LW_BCH4122_Lecture3_2009

LW_BCH4122_Lecture3_2009 - Lecture 3. Surrogate markers to...

Info iconThis preview shows pages 1–12. Sign up to view the full content.

View Full Document Right Arrow Icon
Lecture 3. Surrogate markers to characterize adult stem cells Lisheng WANG Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
How are adult stem cells characterized? The basic characterization of an adult stem cell are a single cell (clonal) that self-renews and generates differentiated cells. Clonal assay will remove doubts about contamination with other cell types. The most rigorous assessment of these characteristics is to prospectively purify a population of cells ( usually based on cell surface macromolecules ) , transplant a single cell into an acceptable host without any intervening in vitro culture, and observe self-renewal and tissue, organ, or lineage reconstitution. However, this type of in vivo reconstitution assay is not well defined for many types of adult stem cells because of technical difficulties.
Background image of page 2
Many different types of stem cells are found in very small populations in the human body (for instance 1 in 100,000 cells in blood). Stem cells look just like any other cells under a microscope in the tissue where they reside. Identification of these rare type of cells (a process like to find a needle in a haystack) is based on stem cell "markers." These markers are macromolecules located either inside stem cells or on cell surface. Use of macromolecules to identify and isolate adult stem cells
Background image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Characterize adult stem cell based on surface markers In vivo and in vitro analyses of single adult stem cells Select and purify macromolecules that mark adult stem cells Generate specific antibodies against a given specific macromolecules Label the antibodies with fluorescent probe/enzyme/biotin Purify adult stem cells according to the expression of specific macromolecules that are recognized by specific antibodies
Background image of page 4
A given macromolecule can be recognized by a specific antibody generated from an immunized animal Antibody IgG Source of image: http://www.accessexcellence.org/RC/VL/GG/antiBD_mol.html
Background image of page 5

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
The specific antibody will be conjugated with a fluorescent probe, forming a specific fluorescent tag Image source : http://www.piercenet.com/Objects/view.cfm?type=Page&ID=412BAC2E-F0BC-4ABA-A79B-887E413910AB
Background image of page 6
r ght The fluorescent probe can be detected under UV light FITC Image source: http://www. hmds.org.uk/figures/mabs.jpg Image source: http://immuneweb.xxmc.edu.cn/monoclonal/fluor-ab.html
Background image of page 7

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
A protein expressed on the membrane of an adult stem cell will be recognized by a specific antibody conjugated with fluorescent probe (for example FITC). Image source: http://stemcells.nih.gov/StaticResources/info/scireport/images/figuree1.jpg
Background image of page 8
Intra-cellular macromolecule can be stained together with a cell surface macromolecule
Background image of page 9

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Observation of the cell markers (macromolecules) under fluorescent microscope
Background image of page 10
Image source: http://stemcells.nih.gov/StaticResources/info/scireport/images/figuree2.jpg
Background image of page 11

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Image of page 12
This is the end of the preview. Sign up to access the rest of the document.

This note was uploaded on 12/11/2009 for the course BCH bch4122 taught by Professor Wang during the Spring '09 term at University of Ottawa.

Page1 / 37

LW_BCH4122_Lecture3_2009 - Lecture 3. Surrogate markers to...

This preview shows document pages 1 - 12. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online