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

a technology of dental pulp and similar cells, which is applied in the field of compositions and methods for isolating and using dental pulp marrow similar cells (dpmscs), can solve the problems of significant cost, both financially and physically, for the patient to receive anti-rejection medications at a significant cost, and achieve the effect of easing cardiac failur

Inactive Publication Date: 2011-06-30
FOUND OF TRANSLATIONAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0046]In yet another aspect, the invention provides methods for providing therapeutic assistance to treat cardiac diseases in an individual in need thereof comprising administering to the individual an amount of cardiomyocytes derived from DPMSCs effect

Problems solved by technology

However, there are at least two major problems associated with organ and tissue transplantation.
First, there is a shortage of donor organs and tissues.
The second major problem is the potential incompatibility of the transplanted tissue with the immune system of the recipient.
Because the donated organ or tissue is recognized by the host immune system as foreign, anti-rejection medications must be provided to the patient at a significant cost, both financially and physically.
Unfortunately, xenotransplantation does not overcome the problem of tissue incompatibility, but even exacerbates it.
Furthermore, according to the Centers for Disease Control, there is evidence that damaging viruses cross species barriers.
Stem cells which differentiate only to form cells of hematopoietic lineage, however, are unable to provide a source of cells for repair of other damaged tissues, for example, heart or lung tissue damaged by high-dose chemotherapeutic agents.
This cell is capable of differentiating to form a number of cell types of mesenchymal origin, but has been reported by the team who isolated it to be apparently limited in differentiation potential to cells of the mesenchymal lineage since hematopoietic cells were never identified in the expanded cultures.
In addition, the degree of telomerase activity in tissue specific

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1a

Isolation of DPMSC

[0223]Cell Culture

[0224]Dental pulp was extracted from normal exfoliated human deciduous teeth of 5- to 7-year old children (10 patients) with informed consent of the donors. Dental pulp was extracted using needle and was transferred into 35-mm Petri dishes F12 / Medium 199 / CMRL 1066 supplemented with 1.25% Human serum and supplemented with 1-50 ng / ml (preferably about 5-15 ng / ml) platelet derived growth factor-BB (PDGF-BB), 1-50 ng / ml (preferably about 1-15 ng / ml) epidermal growth factor (EGF), 1-50 ng / ml (preferably about 1-15 ng / ml) insulin-like growth factor (IGF), 1-50 ng / ml (preferably about 1-15 ng / ml) fibroblast growth factor-b (FGF-b), 10−10 to 10−8 M dexamethasone or other appropriate steroid, 0-1 μg / mL linoleic acid, and 10-50 mg / L ascorbic acid. Tissue explant of dental pulp was used to isolate immature DPMSC. The growing culture of DPMSC was maintained under these conditions for 1 day and then the cells were placed, optionally dissociated, in a collagena...

example 1b

Method of Isolating and / or Culturing DPMSC

[0229]Dental pulp was extracted from normal exfoliated human deciduous teeth of 5- to 7-year-old children (12 patients) with informed consent of the donors. Dental pulp was extracted using needle and was transferred into 35-mm Petri dishes (Falcon, BD-Biosciences, Italy) with a proliferation medium composed of F-12 Coon's modified / Ambesi's modified (Gibco) / Medium 199 (Sigma Aldrich, Germany) / CMRL 1066 (Sigma Aldrich, Germany) supplemented with 1.25% of Human serum 1-50 ng / ml platelet-derived growth factor-BB (PDGF-BB, Immunotools, Germany), 1-50 ng / ml epidermal growth factor (EGF, Immunotools, Germany), 1-50 ng / ml insulin-like growth factor-I (IGF-I, Immunotools, Germany), 1-50 ng / ml fibroblast growth factor-I (FGF-b, Immunotools, Germany), 10−10 to 10−8 M dexamethasone (MP), 20-100 μg / L linoleic acid (Sigma, Germany), 10-50 mg / L ascorbic acid (Sigma) and 0.5 ml / L gentamycin (Gibco). The growing culture of DPMSC was maintained under these co...

example 2a

Methods of Phenotype Analysis of DPMSC Cell Population

[0231]Immunofluorescence, Flow-Cytometry, FISH and Immunoblot

[0232]Immunofluorescence and FISH analyses were performed on 4% buffered paraformaldehyde or methanol / acetone fixed cells, while FACS analysis was performed on P3-P5 cells detached from the culture substrate through a short incubation in CTC. Staining was performed either using properly conjugated primary antibodies or with un-conjugated primary antibodies followed by an incubation with conjugated secondary antibodies. Intracellular staining was performed after a permeabilization step utilizing the Intrastain Fixation and Permeabilization kit (Dako, Danmark), following the manufacturer's instructions.

[0233]Image acquisition was carried out by a Confocal Laser Microscope (Leica TCS-SP2, Leica Microsystems, Italy), utilizing either a 63× oil immersion objective (numerical aperture: 1.40) or a 40× oil immersion objective (numerical aperture: 1.25). Epifluorescence and phas...

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PUM

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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.

Description

CROSS-REFERENCES TO RELATED APPLICATIONS[0001]This application is related to and claims the priority benefit of Italian patent application RM2008A000342, filed on Jun. 26, 2008, the entire contents of which are hereby incorporated by reference in its entirety.FIELD OF THE INVENTION[0002]The invention relates generally to compositions and methods for isolating and using dental pulp marrow similar cells (DPMSCs).BACKGROUND OF THE INVENTION[0003]Organ and tissue generation from stem cells and their successive transplantation provide possible treatments for a number of pathologies, making stem cells a central focus of research in many fields. Using stem cells for generation of organs and tissues for transplantation provides a possible therapy for diabetes, Parkinson's disease, liver disease, heart disease, and autoimmune disorders, to name a few. However, there are at least two major problems associated with organ and tissue transplantation. First, there is a shortage of donor organs an...

Claims

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Application Information

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IPC IPC(8): A61K35/12C12N5/07A61P1/18A61P3/10
CPCC12N5/0664C12N2500/36C12N2500/38C12N2501/105C12N2501/39C12N2501/115C12N2501/119C12N2501/135C12N2501/11A61P1/18A61P3/10A61P43/00C12N2500/05C12N2501/33C12N2501/999
Inventor FERRO, FEDERICO
Owner FOUND OF TRANSLATIONAL SCI
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