Report on the European Workshop on Peripheral Blood Stem Cell Determination and Standardization—Mulhouse, France, February 6-8 and 14-15, 1992

Wunder, E.; Sovalat, H.; Fritsch, Gerhard; Silvestri, Federico; Hénon, Philippe; Serke, Stefan

doi:10.1089/scd.1.1992.1.131

Cited by 65 publications

(48 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Flow cytometry represents a valuable tool to quantitate the progenitors in mobilized peripheral blood and to evaluate in real time the engraftment potential of the cytapheresis product. 11 In this regard, several assays for CD34 + cell enumeration have been proposed, [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25] but lack of homogeneous procedures has resulted in the generation of controversial data. Historically, the first attempt at enumerating CD34 + progenitors by flow cytometry was based on the evaluation of HPCA-2 FITC/PE positive cells with a low light scatter (SSC), after initial forward vs SSC live gating excluding debris, platelets, erythrocytes and cell aggregates.…”

mentioning

confidence: 99%

“…12,13 This very simple protocol was subsequently validated in three Nordic workshops involving 28 laboratories. 17 The more recently proposed ISHAGE protocol 16 is based upon the addition of a second antibody staining with CD45 FITC to exclude undesired populations and a sequential gating strategy to select CD34 + CD45 +dim cells which are supposed to be the genuine hematopoietic precursors. We developed in our laboratory a simplified method for CD34 + cell enumeration 21 and compared it with the Milan/Mulhouse 14 and ISHAGE protocols.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Enumeration of CD34+ hematopoietic progenitor cells for clinical transplantation: comparison of three different methods

Venditti

Battaglia

Poeta

et al. 1999

Bone Marrow Transplant

View full text Add to dashboard Cite

Summary:Three different methods for determination of CD34 ؉ cells in G-CSF-mobilized peripheral blood were compared. The methods were: the Milan/Mulhouse protocol, the ISHAGE guidelines for CD34 ؉ cells enumeration and our own protocol. The procedure we have adopted is essentially a Milan/Mulhouse protocolderived methodology combined with a multiparametric approach using the PAINT-A-GATE software analysis program. The samples were collected from 70 patients affected by acute leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, myeloma and breast cancer who were scheduled to receive autologous PBSC transplantation. PBSC collection was performed following mobilization with subcutaneous G-CSF at 5-10 g/kg/day. A minimum target of 2 × 10 6 /kg CD34 ؉ cells was considered an acceptable harvest to ensure a safe transplant. On average, three aphereses per patient were performed and a total of 204 apheresis samples were analyzed. Regression analysis of the percentage and absolute number of CD34 ؉ cells, as calculated with each method, achieved an excellent correlation in spite of methodological differences. In fact, both CD34 ؉dim and CD34 ؉ CD45 ؊ events were included in our gating strategy. In the setting of a triple staining associating CD34, CD38 and CD45, we identified a variable fraction of CD34 ؉ CD38 ؉ CD45 ؊ cells which would be otherwise undetected due to its CD45 negativity. To this end, we used a new technology referred to as laser-scanning cytometry (LSC) which allowed the isolation and morphological identification of CD34 ؉ CD45 ؊ cells. By comparing CD34 ؉ CD45 ؉ and CD34 ؉ CD45 ؊ cells, we found that they share a common morphology, thus confirming the hypothesis that the latter are to be considered for CD34 ؉ cell calculation. The median number of CD34 ؉ cells/kg, as calculated by the three methods, was: 4.79 × 10 6 /kg (range 1-570) for the Milan/Mulhouse protocol, 3.9 ؋ 10 6 /kg (range 0.8-498) for the ISHAGE one, and 5.17 ؋ 10 6 /kg (range 2-599) for our protocol. The median time to ANC and PLT engraftment was 11Correspondence: Dr A Venditti, Cattedra di Ematologia, Università 'Tor Vergata', Divisione di Ematologia, Ospedale S Eugenio, Piazzale Umanesimo 10, 00144 Rome, Italy Received 1 February 1999; accepted 29 June 1999 (range 9-24) and 20 (range 10-70) days, respectively. Our protocol achieved the best correlation between CD34 ؉ cells/kg and time to ANC/PLT recovery according to the Spearman's rank test (r ‫؍‬ ؊40 and P Ͻ 0.015 for ANC, r ‫؍‬ ؊46 and P ‫؍‬ 0.005 for PLT). We conclude that (1) CD45 does not appear the ideal partner of HPCA-2 for determination of hematopoietic progenitors in mobilized peripheral blood; and (2) for clinical application, a single staining with 8G12 appears simple, reliable and feasible when rigorous procedures for sample preparation and acquisition are followed and an adequate software for multiparametric analysis is available.

