Composition and method for generating a desired cell type and/or tissue type from hair follicular stem cells

Abstract

The present invention is concerned with a composition and in vitro method for generating a desired cell type and/or tissue type from hair follicular stem cells. The composition and in vitro method are particularly suitable for generating an autologous desired cell type and/or tissue type. Furthermore, the composition and method are especially efficient and suitable for use in the context of cosmetic cell and/or tissue transplantation in recipient areas of a subject experiencing cell and/or tissue loss caused by, for example, a wound, scar, burn injury, tissue degeneration, and aging. The composition and in vitro method are also suitable to circumvent complications related to infections and/or immune rejection of a cosmetic cell and/or tissue implant or graft.

1 . A composition for generating a desired cell type and/or tissue type from at least one hair follicular stem cell, said composition comprising at least one anti-apoptotic compound, wherein said anti-apoptotic agent is a physiologically acceptable vanadium compound, at least one anti-oxidant compound, at least one stem cell enhancer compound, at least one extracellular matrix compound, and at least one differentiation inducing factor. 
     
     
         2 . The composition according to  claim 1 , wherein the physiologically acceptable vanadium compound is selected from the group consisting of bis (maltolato) oxovanadium, oxovanadium, and orthovanadium. 
     
     
         3 . The composition according to  claim 1 , wherein the physiologically acceptable vanadium compound is bis (maltolato) oxovanadium. 
     
     
         4 . The composition according to  claim 1 , further comprising one or more second or further anti-apoptotic compounds selected from the group of a triiodothyronine, estradiol, progesterone, tissue extract, insulin, transferrin, selenium, L-cysteine, adenosine triphosphate-magnesium chloride, and L-leucine. 
     
     
         5 . The composition according to  claim 4 , wherein the tissue extract is bovine pituitary extract. 
     
     
         6 . The composition according to  claim 1 , wherein the anti-oxidant compound is selected from the group of quercetin, monohydroxyethyl rutoside, vitamin C, lipoic acid, deferoxamine mesylate, and vitamin E. 
     
     
         7 . The composition according to  claim 6 , wherein the anti-oxidant compound is selected from quercetin and mono-hydroxyethyl rutoside. 
     
     
         8 . The composition according to  claim 1 , wherein the stem cell enhancer compound is selected from the group of: erythropoietin, CD34-positive cell, and retinoic acid. 
     
     
         9 . The composition according to  claim 8 , wherein the stem cell enhancer compound is selected from the group of erythropoietin and CD34-positive cell. 
     
     
         10 . The composition according to  claim 8 , wherein the stem cell enhancer compound is erythropoietin. 
     
     
         11 . The composition according to  claim 8 , wherein erythropoietin and/or the CD34-positive cell is derived from peripheral blood or bone marrow of a donor subject. 
     
     
         12 . The composition according to  claim 8 , wherein the CD34-positive cell is obtainable or obtained from a CD34-positive cell line. 
     
     
         13 . The composition according to  claim 8 , wherein the CD34-positive cell is obtainable or obtained from a human CD34-positive cell line. 
     
     
         14 . The composition according to  claim 1 , further comprising at least one degranulating agent. 
     
     
         15 . The composition according to  claim 14 , wherein the degranulating agent is Compound 48/80. 
     
     
         16 . The composition according to  claim 14 , further comprising an inorganic salt. 
     
     
         17 . The composition according to  claim 16 , wherein the inorganic salt is CaCl 2 . 
     
     
         18 . The composition according to  claim 1 , wherein the extracellular matrix compound is selected from the group of: platelet rich plasma, laminin, collagen IV, heparin sulfate, entactin, and chondroitin sulfate. 
     
     
         19 . The composition according to  claim 18 , wherein the extracellular matrix compound is platelet rich plasma. 
     
     
         20 . The composition according to  claim 19 , wherein the platelet rich plasma is derived from peripheral blood of a donor subject. 
     
     
         21 . The composition according to  claim 1 , further comprising one or more ingredients selected from the group of albumin, essential and non-essential amino acids, vitamins, trace elements, organic constituents, growth supplement, and antibiotics. 
     
     
         22 . The composition according to  claim 1 , wherein the desired cell type is selected from the group of nerve cell, photoreceptor cell, cardiomyocyte, odontoblast, epithelial cell, keratinocyte cell, fibroblast cell, fat cell, blood cell, immune cell, muscle cell, skin cell, hair follicular cell, osteoblast, osteocyte, osteoclast, and chondrocyte. 
     
     
         23 . The composition according to  claim 1 , wherein the desired tissue type is selected from the group of neural tissue, eye tissue, heart tissue, tooth tissue, epithelial tissue, keratinocyte tissue, fibroblast tissue, connective tissue, fat tissue, muscle tissue, skin tissue, hair tissue, bone tissue, and cartilage tissue. 
     
