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

Stem Cell Characteristics 1.‘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( %)

Basement membrane Extracellular matrix and Cytokines Cell-cell interactions

Niche, Selfrenew and Diffrentation relationship

How we can identify Stem cells 1.In vitro single cell colony assay 1.Single cell PCR, looking for upregulation and downregulation of lineage related genes 1.In Vivo assay( Stem cell Transplantation)

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

HSC Transplantation assay Blood, 2009, 113:

Hematopoiesis - Blood Cell Formation Myeloid Lymphoid Erythrocyte Platelet Macrophage Granulocyte NK cellB cellT cell Self-renewal Stem Cells Committed progenitors Mature cells Bone marrow Thymus, Blood, Spleen CD34 - Flt3 - CD150 + Lin - Sca high Kit high CD34 + Flt3 - CD150 - Lin - Sca high Kit high LT-HSC ST-HSC MPPsCD34 + Flt3 + Lin - Sca high Kit high

Flow cytometry analysis of HSC

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 haematopoietic and endothelial cell markers

Identifying the mesenchimal stem cell

Neural Stem cell 1. Express CD 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

Neural stem cell Nestin, Sox2

Stage Specific Embryonic markers

Embryonic stem cell markers

stem cell in Intestine crypt

Colonic crypt organization, patterns of stem cell divisions

Intestine stem cell markers

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

Cancer stem cell

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