Immunotherapy Workflow - Step 1. Cell Collection & Isolation

Medical professionals collect blood containing immune cells from patient. Specialized equipment isolates the immune cells from the rest of the blood.
T cells can then be enriched and washed to separate them from other cells.

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cell collection & isolation

​Your challenges

  • ​Virus or Pathogen manipulations in safe conditions?
  • PBMC with RBC contamination, debris, low recovery, poor viability, … How do you increase quality of PBMC or cell preparation ?
  • Is your cell count and viability analysis method easy and reliable?
  • Do your cryopreservation process impact T cell recovery, viability, or functionality?

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Understanding Centrifugal Elutriation's Vital Role in Advanced Flow Cytometry Labs

Welcome to a comprehensive guide on centrifugal elutriation's pivotal role in modernizing and optimizing flow cytometry labs, brought to you by Dr. Peter Lopez.

Definitions

Apheresis

Apheresis collection of the mononuclear cell (MNC) layer has been shown to be a safe and efficient method of collecting the large number of T lymphocytes necessary to initiate CART cell culture. Apheresis involves application of centrifugal force to a continuous or semi continuous flow of anti-coagulated whole blood. As cell layers separate by density, individual layers may be selectively and efficiently removed or replaced. The mononuclear cell layer is located between the dense polymorphonuclear cell / red blood cell layers and the less dense platelet layer. Circulating mature lymphocytes can be found within the MNC layer; therefore, isolation of this layer provides the cells to begin CAR-T cell manufacture. (*1)

Isolation

Selection of specific cell constituents with or without a wash step allows for purification of cell populations prior to seeding CAR-T cell culture. Cell selection is achieved primarily by cell density, cell size or immunophenotypic features. Sedimentation agents, such as Ficoll-Hypaque, use density to purify lymphocytes from MNC products.(*2)


Sources:

(*1) Fesnak, A., Lin, C., Siegel, D. L., & Maus, M. V. (2016). CAR-T Cell Therapies From the Transfusion Medicine Perspective. Transfusion medicine reviews, 30(3), 139–145. https://doi.org/10.1016/j.tmrv.2016.03.001
(*2) Ferrante, A., & Thong, Y. H. (1980). Optimal conditions for simultaneous purification of mononuclear and polymorphonuclear leucocytes from human blood by the Hypaque-Ficoll method. Journal of immunological methods, 36(2), 109–117. https://doi.org/10.1016/0022-1759(80)90036-8