Dental pulp marrow similar cells (dpmsc) and methods of isolating and using

Abstract

The invention provides for isolated population of pulp marrow similar cells (DPMSCs) and methods for isolating and using these cells. The population of DPMSCs are highly homogenous for CD1O, CD29, CD13, CD44, CD49a, CD49d, CD59, CD73, CDw90, CD105, Oct-4 Isoform A and B, Nanog, Sox-2, and SSEA-4.

1 . An isolated population of dental pulp marrow similar cells (DPMSC), wherein at least 90% of the cells in the population co-express of each of the following markers: CD10, CD13, CD29, CD44, CD49a, CD49d, CD59, CD73, CD90, CD105, Oct-4 Isoform A and B, Nanog, Sox-2, and SSEA-4. 
     
     
         2 . The population of  claim 1  wherein at least 95% of the cells in the population co-express each of the following markers: CD10, CD13, CD29, CD44, CD49a, CD49d, CD59, CD73, CD90, CD105, Oct-4 Isoform A and B, Nanog, Sox-2, and SSEA-4. 
     
     
         3 . The cell population of  claim 1  wherein about 90-99% of the cell population expresses each of the following markers: CD10, CD13, CD29, CD44, CD49a, CD49d, CD59, CD73, CD90, CD105, Oct-4 Isoform A and B, Nanog, Sox-2, and SSEA-4. 
     
     
         4 . The population of any one of  claims 1 - 3  wherein 0.25-1% of the cells in the population co-express CD34 and CD45. 
     
     
         5 . The population of any one of  claims 1 - 3  wherein the cells of the population co-express mRNA of each of the following: Oct-4 Isoforms A and B, Nanog, Sox-2, SSEA-4, c-Myc, Klf-4, and Rex-1. 
     
     
         6 . The population of any one of  claims 1 - 3  wherein the population has an average doubling rate of about 28-30 hours. 
     
     
         7 . The population of any one of  claims 1 - 3  wherein the DPMSCs have the capacity to differentiate into one or more of the cell types of ectodermal, endodermal, or mesodermal lineages. 
     
     
         8 . The population of any one of  claims 1 - 3  wherein the DPMSCs have the capacity to differentiate into at least two of the cell types of ectodermal, endodermal, or mesodermal lineages. 
     
     
         9 . The population of any one of  claims 1 - 3  wherein the DPMSCs have the capacity to differentiate into all three of the cell types of ectodermal, endodermal, or mesodermal lineages. 
     
     
         10 . The population of any one of  claims 1 - 3  wherein the DPMSCs are human DPMSCs. 
     
     
         11 . An isolated population of cells derived from human dental pulp, wherein at least 90% of the cells in the population co-express each of the following markers: CD10, CD29, CD13, CD44, CD49a, CD49d, CD59, CD73, CDw90, CD105, Oct-4, Nanog, Sox-2, and SSEA-4; wherein 0.25-1% of the cells in the population co-express CD34 and CD45; wherein the cells have normal karyotype; and wherein the cells of the population have the capacity to differentiate into cell types of at least two of ectodermal, endodermal, or mesodermal lineages. 
     
     
         12 . The population of any one of  claim 1 - 3  or  11  wherein the cells have the capacity to differentiate into any one or more of the following: osteoblast, skeletal muscle cell, smooth muscle cell, cardiac muscle cell, glial cell, and neuronal cell. 
     
     
         13 . The population of any one of  claim 1 - 3  or  11  wherein the DPMSCs are derived from a tooth organ. 
     
     
         14 . The cell population of  claim 12  wherein the tooth organ is from a child. 
     
     
         15 . The cell population of  claim 12  wherein the tooth organ is from an adult. 
     
     
         16 . An isolated population of cells which has been cultured to induce differentiation wherein the starting population is DPMSCs of  claim 1 , wherein the differentiation results in the DPMSC becoming a differentiated cell selected from a group consisting of: a bone cell, skeletal muscle cell, smooth muscle cell, cardiac muscle cell, glial cell, neuronal cell, skin epithelial cell, liver epithelial cell, pancreas epithelial cell, pancreas endocrine cell, pancreatic islet cell, pancreas exocrine cell, gut epithelium cell, kidney epithelium cell, epidermal associated structure, hair follicles, soft tissues surrounding teeth, dentin (teeth), enamel (teeth), and cement (teeth). 
     
     
         17 . The population of any one of  claim 1 - 3  or  11  wherein the genome of the cells in the population has not been altered by insertion of pre-selected isolated DNA, by substitution of a segment of the cellular genome with pre-selected isolated DNA, or by deletion of or inactivation of at least a portion of the cellular genome. 
     
     
         18 . A method of obtaining a population of DPMSCs of  claim 1  comprising culturing a dental pulp source in media supplemented with growth factors selected from the group consisting of: platelet-derived growth factor, insulin, selenium, epidermial growth factor (EGF), insulin-like growth factor (IGF), dexamethasone, linoleic acid, and ascorbic acid to obtain population of DPMSCs of  claim 1 . 
     
     
         19 . The method of  claim 18  wherein the DPMSCs are human DPMSCs and the dental pulp source is from a human. 
     
     
         20 . The method of  claim 18  wherein adherent cells and non-adherent cells are co-cultured without selection by immunodepletion, physical depletion, or chemical depletion. 
     
     
         21 . The method of  claim 20  wherein the method does not deplete the starting source of cells or the cell culture of mononuclear cells co-expressing CD3, CD14, CD19, CD38, CD66b and CD45 +  glycophorin A. 
     
     
         22 . The method  claim 20 , further comprising placing the cells in a cell culture container, wherein the cell culture container does not comprise an extracellular matrix (ECM) substrate. 
     
     
         23 . The method of  claim 20 , wherein serum percentage comprises about 0.5-2.25%. 
     
     
         24 . The method of  claim 18 , wherein the insulin is present at a concentration of about 10 to about 50 μg/ml, transferrin at a concentration of greater than 0 but less than about 10 μg/ml, selenium at a concentration of about 0.1 to about 5 μg/ml, linoleic acid at a concentration of about 0 to about 1 μg/m, dexamethasone at a concentration of about 0.005 to 0.15 μM, L-ascorbic acid at a concentration of about 10-50 mg F/L, platelet-derived growth factor at a concentration of about 5 to about 15 ng/ml, epidermal growth factor 1 to at a concentration of about 15 ng/ml, insulin-like growth factor at a concentration of 1 to about 15 ng/ml, and ml and fibroblast growth factor-b1 to at a concentration of about 15 ng/ml. 
     
     
         25 . A method of providing therapeutic assistance to an individual in need thereof comprising administering an amount of DPMSCs effective to aid therapy to the individual. 
     
     
         26 . A population of DPMSCs produced by the culturing method of any one of  claims 18 - 24 .