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(0.01-0.03%)

Basement membrane Extracellular matrix and

CytokinesCell-cell interactions

Niche , Selfrenew and Diffrentation relationship

How we can identify Stem cells

1. In vitro single cell colony assay

2. Single cell PCR , looking for upregulation and downregulation of lineage related genes

3. 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 weeksOP9DL1 cell line

HSC Transplantation assay

Blood, 2009, 113:3453-60

Hematopoiesis - Blood Cell Formation

Myeloid Lymphoid

ErythrocytePlatelet

MacrophageGranulocyte

NK cell B cell T cellSelf-renew

al

Stem Cells

Committed progenitors

Mature cells

Bone

mar

row

Thymus,Blood,Spleen

CD34-Flt3-CD150+Lin-ScahighKithigh

CD34+Flt3- CD150-Lin-ScahighKithigh

LT-HSC

ST-HSC

MPPs CD34+Flt3+Lin-ScahighKithigh

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 CD1332. Express low level of CD24 and PNA(peanut agglotinin in SVZ mic3. 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

Thank you so much for your attention