All patients were treated with myeloablative conditioning regimens, including hyper fractionated total body irradiation 1375 cGy over 4 days, followed by thiotepa 5 mg/kg/day i.v. for 2 days and either fludarabine 25 mg/m2/day i.v. for 5 days, or cyclophosphamide 60 mg/kg/day i.v. for 2 days; or busulfan followed by melphalan 70 mg/m2/day i.v. for 2 days, and fludarabine 25 mg/m2/day i.v. for 5 days. TCD of granulocyte colony-stimulating factor-mobilized PBSCs grafts was performed as described previously [8 (link), 9 (link)]. Ex vivo CD34+ selection of hematopoietic progenitor cells was performed using one of two methods as previously described: Isolex 300i Magnetic Cell Separator (Baxter, Deerfield, IL), followed by T cell rosetting with sheep erythrocytes (9 patients), or using the CliniMACS® CD34+ Reagent System (Miltenyi Biotech, Gladbach, Germany). All patients received either equine or rabbit anti-thymocyte globulin (ATG). Patients did not receive any other post-transplant immunosuppressive prophylaxis. All patients received supportive care and prophylaxis against opportunistic infections according to standard guidelines [10 (link)].
Clinimacs cd34 reagent system
Manufactured by Miltenyi Biotec
Sourced in Germany
The CliniMACS CD34 Reagent System is a laboratory equipment for the selection and enrichment of CD34+ cells from human blood or bone marrow samples. It utilizes magnetic beads coated with antibodies targeting the CD34 surface antigen to isolate the desired cell population.
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Lab products found in correlation
17 protocols using clinimacs cd34 reagent system
1
Myeloablative Conditioning and Stem Cell Transplantation
2
CD34+ Cell Isolation and Cryopreservation
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A CliniMACS CD34+ reagent system (Miltenyi Biotec) was used to isolate CD34+ hematopoietic stem/progenitor cells at the Juravinski Cancer Centre (using established SOPs) from the leukapheresis product harvested on the previous day. After magnetic labeling with an anti-human CD34 antibody, the CD34+ hematopoietic cells were enriched using a high-gradient magnetic separation column connected to a closed disposable tubing set. After washing steps, the retained CD34+ hematopoietic cells were eluted by removal of the magnetic field. The degree of purification was monitored by flow cytometry analysis after staining an aliquot of purified product with a two-color fluorescein isothiocyanate (FITC)-CD45/PE-CD34 reagent from the Stem-Kit (Beckman Coulter).
Isolated CD34+ cells were immediately resuspended in 10% Cryoserv DMSO (Bioniche PHARMA) plus 10% autologous plasma and 80% Plasma-Lyte A (Baxter) in CryoStore freezing bags (OriGen Biomedical) and cryopreserved using a controlled-rate freezer (Planer). Following this, the cells were stored in the vapor phase of a liquid nitrogen tank. The cryopreserved CD34+ cells were later transported to our laboratory at UHN or directly to the Philip S. Orsino Cell Processing Facility at Princess Margaret Hospital for LV transduction.
Isolated CD34+ cells were immediately resuspended in 10% Cryoserv DMSO (Bioniche PHARMA) plus 10% autologous plasma and 80% Plasma-Lyte A (Baxter) in CryoStore freezing bags (OriGen Biomedical) and cryopreserved using a controlled-rate freezer (Planer). Following this, the cells were stored in the vapor phase of a liquid nitrogen tank. The cryopreserved CD34+ cells were later transported to our laboratory at UHN or directly to the Philip S. Orsino Cell Processing Facility at Princess Margaret Hospital for LV transduction.
3
G-CSF Mobilized PBSC Graft Depletion
4
Conditioning Regimens for Hematopoietic Cell Transplant
5
Autologous PBSC Mobilization, Selection, and Transplantation
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Peripheral blood stem cells (PBSC) were mobilized by administration of recombinant human G-CSF (from Day 4 until apheresis) after chemotherapy with cyclophosphamide (CYC; 2 × 1–2 g/m² body surface) for 2 consecutive days and collected according to local standards, beginning on day 10. In one patient - due to CYC intolerance -, stem cells were obtained by bone marrow aspiration without previous chemotherapy. We aimed for a cell count > 2 × 106 CD34 + cells/kg bodyweight (BW), and all cells were positively selected via immunomagnetic beads (CliniMACS® CD34 Reagent System, Miltenyi, Bergisch Gladbach, Germany).
Standard pre-transplant conditioning regimen contained of CYC 50 mg/kg bodyweight (day − 5 until day − 2) and T-cell depletion therapy with rabbit antithymocyte globulin (ATG, grafalon®, Neovii Pharmaceuticals AG, Switzerland) in varying doses from 2.5 to 40 mg/kg BW (day − 4 until day − 1), or thiotepa 2 × 5 mg/kg BW (day − 5), ATG 2.5–40 mg/kg BW (day − 4 until day − 1), and CYC 50 mg/kg BW (day − 3 until day − 2) depending on the disease manifestation as described previously [7 (link)]. We used melphalan 100 mg/m² body surface area (BSA) in the one patient intolerant to CYC. Frozen-thawed CD34 + positively selected PBSC were transplanted on day 0. There was no application of G-CSF after reinfusion of autologous stem cells in any patient.
Standard pre-transplant conditioning regimen contained of CYC 50 mg/kg bodyweight (day − 5 until day − 2) and T-cell depletion therapy with rabbit antithymocyte globulin (ATG, grafalon®, Neovii Pharmaceuticals AG, Switzerland) in varying doses from 2.5 to 40 mg/kg BW (day − 4 until day − 1), or thiotepa 2 × 5 mg/kg BW (day − 5), ATG 2.5–40 mg/kg BW (day − 4 until day − 1), and CYC 50 mg/kg BW (day − 3 until day − 2) depending on the disease manifestation as described previously [7 (link)]. We used melphalan 100 mg/m² body surface area (BSA) in the one patient intolerant to CYC. Frozen-thawed CD34 + positively selected PBSC were transplanted on day 0. There was no application of G-CSF after reinfusion of autologous stem cells in any patient.
6
Targeted CD34+ Cell Selection and Viral Monitoring
7
Allo HCT with CD34+ Selection by Age
8
Graft Manipulation and Conditioning Regimens
9
Retrospective Analysis of IC in Allo-HCT
10
Myeloablative Regimen for Multiple Myeloma
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