A European reference protocol for quality assessment and clinical validation of autologous haematopoietic blood progenitor and stem cell grafts

Abstract

Summary:Recently, the regulatory authorities have begun to show interest in haematopoietic stem cell products. On a professional rather than a regulatory basis, the International Society for Hematotherapy and Graft Engineering (ISHAGE) has established the Foundation for the Accreditation of Haematopoietic Cell Therapy (FACHT), which has drawn up guidelines for standards and accreditation of such activity. In Europe, the regulatory environment with regard to haematopoietic stem cell grafts, processing and storage are currently less stringent. However, in 1998 the European Joint Accreditation Committee Euro-ISHAGE/EBMT (JACIE) prepared a regulatory document 'Standards for Blood and Marrow Progenitor Cell Collection, Processing and Transplantation' which was approved by the EBMT General Assembly. The major objectives were to promote quality of medical and laboratory practice in haematopoietic progenitor cell transplantation. The standards extend and detail the pre-existing activity of EBMT centres including all phases of collection, processing and administration of these cells. This is the platform for the proposed reference protocol for CD34 ϩ cell enumeration and clinical validation of quality assessment to ensure that appropriate standards of work and product quality are established and will be maintained. Bone Marrow Transplantation (2001) 27, 463-470.

HLA-identical stem cell transplantation: is there an optimal CD34 cell dose?

“…There are a variety of different techniques that are employed among clinical laboratories, including the ISHAGE, Milan, and Norway protocols, along with commercial reagents such as ProCOUNTt from Becton Dickinson and Stem Kitt from Beckman Coulter, for quantitative CD34 assessment. [38][39][40][41][42][43][44] Each of these methods utilizes slightly different staining combinations and/or gating strategies that can result in somewhat different final values. It is important to keep in mind that when a specific cell dose is referred to in the published literature, it may not correspond exactly to the same dose that would be determined by another clinical site using a different procedure.…”

Number of viable CD34+ cells reinfused predicts engraftment in autologous hematopoietic stem cell transplantation

“…13 The dual platform method has been well validated in several clinical studies that included rates of hematopoietic reconstitution. 12,16,17 Guidelines for CD34 + cell enumeration by flow cytometry have been prepared by ISHAGE, 15 the European Bone Marrow Transplant Group 18 and the British Committee for Standards in Haematology 19 supporting flow cytometry as the reference standard for CD34 + cell enumeration. The simpler single platform method was evaluated in 72 patients at a single institution and displayed excellent correlation with the dual platform method.…”

Priming with r-metHuSCF and filgrastim or chemotherapy and filgrastim in patients with malignant lymphomas: a randomized phase II pilot study of mobilization and engraftment

“…8,[27][28][29] The frozen samples were thawed in a 37 1C water bath and washed once in a PBS solution containing DNAase (100 mL Buffer A, 0.5 mL MgCl 2 500 mM þ 2.5 mL DNAase (Pulmozyme 1 mg/mL)) to prevent clumping and cell loss caused by sticky DNA fragments from dead cells, and then washed once in buffer A (500 mL PBS, 2.5 g BSA þ 10 mL Na-EDTA stock solution pH 5.5-5.7) and adjusted to a cell concentration of 2 Â 10 7 cells per mL. Samples of 50 mL were mixed with 50 mL Ab solutions and incubated for 30 min at 4 1C, then washed twice in standard buffer, resuspended to a volume of 3-400 mL and analyzed immediately.…”

“…[3][4] One variable of major effect for post transplant care is the number of PBPCs harvested. [5][6][7][8] Therefore, several clinical studies have aimed to identify priming regimens that improve progenitor and stem cell mobilizations and collections without increased toxicity. Frequently, filgrastim (recombinant methionyl human G-CSF (r-metHuG-CSF)) is administrated alone; however, filgrastim combined with chemotherapy has proven more effective in the context of CD34 þ cell numbers harvested, [9][10][11] and this combination is considered the gold standard for priming and stem cell mobilization in relapsed malignant lymphoma.…”