Treatment of Diseases and Disorders Using Self-Renewing Colony Forming Cells Cultured and Expanded In Vitro

Abstract

The present invention relates to methods and uses of cells for the prevention and treatment of a wide variety of diseases and disorders and the repair and regeneration of tissues and organs using low passage and extensively passaged in vitro cultured, self-renewing, colony forming somatic cells (CF-SC). For example, adult bone marrow-derived somatic cells (ABM-SC), or compositions produced by such cells, are useful alone or in combination with other components for treating, for example, cardiovascular, neurological, integumentary, dermatological, periodontal, and immune mediated diseases, disorders, pathologies, and injuries.

1 . A method of administering a therapeutically useful amount of a biological composition or compositions to an organ, tissue, or subject, comprising administering to said organ, tissue, or subject an isolated population of bone marrow-derived self-renewing colony-forming somatic cells (CF-SC), wherein said CF-SC do not have multipotent differentiation capacity, wherein said CF-SC have a normal karyotype, wherein said CF-SC are non-immortalized, wherein said CF-SC express CD13, CD44, CD49c, CD90, HLA Class-1 and β (beta) 2-Microglobulin, and wherein said CF-SC do not express CD10, CD34, CD45, CD62L, or CD106. 
     
     
         2 . (canceled) 
     
     
         3 . A method of administering a therapeutically useful amount of a biological composition or compositions to an organ, tissue, or subject, comprising administering to said organ, tissue, or subject an isolated population of bone marrow-derived self-renewing colony-forming somatic cells (CF-SC), wherein said CF-SC do not have multipotent differentiation capacity, wherein said CF-SC have a normal karyotype, wherein said CF-SC are non-immortalized, and wherein said CF-SC are obtained from bone marrow by steps comprising: i) incubating bone marrow cells under a low oxygen condition such that said bone marrow cells when allowed to adhere to a tissue culture-treated surface produce adherent colony forming units; and, ii) passaging cells in said adherent colony forming units at low cell seeding densities. 
     
     
         4 . (canceled) 
     
     
         5 . The method of  claim 1 , wherein the therapeutically useful amount is an amount useful for, preventing tissue damage or for repairing, treating, or promoting regeneration of damaged tissue. 
     
     
         6 . (canceled) 
     
     
         7 . The method of  claim 3 , wherein the therapeutically useful amount is an amount useful for preventing tissue damage or for repairing, treating, or promoting regeneration of damaged tissue in an organ, tissue, or subject. 
     
     
         8 . (canceled) 
     
     
         9 . The method of  claim 1 , wherein the therapeutically useful amount is an amount useful for treating or reducing inflammation, immune, or autoimmune activity in an organ, tissue, or subject. 
     
     
         10 . (canceled) 
     
     
         11 . The method of  claim 3 , wherein the therapeutically useful amount is an amount useful for treating or reducing inflammation, immune, or autoimmune activity in an organ, tissue, or subject. 
     
     
         12 . (canceled) 
     
     
         13 . The method of  claim 1 , wherein said CF-SC further express one or more molecules selected from the group consisting of:
 a) CD29;   b) CD59;   c) CD147;   d) CD166; and,   e) telomerase   and, wherein said CF-SC further do not express one or more molecules selected from the group consisting of:   f) CD11c;   g) CD14;   h) CD33;   i) CD62P;   j) CD80;   k) STRO-1;   l) HLA-Class-II;   m) CD178.   n) p53; and,   o) p21.   
     
     
         15 . The method of  claim 3 , wherein said low oxygen condition is selected from the group consisting of
 a) less than about 20% oxygen;   b) less than about 15% oxygen;   c) less than about 10% oxygen;   d) less than about 5% oxygen;   e) between about 1 to 10% oxygen;   f) between about 2 to 7% oxygen;   g) between about 3 to 6% oxygen;   h) between about 4 to 6% oxygen; and,   i) between about 4 to 5% oxygen.   
     
     
         16 .- 19 . (canceled) 
     
     
         20 . The method of  claim 3 , wherein said low cell seeding density is selected from the group consisting of:
 a) less than about 2500 cells/cm 2 ;   b) less than about 1000 cells/cm 2 ; and,   c) less than about 500 cells/cm 2 .   
     
