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Cytotherapy. 1999;1(6):469-77.

T-cell depletion of allogeneic bone marrow grafts: enrichment of mononuclear cells using the COBE Spectra cell processor, followed by immunoselection of CD34(+) cells.

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1
Centre de Thérapie Cellulaire, Institut Paoli-Calmettes, Centre Régional de Lutte Contre le Cancer Provence-Alpes-Côte d'Azur, 232 Boulevard de Ste Marguerite, BP 156. 13273 Marseille Cedex 9, France.

Abstract

BACKGROUND:

Enrichment in mononuclear cells (MNC) is a prerequisite for CD34(+) cell selection from BM allografts. However, centrifugation over a density gradient (Ficoll), does not comply with good manufacturing practice (GMP), because Ficoll is not a clinical grade reagent. We evaluated the COBE Spectra cell separator as an alternative and evaluated the recovery of MNC and CD34(+) cells. CD34(+) cell selection was then performed using either the Nexell Isolex 300i or the CEPRATE SC (CellPro) device.

METHODS:

20 allogeneic BM grafts were processed, with an initial enrichment of MNC using the COBE Spectra processor, followed by immunoselection of CD34(+) cells (13 Isolex 300i and seven CEPRATE SC). We evaluated total nucleated cells (TNC), MNC and CD34(+) cells recoveries when using COBE Spectra and compared CD34(+) cells recovery and T-cell depletion when using Isolex 300i or CEPRATE SC.

RESULTS:

Median recoveries of the product using the COBE Spectra were 31.2% (18.1-68.9%) for TNC, 76.9% (29.5-148.1%) for MNC and 79.6% (60.9-191%) for CD34(+) cells. Median recoveries of CD34(+) cells were 39.4% (15.1-75.8%) and 36.7% (26.2-64.3%) with the Isolex 300i and CEPRATE SC devices, respectively. CD34(+) purity was 85% (64.4-94.5%) and 84.4% (66.8-86.8%). Colony-forming unit granulocyte/macrophage (CFU-GM) recoveries were 10.9% (0.2-152%) and 36% (0.0-67.3%). Median log depletion for T-cell numbers, B-cell numbers and natural killer (NK) cell numbers were 2.8 log (2.1-3.8), 2.0log (0.95-3.1) and 3.1log (2.40-4.8), respectively.

DISCUSSION:

Processing with COBE Spectra produces a cell population that is suitable for CD34(+) cell selection. The Isolex 300i and CEPRATE SC devices demonstrate similar performances. The two-step procedure allows allo-BMT in patients with high risk of GvHD.

PMID:
20443236
[Indexed for MEDLINE]
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