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Stem Cell Identification

In The Name of God. Stem Cell Identification. Nozad H. Stem cell workshop Stem cell Research center Tabriz medical university. Stem Cell Characteristics. ‘Blank cells’ ( unspecialized ) 2.Capable of dividing and renewing themselves for long periods of time

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Stem Cell Identification

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  1. In The Name of God Stem Cell Identification Nozad H. Stem cell workshop Stem cell Research center Tabriz medical university

  2. Stem Cell Characteristics ‘Blank cells’ (unspecialized) 2.Capable of dividing and renewing themselves for long periods of time (proliferation and self renewal) 3. Have the potential to give rise to specialized cell types (differentiation) 4. Plasticity and Cell based therapies 5. Very low percentage available(0.01-0.03%)

  3. Extracellular matrix and Basement membrane Cell-cell interactions Cytokines

  4. Niche , Selfrenew and Diffrentation relationship

  5. How we can identify Stem cells • In vitro single cell colony assay • Single cell PCR , looking for upregulation and downregulation of lineage related genes • In Vivo assay( Stem cell Transplantation)

  6. Sorting single cell stem cell to evaluate the lineage potential Single stem cell OP9 cell line Analysis Co-cultures OP9DL1 cell line For 3 to 5 weeks

  7. HSC Transplantation assay Blood, 2009, 113:3453-60

  8. Hematopoiesis - Blood Cell Formation Self-renewal LT-HSC CD34-Flt3-CD150+Lin-ScahighKithigh Stem Cells ST-HSC CD34+Flt3- CD150-Lin-ScahighKithigh MPPs CD34+Flt3+Lin-ScahighKithigh Bone marrow Lymphoid Myeloid Committed progenitors Mature cells Thymus, Blood, Spleen Erythrocyte Macrophage NK cell B cell T cell Platelet Granulocyte

  9. Flow cytometry analysis of HSC

  10. Identifying mesenchymal stem cell 1. A heterogeneous population, mixed population of fibroblast-like plastic-adherent cells 2. differentiate into a range of mature connective tissue cell types . 3. MSCs have been isolated from skeletal muscle, adipose tissue, deciduous teeth, 4. No surface antigen individually, or in combination, has been unified as a “supreme” marker 5. A supportive role to HSCs during hematopoiesis. 6. expressed markers on MSC depending of the source of MSCs 7. MSC do not express haematopoieticand endothelial cell markers

  11. Identifying the mesenchimal stem cell

  12. Neural Stem cell 1. Express CD133 2. Express low level of CD24 and PNA(peanut agglotinin in SVZ mic 3. CXCR4, FGF R4 , BMI-1, Brg1, FABP7, Nestin 3. Combination of this marker by Nestin gene expression is the best NSCs

  13. Neural stem cell Nestin, Sox2

  14. Stage Specific Embryonic markers

  15. Embryonic stem cell markers

  16. stem cell in Intestine crypt

  17. Colonic crypt organization, patterns of stem cell divisions

  18. Intestine stem cell markers

  19. Inducing a "forced" expression of specific genes(Oct3/4, Sox2, Klf4 and cMyc) in somatic cells

  20. Cancer stem cell

  21. Thank you so much for your attention

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