PMID- 30426739
OWN - NLM
STAT- MEDLINE
DCOM- 20190516
LR  - 20190516
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 90
IP  - 24
DP  - 2018 Dec 18
TI  - Multiplexed Label-Free Fractionation of Peripheral Blood Mononuclear Cells for 
      Identification of Monocyte-Platelet Aggregates.
PG  - 14535-14542
LID - 10.1021/acs.analchem.8b04415 [doi]
AB  - Monocytes and platelets play key roles in atherosclerosis and thrombosis, and 
      circulating monocyte-platelet aggregates (MPA) in blood have been widely proposed 
      as surrogate biomarkers for cardiovascular risk stratification and monitoring 
      antiplatelet therapies. However, conventional MPA characterization is based on 
      whole-blood fixation and flow cytometry analysis which adversely affect cell 
      viability and detection accuracy due to significant leukocyte and platelet 
      contaminations. Herein, we introduce a rapid and label-free microfluidic approach 
      for complete size-based fractionation of peripheral blood mononuclear cells 
      (PBMCs) into monocytes, lymphocytes, and platelets. The developed biochip enables 
      gentle sorting of intact MPA in the enriched monocytes with efficient depletion 
      of lymphocytes and platelets for accurate MPA quantification. We first compared 
      the developed microfluidic technology (dean flow fractionation, DFF) with 
      standard magnetic negative isolation (MACS) and observed that DFF-sorted 
      monocytes had similar viability, purity, and key immune functions (phagocytosis, 
      macrophage differentiation) as MACS-sorted monocytes. As proof of concept for 
      diabetes testing, we isolated and characterized monocytes/MPA from a cohort of 
      healthy ( n = 5) and type 2 diabetes mellitus (T2DM) subjects ( n = 8) in PBMCs 
      and DFF-sorted monocytes. High-speed imaging, immunofluorescence, and flow 
      cytometry analysis clearly indicated higher levels of MPA in T2DM patients ( P < 
      0.05) with enhanced MPA detection sensitivity in DFF-sorted monocytes ( P < 
      0.05). Taken together, the developed DFF technology greatly facilitates 
      high-throughput ( approximately 130 muL min(-1)) label-free isolation of monocyte/MPA from PBMCs 
      and can be further developed into a clinical tool for point-of-care 
      cardiovascular risk stratification in metabolic disorders including T2DM.
FAU - Tay, Hui Min
AU  - Tay HM
AUID- ORCID: 0000-0002-4981-5994
AD  - School of Mechanical and Aerospace Engineering , Nanyang Technological University 
      , 50 Nanyang Avenue , 639798 , Singapore.
FAU - Yeap, Wei Hseun
AU  - Yeap WH
AD  - Singapore Immunology Network , Agency for Science, Technology and Research , 8a 
      Biomedical Grove , 138648 , Singapore.
FAU - Dalan, Rinkoo
AU  - Dalan R
AD  - Endocrine and Diabetes , Tan Tock Seng Hospital , 11 Jalan Tan Tock Seng , 308433 
      , Singapore.
FAU - Wong, Siew Cheng
AU  - Wong SC
AD  - Singapore Immunology Network , Agency for Science, Technology and Research , 8a 
      Biomedical Grove , 138648 , Singapore.
AD  - School of Biological Sciences , Nanyang Technological University , 60 Nanyang 
      Drive , 637551 , Singapore.
FAU - Hou, Han Wei
AU  - Hou HW
AD  - School of Mechanical and Aerospace Engineering , Nanyang Technological University 
      , 50 Nanyang Avenue , 639798 , Singapore.
AD  - Lee Kong Chian School of Medicine , Nanyang Technological University , Clinical 
      Sciences Building, 11 Mandalay Road , 308232 , Singapore.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20181127
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Cytokines)
SB  - IM
MH  - Blood Platelets/*cytology/metabolism
MH  - Case-Control Studies
MH  - Cell Differentiation
MH  - Cytokines/metabolism
MH  - Diabetes Mellitus, Type 2/metabolism/pathology
MH  - Flow Cytometry
MH  - Humans
MH  - Leukocytes, Mononuclear/cytology
MH  - Microfluidics/*methods
MH  - Microscopy, Fluorescence
MH  - Monocytes/*cytology/metabolism
MH  - Phagocytosis
EDAT- 2018/11/15 06:00
MHDA- 2019/05/17 06:00
CRDT- 2018/11/15 06:00
PHST- 2018/11/15 06:00 [pubmed]
PHST- 2019/05/17 06:00 [medline]
PHST- 2018/11/15 06:00 [entrez]
AID - 10.1021/acs.analchem.8b04415 [doi]
PST - ppublish
SO  - Anal Chem. 2018 Dec 18;90(24):14535-14542. doi: 10.1021/acs.analchem.8b04415. 
      Epub 2018 Nov 27.