Item request has been placed! ×
Item request cannot be made. ×
loading  Processing Request

Comparison of Three CD3-Specific Separation Methods Leading to Labeled and Label-Free T Cells.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
اقرأ على الانترنت اقرأ أكثر حفظ في قائمتي
  • معلومة اضافية
    • المصدر:
      Publisher: MDPI Country of Publication: Switzerland NLM ID: 101600052 Publication Model: Electronic Cited Medium: Internet ISSN: 2073-4409 (Electronic) Linking ISSN: 20734409 NLM ISO Abbreviation: Cells Subsets: MEDLINE
    • بيانات النشر:
      Original Publication: Basel, Switzerland : MDPI
    • الموضوع:
      Staining and Labeling*; CD3 Complex/*metabolism ; Cell Separation/*methods ; T-Lymphocytes/*cytology; Cell Count ; Cell Proliferation ; Cell Shape ; Cell Survival ; Cytokines/biosynthesis ; Erythrocytes/cytology ; Humans
    • نبذة مختصرة :
      T cells are an essential part of the immune system. They determine the specificity of the immune response to foreign substances and, thus, help to protect the body from infections and cancer. Recently, T cells have gained much attention as promising tools in adoptive T cell transfer for cancer treatment. However, it is crucial not only for medical purposes but also for research to obtain T cells in large quantities, of high purity and functionality. To fulfill these criteria, efficient and robust isolation methods are needed. We used three different isolation methods to separate CD3-specific T cells from leukocyte concentrates (buffy coats) and Ficoll purified PBMCs. To catch the target cells, the Traceless Affinity Cell Selection (TACS ® ) method, based on immune affinity chromatography, uses CD-specific low affinity Fab-fragments; while the classical Magnetic Activated Cell Sorting (MACS ® ) method relies on magnetic beads coated with specific high affinity monoclonal antibodies. The REAlease ® system also works with magnetic beads but, in contrast to MACS ® , low-affinity antibody fragments are used. The target cells separated by TACS ® and REAlease ® are "label-free", while cells isolated by MACS ® still carry the cell specific label. The time required to isolate T cells from buffy coat by TACS ® and MACS ® amounted to 90 min and 50 min, respectively, while it took 150 min to isolate T cells from PBMCs by TACS ® and 110 min by REAlease ® . All methods used are well suited to obtain T cells in large quantities of high viability (>92%) and purity (>98%). Only the median CD4:CD8 ratio of approximately 6.8 after REAlease ® separation differed greatly from the physiological conditions. MACS ® separation was found to induce proliferation and cytokine secretion. However, independent of the isolation methods used, stimulation of T cells by anti CD3/CD28 resulted in similar rates of proliferation and cytokine production, verifying the functional activity of the isolated cells.
    • References:
      Nature. 1983 Jun 30;303(5920):808-10. (PMID: 6191218)
      PLoS One. 2013 Sep 06;8(9):e74745. (PMID: 24040334)
      Channels (Austin). 2021 Dec;15(1):53-66. (PMID: 33356811)
      Cytometry A. 2019 Feb;95(2):234-241. (PMID: 30378734)
      Rep Prog Phys. 2015 Jan;78(1):016601. (PMID: 25471081)
      Nano Lett. 2013;13(11):5619-26. (PMID: 24117051)
      Sci Rep. 2018 Nov 13;8(1):16731. (PMID: 30425259)
      Br J Haematol. 2015 Jan;168(1):26-37. (PMID: 25145701)
      Cytometry. 1996 Sep 15;26(3):231-2. (PMID: 8889397)
      Nat Med. 2018 Oct;24(10):1499-1503. (PMID: 30275568)
      Clin Exp Immunol. 1987 Apr;68(1):146-55. (PMID: 3115639)
      Cytometry. 1990;11(2):231-8. (PMID: 1690625)
      Leukemia. 2012 Mar;26(3):381-9. (PMID: 21886171)
      Cold Spring Harb Perspect Biol. 2010 Apr;2(4):a005140. (PMID: 20452950)
      Leukemia. 2011 Mar;25(3):498-505. (PMID: 21135860)
      Cytometry A. 2010 Jul;77(7):643-7. (PMID: 20583279)
      Leukemia. 2016 Feb;30(2):492-500. (PMID: 26369987)
      Immunity. 2007 Sep;27(3):393-405. (PMID: 17892848)
      Oncoimmunology. 2015 May 5;4(11):e1046673. (PMID: 26451319)
      Cytokine. 2015 Jul;74(1):5-17. (PMID: 25458968)
      J Clin Invest. 2016 Jun 1;126(6):2123-38. (PMID: 27111235)
      Cell Med. 2019 Jan 22;11:2155179018822781. (PMID: 32634192)
      Int J Mol Sci. 2019 Dec 10;20(24):. (PMID: 31835562)
      Nat Med. 1995 Dec;1(12):1279-83. (PMID: 7489409)
      Nat Rev Drug Discov. 2021 Mar;20(3):166. (PMID: 33574586)
      Immunity. 2018 Feb 20;48(2):202-213. (PMID: 29466753)
      Korean J Hematol. 2011 Mar;46(1):18-23. (PMID: 21461299)
      Oncol Rep. 2019 Jun;41(6):3455-3463. (PMID: 30942469)
      Proc Natl Acad Sci U S A. 2019 Mar 26;116(13):5914-5919. (PMID: 30850527)
    • Grant Information:
      100362591 Development Bank of Saxony
    • Contributed Indexing:
      Keywords: CD3; MACS®; REAlease®; T cells; TACS®; isolation; proliferation; separation
    • الرقم المعرف:
      0 (CD3 Complex)
      0 (Cytokines)
    • الموضوع:
      Date Created: 20211127 Date Completed: 20211227 Latest Revision: 20211227
    • الموضوع:
      20240829
    • الرقم المعرف:
      PMC8616525
    • الرقم المعرف:
      10.3390/cells10112824
    • الرقم المعرف:
      34831046