     
         21 .- 26 . (canceled) 
     
     
         27 . The method of  claim 1 , wherein said CF-SC have unipotent differentiation capacity. 
     
     
         28 . (canceled) 
     
     
         29 . The method of  claim 1 , wherein said CF-SC maintain an approximately constant population doubling rate through multiple in vitro cell doublings. 
     
     
         30 . (canceled) 
     
     
         31 . The method of  claim 1 , wherein said bone marrow is human bone marrow. 
     
     
         32 . The method of  claim 1 , wherein said cell population has undergone a number of population doublings selected from the group consisting of:
 a) at least about 15 population doublings;   b) at least about 20 population doublings;   c) at least about 25 population doublings;   d) at least about 30 population doublings;   e) at least about 35 population doublings;   f) at least about 40 population doublings;   g) at least about 45 population doublings; and,   h) at least about 50 population doublings.   
     
     
         33 .- 37 . (canceled) 
     
     
         38 . A composition comprising a pharmaceutically acceptable mixture of purified naturally occurring or isolated recombinant extracellular matrix or blood plasma proteins and bone marrow-derived self-renewing colony-forming somatic cells (CF-SC), wherein said CF-SC have a normal karyotype, wherein said CF-SC are non-immortalized, wherein said CF-SC express CD13, CD44, CD49c, CD90, HLA Class-1 and β (beta) 2-Microglobulin, and wherein said CF-SC do not express CD10, CD34, CD45, CD62L, or CD106. 
     
     
         39 . (canceled) 
     
     
         40 . A composition comprising a pharmaceutically acceptable mixture of purified naturally occurring or isolated recombinant extracellular matrix or blood plasma proteins and bone marrow-derived self-renewing colony-forming somatic cells (CF-SC), wherein said CF-SC have a normal karyotype, wherein said CF-SC are non-immortalized, and wherein said CF-SC are obtained from bone marrow by steps comprising: i) incubating bone marrow cells under a low oxygen condition such that said bone marrow cells when allowed to adhere to a tissue culture-treated surface produce adherent colony forming units; and, ii) passaging cells in said adherent colony forming units at low cell seeding densities. 
     
     
         41 .- 69 . (canceled) 
     
     
         70 . A method of inducing, enhancing, and/or maintaining the generation of new red blood cells in vitro or in vivo, wherein said induction, enhancement, or maintenance is achieved by co-cultivation of hematopoietic precursor cells with bone marrow-derived self-renewing colony-forming somatic cells (CF-SC), wherein said CF-SC have a normal karyotype, wherein said CF-SC are non-immortalized, wherein said CF-SC express CD13, CD44, CD49c, CD90, HLA Class-1 and β (beta) 2-Microglobulin, and wherein said CF-SC do not express CD10, CD34, CD45, CD62L, or CD106. 
     
     
         71 .- 72 . (canceled) 
     
     
         73 . A method of inducing, enhancing, and/or maintaining the generation of new red blood cells in vitro or in vivo, wherein said induction, enhancement, or maintenance is achieved by co-cultivation of hematopoietic precursor cells with bone marrow-derived self-renewing colony-forming somatic cells (CF-SC), wherein said CF-SC have a normal karyotype, wherein said CF-SC are non-immortalized, and wherein said CF-SC are obtained from bone marrow by steps comprising: i) incubating bone marrow cells under a low oxygen condition such that said bone marrow cells when allowed to adhere to a tissue culture-treated surface produce adherent colony forming units; and, ii) passaging cells in said adherent colony forming units at low cell seeding densities. 
     
     
         74 .- 94 . (canceled) 
     
     
         95 . The method of  claim 3 , wherein said CF-SC have unipotent differentiation capacity. 
     
     
         96 . The method of  claim 3 , wherein said CF-SC maintain an approximately constant population doubling rate through multiple in vitro cell doublings. 
     
     
         97 . The method of  claim 3 , wherein said bone marrow is human bone marrow. 
     
     
         98 . The method of  claim 3 , wherein said cell population has undergone a number of population doublings selected from the group consisting of:
 a) at least about 15 population doublings;   b) at least about 20 population doublings;   c) at least about 25 population doublings;   d) at least about 30 population doublings;   e) at least about 35 population doublings;   f) at least about 40 population doublings;   g) at least about 45 population doublings; and,   h) at least about 50 population doublings.