show abstract

mentioning

confidence: 99%

mentioning

confidence: 99%

Enumeration of CD34+ hematopoietic progenitor cells for clinical transplantation: comparison of three different methods

Venditti

Battaglia

Poeta

et al. 1999

Bone Marrow Transplant

View full text Add to dashboard Cite

show abstract

“…The resulting Milan/Mulhouse manual is still a valuable procedure reference in laboratory practice. 9,10 The subsequent years have resulted in several publications about the standardisation of flow cytometry analysis [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25] as well as reports on the clinical implication of the enumeration of CD34 ϩ cells. 24,[26][27][28][29][30][31][32][33][34][35][36][37] The CD34 antigen is stage-specific and identifies cells in the early stages of haemopoietic differentiation.…”

Section: First Step In Quality Assessment Was Cd34 Standardisationmentioning

confidence: 99%

“…In the first phase, the CD34 technique was established in the laboratory and analysed for specificity, sensitivity, reproducibility and accuracy. [5][6][7][8][9][10][16][17][18][19][20][21][22][23][24][25][26] The subsequent second phase, documented a likely clinical influence by single centres analysing retrospective material/data. [27][28][29][30][31][32][33][34]36,37 The third phase prepared convincing single centre prospective evaluation [51][52][53][54][55][56][57][58] evolving into the most important phase four, a multicentre prospective evaluation based upon important clinical end-points (Table 2).…”

Section: The Final Step In Quality Assessment Is Clinical Validationmentioning

confidence: 99%

“…13,15,17,63 The number of CD34 ϩ events present in the low SSC region is still calculated on the basis of a minimum population in the range of 50 to 100 events positively stained by a CD34 class III antibody. 5,6,10,15,64 The percentage of CD34 ϩ cells can then be calculated by the number of CD34 ϩ events divided by the denominator (ϭCD45 ϩ leukocytes). The total number of CD34 ϩ cells per volume can subsequently be calculated by multiplication of the CD34% and the leukocyte count obtained from a haematology analyzer (the dual platform technique).…”

Section: The Final Step In Quality Assessment Is Clinical Validationmentioning

confidence: 99%

See 1 more Smart Citation

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

Serke

Johnsen

2001

Bone Marrow Transplant

View full text Add to dashboard Cite

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.

show abstract

Mobilization, collection, and characterization of peripheral blood hemopoietic progenitors after chemotherapy with epirubicin, paclitaxel, and granulocyte-colony stimulating factor administered to patients with metastatic breast carcinoma

Bengala

Pazzagli

Tibaldi

et al. 1999

Cancer

View full text Add to dashboard Cite

Report on the European Workshop on Peripheral Blood Stem Cell Determination and Standardization—Mulhouse, France, February 6-8 and 14-15, 1992

Cited by 65 publications

References 8 publications

Enumeration of CD34+ hematopoietic progenitor cells for clinical transplantation: comparison of three different methods

Enumeration of CD34+ hematopoietic progenitor cells for clinical transplantation: comparison of three different methods

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

Mobilization, collection, and characterization of peripheral blood hemopoietic progenitors after chemotherapy with epirubicin, paclitaxel, and granulocyte-colony stimulating factor administered to patients with metastatic breast carcinoma

Contact Info

Product

Resources

About