PMID- 29434528
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20220311
IS  - 1480-9222 (Print)
IS  - 1480-9222 (Electronic)
IS  - 1480-9222 (Linking)
VI  - 20
DP  - 2018
TI  - Antitumor Efficacy of Human Monocyte-Derived Dendritic Cells: Comparing Effects 
      of two Monocyte Isolation Methods.
PG  - 4
LID - 10.1186/s12575-018-0069-6 [doi]
LID - 4
AB  - BACKGROUND: Dendritic cells (DCs), which can be used as anti-cancer vaccines, are 
      generally obtained in vitro from isolated CD14(+) monocytes (MoDCs). This 
      generates high cell numbers and allows instructing DCs to guarantee effective 
      antitumor responses. However, the impact of the monocyte isolation step in the 
      antitumor effectiveness of the generated MoDCs is still unknown. Here, we 
      compared the most used immunomagnetic technologies for monocyte isolation: 
      magnetic activated cell sorting (MACS) from Miltenyi Biotec and EasySep from STEM 
      CELL. RESULTS: MACS technology allowed a higher monocyte yield and purity and, by 
      flow cytometry, monocytes displayed higher size and lower granularity. In the 
      resting state, EasySep_MoDCs showed a higher basal expression of HLA-DR, and no 
      significant response to stimulation by LPS and TNF-alpha. When stimulated with whole 
      tumor cells lysates, both MoDCs expressed similar levels of maturation and 
      co-stimulatory markers. However, when cultured with autologous T cells, 
      MACS_MoDCs induced significantly higher IFN-gamma secretion than EasySep_MoDCs, 
      indicating a stronger induction of Th1 cell response profile. Concordantly, T 
      cells induced by MACS_MoDCs also showed a higher release of cytotoxic granules 
      when in contact with tumor cells. CONCLUSIONS: Overall, both the MACS and the 
      EasySep isolation immunomagnetic technologies provide monocytes that 
      differentiate into viable and functional MoDCs. In our experimental settings, 
      resting EasySep_MoDCs showed a higher basal level of maturation but show less 
      responsivity to stimuli. On the other hand, MACS_MoDCs, when stimulated with 
      tumor antigens, showed better ability to stimulate Th1 responses and to induce T 
      cell cytotoxicity against tumor cells. Thus, monocyte isolation techniques 
      crucially affect MoDCs' function and, therefore, should be carefully selected to 
      obtain the desired functionality.
FAU - Marques, Graca S
AU  - Marques GS
AD  - 1CEDOC, NOVA Medical School/Faculdade de Ciencias Medicas, Universidade NOVA de 
      Lisboa, Lisbon, Portugal. ISNI: 0000000121511713. GRID: grid.10772.33
FAU - Silva, Zelia
AU  - Silva Z
AD  - 1CEDOC, NOVA Medical School/Faculdade de Ciencias Medicas, Universidade NOVA de 
      Lisboa, Lisbon, Portugal. ISNI: 0000000121511713. GRID: grid.10772.33
AD  - 2UCIBIO, Departamento Ciencias da Vida, Faculdade de Ciencias e Tecnologia, 
      Universidade NOVA de Lisboa, Lisbon, Portugal. ISNI: 0000000121511713. GRID: 
      grid.10772.33
FAU - Videira, Paula A
AU  - Videira PA
AD  - 1CEDOC, NOVA Medical School/Faculdade de Ciencias Medicas, Universidade NOVA de 
      Lisboa, Lisbon, Portugal. ISNI: 0000000121511713. GRID: grid.10772.33
AD  - 2UCIBIO, Departamento Ciencias da Vida, Faculdade de Ciencias e Tecnologia, 
      Universidade NOVA de Lisboa, Lisbon, Portugal. ISNI: 0000000121511713. GRID: 
      grid.10772.33
AD  - 3CDG & Allies - Professionals and Patient Associations International Network (CDG 
      & Allies - PPAIN), Faculdade de Ciencias e Tecnologia, Universidade NOVA de 
      Lisboa, Lisbon, Portugal. ISNI: 0000000121511713. GRID: grid.10772.33
LA  - eng
PT  - Journal Article
DEP - 20180202
PL  - England
TA  - Biol Proced Online
JT  - Biological procedures online
JID - 100963717
PMC - PMC5796591
OTO - NOTNLM
OT  - Cancer vaccines
OT  - Immunomagnetic cell isolation
OT  - Monocyte-derived dendritic cells (MoDCs)
OT  - T-cell cytotoxicity
COIS- This article does not contain any studies with human participants. The approval 
      to use peripheral blood mononuclear cells (PBMCs), from leuko-platelet 
      concentrates from healthy anonymous donors, was obtained from the Portuguese 
      Blood and Transplantation Institute (Instituto Portugues do Sangue e da 
      Transplantacao - IPST) (P-0114/CD/12HT/pt); after the approval from the 
      institutional ethical committee. Permission to use the blood cell donations is 
      obtained after informed consent from anonymous donors.Not applicableThe authors 
      declare that they have no competing interests.Springer Nature remains neutral 
      with regard to jurisdictional claims in published maps and institutional 
      affiliations.
EDAT- 2018/02/13 06:00
MHDA- 2018/02/13 06:01
PMCR- 2018/02/02
CRDT- 2018/02/14 06:00
PHST- 2017/11/16 00:00 [received]
PHST- 2018/01/17 00:00 [accepted]
PHST- 2018/02/14 06:00 [entrez]
PHST- 2018/02/13 06:00 [pubmed]
PHST- 2018/02/13 06:01 [medline]
PHST- 2018/02/02 00:00 [pmc-release]
AID - 69 [pii]
AID - 10.1186/s12575-018-0069-6 [doi]
PST - epublish
SO  - Biol Proced Online. 2018 Feb 2;20:4. doi: 10.1186/s12575-018-0069-6. eCollection 
      2018.