[Detection and analysis of CD271, CD133 and CD34 expressions in bone marrow cells by flow cytometry with three color fluorescence labelling].
This study was purposed to detect the expressions of CD271, CD133 and CD34, and to analyze the correlation of CD271 with CD133 and CD133 with CD34 expressions. The human bone marrow cells (BMCs) and mononucleated cells (MNCs) were detected by flow cytometry with CD45-PerCP, CD271-FITC, CD133-PE and CD34-FITC labelling according to different combinations of design, cells were located and selected repeatedly by FSC, SSC and CD45 after acquirement, then the expressions of CD271, CD133 and CD34 were detected by flow cytometry. The results showed that the expressions of CD271, CD133 and CD34 in BMCs were 0.16%, 0.20% and 0.43% respectively, while their expressions were 0.49%, 0.47% and 1.07% respectively after isolation of MNCs. The co-expressions of CD271(+)CD133(+) before and after isolation of MNCs were (0.02 +/- 0.01)% and (0.03 +/- 0.02)% respectively. The co-expression of CD133(+) and CD34(+) before and after isolation of MNCs were (0.18 +/- 0.11)% and (0.42 +/- 0.23)% respectively (p < 0.01); meanwhile about 90% of cells with CD133(+) expressed CD34 and 40% of cells with CD34(+) expressed CD133. It is concluded that the established method of detection using flow cytometry with three color fluorescence labelling can be used to detect expression of CD271, CD133 and CD34 in BMCs. The cells with CD271 are different from cells with CD133 and CD34, which suggests that the CD271 may be of important role in evaluating and guiding the clinical application of BM MSCs.
Zhou
Jun
J
Department of Blood Transfusion, General Hospital, Beijing Military Area, Beijing 100700, China. zj990222@tom.com
Zhu
Bing
B
DU
Hai-Yan
HY
Sun
Tian-Sheng
TS
Zhang
Chang-Hong
CH
Yang
Lian-Gui
LG
D000071916:AC133 Antigen; D015703:Antigens, CD / Q000378:metabolism*; D018952:Antigens, CD34 / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D002460:Cell Line; D005434:Flow Cytometry; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D017475:Receptors, Nerve Growth Factor / Q000378:metabolism*
D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; D006023:Glycoproteins; C484065:NGFR protein, human; D009419:Nerve Tissue Proteins; C000606023:PROM1 protein, human; D010455:Peptides; D017475:Receptors, Nerve Growth Factor