Amnion-derived cell compositions, methods of making and uses thereof

Abstract

The invention is directed to substantially purified amnion-derived cell populations, compositions comprising the substantially purified amnion-derived cell populations, and to methods of creating such substantially purified amnion-derived cell populations, as well as methods of use. The invention is further directed to antibodies, in particular, monoclonal antibodies, that bind to amnion-derived cells or, alternatively, to one or more amnion-derived cell surface protein markers. The invention is further directed to methods for producing the antibodies, methods for using the antibodies, and kits comprising the antibodies.

1 . A substantially purified population of amnion-derived cells that is negative for expression of the protein markers CD90 and CD117.  
   
   
       2 . The substantially purified population of  claim 1  that is further negative for expression of the protein marker CD105.  
   
   
       3 . The substantially purified population of  claim 1  that is positive for expression of the protein marker CD29.  
   
   
       4 . The substantially purified population of  claim 3  that is negative for expression of the protein marker CD105.  
   
   
       5 . The substantially purified population of  claim 3  that is further positive for expression of at least one of the protein markers selected from the group consisting of CD9, CD10, CD26, CD71, CD166, CD227, EGF-R, SSEA-4, and HLA-G.  
   
   
       6 . The substantially purified population of  claim 4  that is further positive for expression of at least one of the protein markers selected from the group consisting of CD9, CD10, CD26, CD71, CD166, CD227, EGF-R, SSEA-4, and HLA-G.  
   
   
       7 . The substantially purified population of  claim 2  that is further negative for the expression of at least one of the protein markers selected from the group consisting of CD140b, telomerase, CD34, CD44, and CD45.  
   
   
       8 . The substantially purified population of  claim 4  that is further negative for the expression of at least one of the protein markers selected from the group consisting of CD140b, telomerase, CD34, CD44, and CD45.  
   
   
       9 . The substantially purified population of  claim 6  that is further negative for the expression of at least one of the protein markers selected from the group consisting of CD140b, telomerase, CD34, CD44, and CD45.  
   
   
       10 . The composition of  claim 1 , which is a pharmaceutical composition.  
   
   
       11 . A method of obtaining the substantially purified population of amnion-derived cells of  claim 1 , comprising: 
 a) providing a population of amnion-derived cells;    b) contacting the cells with 
 (i) one or more antibodies selected from the group consisting of anti-CD105, anti-CD90, anti-CD117, anti-CD140b, anti-CD34, anti-CD44, and anti-CD45, antibodies; and  
 (ii) one or more antibodies selected from the group consisting of anti-CD29, anti-CD9, anti-CD10, anti-CD26, anti-CD71, anti-CD166, anti-CD227, anti-EGF-R, anti-SSEA-4, and anti-HLA-G antibodies; and  
   c) separating the cells that do not bind to the antibodies of (i) from the cells that do bind to the antibody of (i) and separating the cells that do not bind to the antibodies of (ii) from the cells that do bind to the antibody of (ii) such that a substantially purified population of amnion-derived cells that do not bind to the antibodies of (i) and do bind to the antibody of (ii) is obtained.    
   
   
       12 . A method of obtaining a substantially purified population of amnion-derived cells, comprising: 
 a) providing a population of amnion-derived cells;    b) contacting the cells with 
 (i) one or more antibodies selected from the group consisting of anti-CD105, anti-CD90, anti-CD117, anti-CD140b, anti-CD34, anti-CD44, and anti-CD45, antibodies; and  
 (ii) one or more antibodies selected from the group consisting of anti-CD29, anti-CD9, anti-CD10, anti-CD26, anti-CD71, anti-CD166, anti-CD227, anti-EGF-R, anti-SSEA-4, and anti-HLA-G antibodies; and  
   c) separating the cells that do not bind to the antibodies of (i) from the cells that do bind to the antibody of (i) and separating the cells that do not bind to the antibodies of (ii) from the cells that do bind to the antibody of (ii) such that a substantially purified population of amnion-derived cells that do not bind to the antibodies of (i) and do bind to the antibody of (ii) is obtained.    
   
   
       13 . The amnion-derived cells of  claim 1 , which are an expanded amnion-derived cell composition.  
   
   
       14 . The composition of  claim 13  which is animal-free.  
   
   
       15 . The composition of  claim 13  wherein the amnion-derived cells form spheroids.  
   
   
       16 . A composition comprising conditioned medium obtained from the expanded amnion-derived cell composition of  claim 13 .  
   
   
       17 . A composition comprising cell lysate obtained from the amnion-derived cell composition of  claim 13 .  
   
   
       18 . The expanded amnion-derived cell composition of  claim 13  having a concentration of at least 500×10 6  amnion-derived cells/g of starting amnion.  
   
   
       19 . A method of creating a hepatocyte comprising differentiating, in vitro or in vivo, an amnion-derived cell population of  claim 1 .  
   
   
       20 . A hepatocyte created by the method of  claim 19 .  
   
   
       21 . A method of creating a cardiomyocyte comprising differentiating, in vitro or in vivo, an amnion-derived cell population of  claim 1 .  
   
   
       22 . A cardiomyocyte created by the method of  claim 21 .  
   
   
       23 . A method for promoting accelerated wound healing in an injured patient in need thereof comprising administering to the patient one or more compositions selected from the group consisting of placental-derived cells, conditioned media derived from placental-derived cells, placental-derived cell lysates, and placental-derived cell products.  
   
   
       24 . The method according to  claim 23  wherein the wound is selected from the group consisting of mechanical, thermal, acute, chronic, infected, and sterile wounds.  
   
