Human uterine cervical stem cell population and uses thereof

Abstract

The present invention relates to a method for isolating stem cells comprising preparing a cell suspension from uterine cervix tissue, to the stem cells isolated by said method, and to the conditioned medium obtained from the culture of said stem cells. The invention also encompasses the use of said stem cells or conditioned medium for treating or preventing cancer, precancerous lesions, inflammatory diseases, autoimmune diseases, chronic pathologies or infectious diseases, diseases associated to tissue loss, or for use in diagnostic, prognostic or treatment of fertility disorders, as well as for cosmetic treatment.

1 . A method for isolating stem cells comprising:
 (a) Preparing a cell suspension from uterine cervix tissue,   (b) Recovering the cells from said cell suspension,   (c) Incubating said cells in a suitable cell culture medium and under conditions which allow cells to proliferate, and   (d) Selecting the stem cells.   
     
     
         2 . Method according to  claim 1 , wherein the step (a) comprises enzymatically disaggregating the cervical mucus. 
     
     
         3 . Method according to  claim 1  or  2 , wherein the isolated stem cells:
 (a) express the cell markers CD29, CD44, CD73, CD90, CD105, vimentin, cytokeratin (CKAE1AE3), Klf4, Oct4 and Sox-2, and 
 (b) do not express at least one cell marker selected from the group consisting of desmin, actin HHF35, β-catenin, p63, E-cadherin, CD117, CD133, HLA-DR, TRA1-81, CD45, CD34 and CD31. 
 
     
     
         4 . Method according to  claim 3 , wherein the stem cells further show
 (a) a proliferating rate from 0.4 to 2.1 doublings per 24 hours in growth medium,   (b) a fibroblast-like morphology,   (c) a stable karyotype for at least 10 cell passages,   (d) capacity to grow in monolayer and to adhere to a substrate,   (e) capacity to be differentiated into endodermal, ectodermal or mesodermal cell lineage,   (f) a non tumorigenic capacity and/or   (g) capacity to form spheres.   
     
     
         5 . Method according to any one of  claims 1  to  4 , wherein the uterine cervix tissue is a mammalian uterine cervix tissue, preferably, a human uterine cervix tissue. 
     
     
         6 . Method according to any one of  claims 1  to  5 , wherein the uterine cervix tissue is a non-cancerous tissue. 
     
     
         7 . Use of isolated uterine cervix tissue according to any one of  claims 1  to  6 , for obtaining uterine cervix stem cells. 
     
     
         8 . Use according to  claim 1  wherein the uterine cervix tissue is non-cancerous. 
     
     
         9 . An isolated uterine cervix stem cell, wherein said cell:
 (a) expresses the cell markers CD29, CD44, CD73, CD90, CD105, vimentin, cytokeratin (CKAE1AE3), Klf4, Oct4 and Sox-2, and   (b) does not express at least one cell marker selected from the group consisting of desmin, actin HHF35, β-catenin, p63, E-cadherin, CD117, CD133, HLA-DR, TRA1-81, CD45, CD34 and CD31.   
     
     
         10 . Isolated stem cell according to  claim 9 , wherein the cell further shows
 (a) a proliferating rate from 0.4 to 2.1 doublings per 24 hours in growth medium,   (b) a fibroblast-like morphology,   (c) a stable karyotype for at least 10, preferably, 20 cell passages,   (d) capacity to grow in monolayer and to adhere to a substrate,   (e) capacity to be differentiated into an adipogenic, osteogenic, neural or myocytic cell linage,   (f) a non tumorigenic capacity and/or   (g) capacity to form spheres.   
     
     
         11 . Isolated stem cell according to  claim 9  or  10 , wherein the cell is from a mammal, preferably, from a human, more preferably, from a human in a non-menstrual phase. 
     
     
         12 . A cell population comprising an isolated stem cell according to any one of  claims 9  to  11 . 
     
     
         13 . A conditioned medium obtained by a method comprising:
 (a) Incubating an isolated stem cell according to any one of  claims 9  to  11  or a cell population according to  claim 12 , and   (b) Removing the cells from the culture medium.   
     
     
         14 . A pharmaceutical composition comprising an isolated stem cell according to any one of  claims 9  to  11 , a cell population according to  claim 12 , or a conditioned medium according to  claim 13 , and an acceptable pharmaceutically carrier and/or an adjuvant. 
     
     
         15 . An isolated stem cell according to any one of  claims 9  to  11 , a cell population according to  claim 12 , a conditioned medium according to  claim 13  or a pharmaceutical composition according to  claim 14  for use as a medicament. 
     
     
         16 . An isolated stem cell according to any one of  claims 9  to  11 , a cell population according to  claim 12 , a conditioned medium according to  claim 13  or a pharmaceutical composition according to  claim 14 , for use in the treatment or prevention of cancer, precancerous lesions, inflammatory diseases, autoimmune diseases, chronic pathologies or infectious diseases, diseases associated to tissue loss, or for use in diagnostic, prognostic or treatment of fertility disorders or for use in a cosmetic treatment. 
     
     
         17 . An isolated stem cell according to any one of  claims 9  to  11 , a cell population according to  claim 12 , a conditioned medium according to  claim 13  or a pharmaceutical composition according to  claim 14 , for inhibiting or decreasing the proliferation and/or metastasis of tumor cells, the monocytic differentiation, the peripheral blood mononuclear cells proliferation or the pathogenic microorganism growth and/or replication, or for enhancing or inducing the apoptosis of tumor cells, or for enhancing tissue regeneration and selection of germ cells. 
     
     
         18 . A kit comprising an isolated stem cell according to any one of  claims 9  to  11 , a cell population according to  claim 12 , or a conditioned medium according to  claim 13 , or the pharmaceutical composition according to  claim 14 . 
     
     
         19 . Use of the kit according to  claim 18  for the treatment or prevention of cancer, precancerous lesions, inflammatory diseases, autoimmune diseases, chronic pathologies or infectious diseases, diseases associated to tissue loss, or for use in diagnostic, prognostic or treatment of fertility disorders or for use in a cosmetic treatment. 
     
     
         20 . Use of the kit according to  claim 18  for inhibiting or decreasing the proliferation and/or metastasis of tumor cells, the monocytic differentiation or peripheral blood mononuclear cells proliferation, or for enhancing or inducing the apoptosis of tumor cells, or for enhancing tissue regeneration, or for the selection of germ cells.