a vial that contains enzyme solution, and a vial that contains sterile water.