Optimized multiparametric immunophenotyping of umbilical cord blood cells by flow cytometry

被引:25
作者
Basford, Christina [2 ]
Forraz, Nico [1 ]
McGuckin, Colin [1 ]
机构
[1] CTI LYON, Cell Therapy Res Inst, Lyon, France
[2] Newcastle Univ, Inst Human Genet, Stem Cell Grp, Newcastle Upon Tyne, Tyne & Wear, England
关键词
ISOTYPE CONTROLS; STEM-CELLS; TRANSPLANTATION; LYMPHOCYTES; TIME;
D O I
10.1038/nprot.2010.88
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Umbilical cord blood is a key source of stem cells for transplantation and regenerative medicine. To maximize each cord blood sample, it is important to analyze its cellular populations. Many cord blood banks focus on counts of total nucleated cells and/or cells that carry the CD34 antigen, but this limited focus does not give a true estimation of cord blood content, quality and appropriateness for use. This protocol is the first of its kind to enable a comprehensive investigation of cord blood cellular populations. Using multicolor flow cytometry it is possible to examine expression of 26 antigens-including hematopoietic markers CD45 and CD34, immune markers CD19 and CD3, and HLA-using a total of only 1 x 10(6) cells from each unit for both pre- and post-processing. The samples are stained, lysed, washed and analyzed flow cytometrically. This method also provides valuable information beyond cord blood composition and quality; for example, it can also be used to assess whether maternal factors affect CD34(+) cell numbers. Collection, processing, cryopreservation and initial flow cytometry take similar to 5 h.
引用
收藏
页码:1337 / 1346
页数:10
相关论文
共 28 条
[1]   Human mesenchymal stem cells form Purkinje fibers in fetal sheep heart [J].
Airey, JA ;
Almeida-Porada, G ;
Colletti, EJ ;
Porada, CD ;
Chamberlain, J ;
Movsesian, M ;
Sutko, JL ;
Zanjani, ED .
CIRCULATION, 2004, 109 (11) :1401-1407
[2]  
Alamo AL, 2000, CYTOMETRY, V42, P363, DOI 10.1002/1097-0320(20001215)42:6<363::AID-CYTO1003>3.0.CO
[3]  
2-E
[4]   Umbilical cord blood processing using Prepacyte-CB increases haematopoietic progenitor cell availability over conventional Hetastarch separation [J].
Basford, C. ;
Forraz, N. ;
Habibollah, S. ;
Hanger, K. ;
McGuckin, C. P. .
CELL PROLIFERATION, 2009, 42 (06) :751-761
[5]   Fluorescence-intensity multiplexing: Simultaneous seven-marker, two-color immunophenotyping using flow cytometry [J].
Bradford, JA ;
Buller, G ;
Suter, M ;
Ignatius, M ;
Beechem, JM .
CYTOMETRY PART A, 2004, 61A (02) :142-152
[6]   Umbilical cord blood transplantation - how, when and for whom? [J].
Cohen, Y ;
Nagler, A .
BLOOD REVIEWS, 2004, 18 (03) :167-179
[7]  
D'Arena G, 1998, HAEMATOLOGICA, V83, P197
[8]   Transplantation of umbilical-cord blood in babies with infantile Krabbe's disease [J].
Escolar, ML ;
Poe, MD ;
Provenzale, JM ;
Richards, KC ;
Allison, J ;
Wood, S ;
Wenger, DA ;
Pietryga, D ;
Wall, D ;
Champagne, M ;
Morse, R ;
Krivit, W ;
Kurtzberg, J .
NEW ENGLAND JOURNAL OF MEDICINE, 2005, 352 (20) :2069-2081
[9]   Human umbilical cord blood banking and transplantation: a state of the art [J].
Fasouliotis, SJ ;
Schenker, JG .
EUROPEAN JOURNAL OF OBSTETRICS & GYNECOLOGY AND REPRODUCTIVE BIOLOGY, 2000, 90 (01) :13-25
[10]   HEMATOPOIETIC RECONSTITUTION IN A PATIENT WITH FANCONIS ANEMIA BY MEANS OF UMBILICAL-CORD BLOOD FROM AN HLA-IDENTICAL SIBLING [J].
GLUCKMAN, E ;
BROXMEYER, HE ;
AUERBACH, AD ;
FRIEDMAN, HS ;
DOUGLAS, GW ;
DEVERGIE, A ;
ESPEROU, H ;
THIERRY, D ;
SOCIE, G ;
LEHN, P ;
COOPER, S ;
ENGLISH, D ;
KURTZBERG, J ;
BARD, J ;
BOYSE, EA .
NEW ENGLAND JOURNAL OF MEDICINE, 1989, 321 (17) :1174-1178