FACS Core Unit Team

FACS Core Unit Team

Team Leader
Assoc. Prof. Gerhard Fritsch, PhD
Email: gerhard.fritsch(at)ccri.at 

Technicians (For the FACS services)
Ing. Dieter Printz (Facility operator)
Julia Stemberger
Elke Zipperer

The "Laboratory for Clinical Cell Biology and FACS Core Unit” – was established in 1989 and is headed by Assoc. Prof. Gerhard Fritsch. After one year of work, the team managed to isolate blood cells from a mobilised patient for the first microscopical observation. Within the following years, the team optimised the flow cytometric measurements and established routine tools to provide information to clinicians about stem cells as well as all other blood leukocytes. 

Clinical-scale cell manipulation commenced in 1996 and includes cryopreservarion of stem cells as well as cell selection prior to allogeneic stem cell transplantation in case of donor/patient HLA mismatch (see Figure).

Please click here for details about the research activities of the working group “Clinical Cell Biology and FACS Core Unit” 

As a FACS (flow cytometry) core unit, the laboratory presently has at its disposal three flow cytometers for 15 to 19 parameter cell measurements, one of which is a multi-colour device for cell sorting:

•    LSR II: 405, 488 & 635 nm lasers, 15 parameters
•    LSR Fortessa: 405, 488, 561 & 640 nm lasers, 19 parameters
•    FACSAriaFusion: 405, 488, 561 & 640 nm lasers, 19 parameters

All devices are serviced by an experienced technician. In close cooperation with clinicians, St. Anna researchers and other scientists, the FACS Core Unit offers practical and theoretical support for all questions regarding flow cytometry.

Depletion of T-cells from an HLA mismatched donor prior to stem cell transplantation.

Stem cell transplantation (SCT) is the transplantation of haematopoietic stem cells from the donor to a patient. SCT is most often performed for patients with life threatening diseases of the blood and cancer if conventional treatment options failed. For optimal SCT, the tissue (HLA) type of both the donor and the patient should match. Unfortunately, for some patients, an appropriately matched donor is not available. Therefore, mismatched donors are chosen. However, this mismatched situation increases the risk for graft versus host disease (GvHD) in patients. GvHD in patients is induced by potent immune cells (T cells) from the donor that will react against patient´s tissue cells. To overcome that problem, immune cells from the donor have to be depleted from the stem cell product prior to transplantation into the patient. This magnetic depletion strategy is shown in the drawing. 

Services

Diagnostics

Flow cytometric analyses (Together with LabDia)

  • Residual WBC analysis in plasma or platelet concentrates as QC (collaboration with Humanplasma, Vienna)
  • Quantification of WBC in hypoplastic/aplastic blood or bone marrow, or in blood products,
  • Single-platform CD34 analysis in thawed cord blood as QC (since 2010, collaboration with Eurocord Slovakia, Bratislava)
  • Diagnosis of spherocytosis (offered throughout Austria since 2009)
  • Human sperm fertility analysis (since 2010, collaboration with Kinderwunschzentrum Goldenes Kreuz, Vienna)
  • Apoptotic cell quantification based on a recently validated single-platform no-wash 7-colour analysis
  • Quantification of WBC, WBC subtypes as well as CD34 and CD34 subtypes in blood, cord blood, bone marrow, as well as in fresh and thawed PBSC
  • Subtype analysis of T cells (CD4, CD8, alpha-beta, gamma-delta, HLA-DR, CD25, naïve/memory)
  • Analysis of DNA content and cell cycle
  • Multi-color analysis of CFSE-labeled cells to detect cell division  
  • We participate in the recently commenced CD14/18 study, by providing quantitative data regarding the numbers of peripheral CD16- NK cells and CD127- CD4+/CD25+ T cells during the course of radioactive MAB treatment of neuroblastoma patients.
  • Examination of donor and patient blood samples for the presence of virus-specific CD3 positive T cells, using either the cytokine secretion assay (CSA, expression of interferon-gamma by CD4 and CD8 positive T cells upon in-vitro virus-specific antigen stimulation), and/or multimer analysis which is specific for CD3/CD8 positive lymphocytes. 
  • Whereas anti CMV analysis has been routinely used during the last years, new techniques are emerging that address T cell specificity against other clinically important viruses, particularly ADV, EBV and BKV. The type of analysis chosen depends on the availability of respective HLA-specific multimers, but also on the expected percentage of target cells. If necessary, the analysis is preceded by a several day in-vitro stimulation which usually results in a relative enrichment and thus a better detectability of the target cells.

Cellular therapeutic agents for clinical routine

  • Cryopreservation and storage of, and storage logistics for autologous and allogeneic blood products
  • Thawing for reinfusion
  • CD3/19 depletion prior to HLA-mismatch stem cell transplantation
  • Cell manipulations and analyses for DLI
  • Preparation of MNC from blood for extracorporeal photopheresis ("mini ECP") for treatment of GvHD