   
       25 . The method according to  claim 23  wherein the injured patient is a human.  
   
   
       26 . A cosmetic preparation comprising one or more compositions selected from the group consisting of placental-derived cells, conditioned media derived from placental-derived cells, placental-derived cell lysates, and placental-derived cell products.  
   
   
       27 . A method for treating hearing loss in a patient in need thereof comprising administering to the patient one or more compositions selected from the group consisting of placental-derived cells, conditioned media derived from placental-derived cells, placental-derived cell lysates, and placental-derived cell products.  
   
   
       28 . The population of  claim 1 , wherein the cells express a pancreatic progenitor cell marker protein.  
   
   
       29 . The population of  claim 28 , wherein the progenitor cell marker is PDX1 protein and wherein the cells further optionally express any one or more of the protein markers selected from the group consisting of Foxa2, p48, Hblx9, Neurogenin 3 (Hgn3), NKx2.2, Nkx6.1, insulin and islet-1.  
   
   
       30 . The population of  claim 29 , wherein the PDX1 protein is expressed in the nucleus.  
   
   
       31 . The population of  claim 28 , wherein the cells are differentiated pancreatic progenitor cells.  
   
   
       32 . The population of  claim 31 , wherein the differentiated progenitor cells express any one or more of the protein markers selected from the group consisting of PDX1, insulin, C-peptide, somatostatin, pancreatic polypeptide, and glucagon.  
   
   
       33 . The population of  claim 31 , wherein the differentiated pancreatic progenitor cells are islet-like cells.  
   
   
       34 . The population of  claim 33 , wherein the islet-like cells are functional alpha, beta, delta or phi cells.  
   
   
       35 . The population of  claim 34 , wherein functionality of the islet-like cells is incremental glucose-dependent insulin secretion.  
   
   
       36 . An islet comprising the population of  claim 28 .  
   
   
       37 . A tissue comprising the population of  claim 28 .  
   
   
       38 . The population of  claim 28 , wherein the cells form spheroids.  
   
   
       39 . The population of  claim 38 , wherein the spheroids form buds.  
   
   
       40 . The population of  claim 39 , wherein the buds express PDX1 protein.  
   
   
       41 . The population of  claim 40 , wherein the PDX1 protein is expressed in the nucleus.  
   
   
       42 . The population of  claim 28 , which comprises one or more mammalian embryonic islet progenitor cells.  
   
   
       43 . The population of  claim 42 , wherein the mammalian embryonic islet progenitor cells are human cells.  
   
   
       44 . The population of  claim 28 , wherein the cells express a heterologous protein.  
   
   
       45 . The population of  claim 28 , wherein the cells have the identifying characteristics of endoderm, wherein the identifying characteristics of endoderm are expression of HNF1α, HNF1β, HNF4α, HNF6, Foxa2 and PDX1 proteins and wherein the cells further optionally expressing any one or more of the protein markers Sox17, Cerberus, Hesx1, LeftyA, Otx1 or Otx2.  
   
   
       46 . A composition comprising one or more nuclei isolated from pancreatic progenitor cells of  claim 28 , wherein the cells express PDX1 protein in the nucleus and/or express Nkx2.2, Nkx6.1, insulin and islet-1 protein and/or have the identifying characteristics of endoderm.  
   
   
       47 . The composition of claim  4469 , wherein the identifying characteristics of endoderm are protein expression of HNF1α, HNF1β, HNF4α, HNF-6, Foxa2 and PDX1 and wherein the cells, further optionally express any one or more of the protein markers Sox17, Cerberus, Hesx1, LeftyA, Otx1 or Otx2.  
   
   
       48 . A pharmaceutical composition comprising an effective amount of the population of  claim 28  and a carrier.  
   
   
       49 . An in vivo method for inducing differentiation of resident pancreatic cells into islet cells comprising: 
 a) introducing factors into the pancreas of a subject; and    b) allowing the introduced factors to prime the resident pancreatic cells such that the cells are induced to differentiate into islet progenitor cells and/or islet cells.    
   
   
       50 . An in vivo method for promoting the generation of islet cells in a subject comprising: 
 a) transplanting the population of amnion-derived cells of  claim 1  into the pancreas of the subject:    b) introducing factors into the pancreas of the subject; and    c) allowing the introduced factors to promote generation of islet progenitor cells and or islet cells from the transplanted amnion-derived cells.    
   
   
       51 . An in vivo method for promoting the differentiation of amnion-derived cells into pancreatic cells comprising 
 (a) co-culturing the population of amnion-derived cells of  claim 1  with differentiating embryonic pancreatic or non-pancreatic tissue; and    (b) transplanting the co-cultures into the pancreas of a subject.    
   
   
       52 . An in vivo method for promoting the differentiation of amnion-derived cells into pancreatic cells comprising 
 (a) co-culturing the population of amnion-derived cells of  claim 1  with differentiating or pre-differentiating non-embryonic heterologous tissue or autologous tissue; and    (b) transplanting the co-cultures into the pancreas of a subject.    
   
   
       53 . An in vivo method for promoting the differentiation of amnion-derived cells into pancreatic cells comprising 
 (a) introducing factors to the population of amnion-derived cells of  claim 1  in vitro; and    (b) subsequently transplanting the amnion-derived cells into the pancreas of a subject.    
   
   
       54 . A cell culture system comprising a cell culture medium comprising a SHh antagonist and the population of  claim 1  or  claim 13 .  
   
   
       55 . A cell comprising a nucleus isolated from the amnion-derived cell of  claim 1.