     
         24 . The composition according to  claim 1 , wherein the differentiation inducing factor is selected from the group consisting of basic fibroblast growth factor, insulin-growth factor, epidermal growth factor, transforming growth factor, nerve growth factor, fibroblast growth factor, epithelial growth factor, taurin, activin A, and 5-azacytidine. 
     
     
         25 . The composition according to  claim 1 , wherein the differentiation inducing factor is selected from basic fibroblast growth factor, insulin-growth factor, and nerve growth factor, and said desired cell type and/or tissue type is neural cell and/or neural tissue. 
     
     
         26 . The composition according to  claim 25 , wherein said insulin-like growth factor is selected from the group consisting of insulin-like growth factor type 1 and insulin-like growth factor type 2. 
     
     
         27 . The composition according to  claim 1 , wherein the differentiation inducing factor is epidermal growth factor, and said desired cell type and/or tissue type is epidermal cell and/or tissue type, skin cell and/or tissue type, blood vessel cell and/or tissue type, or bone cell and/or tissue type. 
     
     
         28 . The composition according to  claim 27 , wherein the epidermal growth factor is selected from the group consisting of: heparin-binding epidermal growth-like growth factor, epiregulin, epigen, betacellulin, neuregulin-1, neuregulin-2, neuregulin-3, and neuregulin-4. 
     
     
         29 . The composition according to  claim 1 , wherein the differentiation inducing factor is transforming growth factor, and said desired cell type and/or tissue type is cartilage cell and/or tissue type, heart valve cell and/or tissue type, and skeletal muscle cell and/or tissue type. 
     
     
         30 . The composition according to  claim 29 , wherein the transforming growth factor is selected from the group consisting of: transforming growth factor alpha and transforming growth factor beta. 
     
     
         31 . The composition according to  claim 1 , wherein the differentiation inducing factor is selected from activin A, taurin, and epidermal growth factor, and said desired cell type and/or tissue type is photoreceptor cell and/or retinal tissue. 
     
     
         32 . The composition according to  claim 1 , wherein the differentiation inducing factor is fibroblast growth factor, and said desired cell type and/or tissue type is fibroblast cell, skin cell and/or skin tissue, or blood vessel cell and/or blood vessel tissue. 
     
     
         33 . The composition according to  claim 1 , wherein the differentiation inducing factor is epithelial growth factor, and said desired cell type and/or tissue type is keratinocyte cell and/or keratinocyte tissue. 
     
     
         34 . The composition according to  claim 1 , which further comprises a serum free growth medium. 
     
     
         35 . An in vitro method for generating a desired cell type and/or tissue type from at least one hair follicular stem cell, said method comprising the steps of:
 (a) providing at least one hair follicular stem cell;   (b) culturing the at least one hair follicular stem cell in a medium so as to obtain a population of hair follicular stem cells;   (c) contacting the population of hair follicular stem cells of step (b) with a composition according to  claim 1  and allowing said population of hair follicular stem cells to differentiate into the desired cell type and/or tissue type under conditions favorable for differentiation; and   (d) harvesting the desired cell type and/or tissue type.   
     
     
         36 . The in vitro method according to  claim 35 , wherein the at least one hair follicular stem cell of step (a) is obtained from at least a part of a hair follicle in the anagen phase. 
     
     
         37 . The in vitro method according to  claim 36 , wherein the at least a part of a hair follicle in the anagen phase has been obtained by plucking a hair from a donor area of a subject. 
     
     
         38 . The in vitro method according to  claim 35 , wherein the at least a part of a hair follicle is contacted with a medium comprising collagenase IV. 
     
     
         39 . The in vitro method according to  claim 35 , wherein the hair follicular stem cell is a human hair follicular stem cell. 
     
     
         40 . The in vitro method according to  claim 35 , wherein the medium is a serum-free growth medium, optionally further supplemented with at least one tissue extract. 
     
     
         41 . The in vitro method according to  claim 40 , wherein the tissue extract is preferably a bovine pituitary extract compound. 
     
     
         42 . The in vitro method according to  claim 35 , which desired cell type and/or tissue type is intended for introduction in a recipient region in a subject, and wherein the at least one hair follicular stem cell of step (a) is derived from said subject, and wherein the composition of step (c) comprises at least one CD34-positive cell derived from said subject and/or platelet rich plasma derived from said subject. 
     
     
         43 . Use of a desired cell type and/or tissue type obtainable or obtained by the in vitro method according to  claim 35 , for a cosmetic purpose. 
     
     
         44 . Use of a desired cell type and/or tissue type obtainable by the in vitro method according to  claim 35 , for cosmetic tissue repair and/or wound healing. 
     
     
         45 . Use of a desired cell type and/or tissue type obtainable by the in vitro method according to  claim 35 , for cosmetic facial and body rejuvenation. 
     
     
         46 . Use of a desired cell type and/or tissue type obtainable by the in vitro method according to  claim 35  for regenerative medicine. 
     
     
         47 . A kit comprising the composition according to  claim 1 . 
     
     
         48 . The kit according to  claim 46  further comprising information on how to use the composition to generate a desired cell and/or tissue type.