stem cell identification nozad h. stem cell workshop stem cell research center tabriz medical...
Post on 19-Jan-2016
218 Views
Preview:
TRANSCRIPT
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
top related