What is CAFC assay???

Discussion of all aspects of cellular structure, physiology and communication.

Moderators: honeev, Leonid, amiradm, BioTeam

Post Reply
User avatar
[email protected]
Garter
Garter
Posts: 6
Joined: Sun Aug 07, 2005 9:52 am

What is CAFC assay???

Post by [email protected] » Sun Aug 07, 2005 9:54 am

Does anyone know what is CAFC (cobblestone area forming cell) assay? What is it ususally used for?

User avatar
ginny
Garter
Garter
Posts: 19
Joined: Wed Jul 13, 2005 7:39 pm
Location: Los Angeles
Contact:

hope i helped you !!!!!!!!!!!!!!!!!

Post by ginny » Tue Aug 09, 2005 6:40 pm

Cobblestone Area-Forming Cells in Human Cord Blood Are Heterogeneous and Differ from Long-Term Culture-Initiating Cells.

Formed in normal bone marrow stroma.

The characterization and quantitation of primitive human hematopoietic cells is essential for understanding the developmental sequence of hematopoiesis, stem cell manipulation, and transplantation. Assay of primitive hematopoietic cells is difficult because of their rarity and expression of surface markers such as CD38 and Thy-1 [1], with different subsets representing stages of differentiation. The most primitive hematopoietic cells can ultimately only be recognized by their ability to repopulate marrow of a myeloablated recipient of the same species. However, a physiological in vitro approach is to measure long-term culture-initiating cells (LTC-IC) [2] in a modified Dexter culture system [3] using the fact that after 5 or more weeks of culture, LTC-IC will produce de novo colony-forming cells (CFC).

Alternatively, the cobblestone area-forming cell (CAFC) readout exploits a visual end point for the assay, eliminating trypsinization and the CFC assay. Wells are scored as being positive or negative for the presence of cobblestone areas (CA, tightly knit group of phase-dark, angular cells in the stroma) and can be scored at more than one time point, e.g., after 5 and 8 weeks. In the murine model, CAFCs seen between days 28–45 of long-term culture are directly related to bone marrow repopulating ability [4].

Both LTC-IC and CAFC frequencies have been reported in the study of stem cell storage [5], expansion [6, 7], or mobilization [8]. In the clinical setting these assays have been used to assess stem cell defects (e.g., in aplastic anemia [9] and Diamond Blackfan anemia [10]) and to investigate bone marrow reserve after stem cell transplantation [11, 12] or in patients with autoimmune cytopenias [13].

In the present work, we set out to measure the number of primitive cells in cord blood (CB) that had been processed for clinical banking. We chose limiting dilution on cryopreserved human bone marrow stroma feeder layers [14] with the visual CAFC readout. It soon became apparent that further validation of the assay was required. We have therefore compared the frequency of CAFC with LTC-IC in CD34-enriched and CD34-depleted fractions of CB.


byeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee :roll:
I would like to become a pediactirc doctor. I also want to get into med-school. I attend UCLA.

User avatar
[email protected]
Garter
Garter
Posts: 6
Joined: Sun Aug 07, 2005 9:52 am

Thanks!!

Post by [email protected] » Thu Aug 11, 2005 12:51 pm

Thanks for ur useful information!! I am currently doing my degree on Biological Sciences!! And our lecturer loves to give us extra assignments based on the recent pulished scientific articles!! :cry:

peasnquiet
Garter
Garter
Posts: 1
Joined: Thu Sep 08, 2005 8:57 pm

Post by peasnquiet » Thu Sep 08, 2005 8:59 pm

Oh crud.

Huh. Same school. And when I typed in CAFC assay on google, guess what was the first search result that came up?

*rolls eyes*

app1eg
Garter
Garter
Posts: 1
Joined: Thu Aug 25, 2005 5:12 pm

Post by app1eg » Fri Sep 09, 2005 1:25 am

lol
im from sbs in ntu too..
gawd.. the world is just so small.

Post Reply

Who is online

Users browsing this forum: No registered users and 10 guests