FAQ
- 用途別細胞培養
- ALDEFLUORをヒトやマウス以外の細胞に対するアッセイで使用した例はありますか？
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STEMCELL Technologies社では試したことはないものの、ALDEFLUORの開発者らがイヌ、ブタ、アカゲザル細胞に対して検討した例が学会発表にて紹介されています。以下、要旨を紹介致します。
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Identification of Aldehyde Dehydrogenase (ALDH) Bright Cells in Monkey, Dog and Pig.
M. N. Fiordalisi1, A. Bonifacino2, R. de Silva3, R. E. Donahue2, H. R. Phillips4, N. R. Haley1, A. E. Balber
(1: 1STEMCO Biomedical, Durham, NC, 2: Hematology Branch, National Heart Lung and Blood Institute, NIH, Bethesda, MD, 3: Cardiovascular Branch, National Heart Lung and Blood Institute, NIH, Bethesda, MD, 4: Duke University, Durham, NC. ISCT 2005.)
A variety of human stem and progenitor cells express high levels of aldehyde dehydrogenase. These cells can be identified using ALDEFLUOR® a commercially available flow cytometric ALDH assay. Human cell populations sorted from bone marrow (BM) on the basis of high ALDH expression and low side scatter (ALDHbrSSClo) are highly active in culture assays indicative of angiogenic potential. We now report that ALDHbrSSClo cells can be detected in mobilized peripheral blood (mPBSC) of rhesus macaques and in BM of dogs and swine. These species are widely used in preclinical models of cardiovascular disease. Flow sorting ALDHbrSSClo cells could provide an alternative method to antibody-based cell selection for identifying cells with regenerative capacity. We used the ALDEFLUOR® kit as recommended except that 0.1% NaN3 and 50 μM verapamil was added to all buffers and samples were maintained on ice. Rhesus macaque ALDHbrSSClo cells represented 0.1-0.2% of nucleated mPBSC were positive for CD34 (98%) and negative for CD2, CD14 and CD21. ALDHbrSSClo cells comprised 63% of the CD34+ population. No ALDHbrSSClo cells were detected prior to G-CSF mobilization. We used higher ALDEFLUOR® substrate (4.8 μM) for optimal detection of ALDHbrSSClo cells in beagle and swine. ALDHbrSSClo cells represented 0.45-0.65% of total mononuclear canine BM cells and did not react with antibodies to canine CD5, CD8, CD14 B cells or neutrophils. Resolution of ALDHbrSSClo cells was enhanced following immunomagnetic depletion with this antibody cocktail. The canine BM ALDHbrSSClo population showed little reaction (0.5-3%) with anti-canine CD34 antibody (clone 1H6). We identified a population (2%) of ALDHbrSSClo cells in BM from Yucatan mini or Yorkshire swine after partial depletion with CD5, CD21 and SWC3a (reactive against monocytes/granulocytes). Identification of ALDHbrSSClo cells in swine BM required lineage depletion. Flow sorting of ALDHbrSSClo cells permits investigation of these cells for mediating cardiac repair in various species.
