Separation of Mononuclear Bone Marrow Cells using the Cobe 2997 Blood Cell Separator

Faradji, A.; Andreu, G.; Pillier-Loriette, C.; Bohbot, Alain; Nicod, A; Autran, Brigitte; Bergerat, Jean‐Pierre; Rio, Bernard; Leblond, Véronique; Binet, Jacques‐Louis; Zittoun, R; Oberling, F.

doi:10.1111/j.1423-0410.1988.tb05079.x

Cited by 22 publications

(4 citation statements)

References 24 publications

(9 reference statements)

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“…Freezing and thawing cause lysis of RBC and granulocytes. Lysis of granulocytes results in the release of DNA and lysosomal enzymes causing clumping of the thawed BM [6]. To achieve cryopreservation it is necessary to remove RBC, granulocytes and plasma with minimal loss of progenitor cells.…”

Section: Introductionmentioning

confidence: 99%

Automated Processing of Human Bone Marrow Grafts for Transplantation

et al. 1993

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Prior to purging or cryopreservation, we concentrated 21 bone marrow (BM) harvests using a modification of the 'grancollect-protocol' of the Fresenius AS 104 cell separator with the P1-Y set. Within 40-70 min, the initial marrow volume of 1,265 ml (+/- 537 ml) was processed two to three times. A mean of 47% (+/- 21%) of the initial mononuclear cells was recovered in a mean volume of 128 ml (+36 ml). The recovery of clonogenic cells, measured by CFU-GM assays, was 68% (+/- 47%). Red blood cells in the BM concentrates were reduced to 7% (+/- 4%) of the initial number. The procedure was efficient and yielded a BM cell fraction suitable for purging, cryopreservation and transplantation. At this time, 10 of the 21 patients whose BM was processed using this technique have been transplanted. Seven of these 10 patients have been grafted using the BM alone. Three of the 10 patients showed reduced cell viability and colony growth in the thawed BM samples, and therefore obtained BM and peripheral blood-derived stem cells. All transplanted patients showed an evaluable engraftment, achieving 1,000 granulocytes per microliter of peripheral blood in a mean of 18 days.

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Section: Introductionmentioning

confidence: 99%

Automated Processing of Human Bone Marrow Grafts for Transplantation

et al. 1993

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“…The present study with Haemonetics V50 was performed with special reference to very small bone marrow volumes ranging from 200 to 399 ml (with the 100 ml of medium 199 included), which have not been the focus of any previous study. Small marrow volumes have instead been recognized as a serious practical problem [12].…”

Section: Discussionmentioning

confidence: 99%

Bone Marrow Fractionation by the Haemonetics System: Reduction of Red Cell Mass before Marrow Freezing, with Special Reference to Pediatric Marrow Volumes

Saarinen¹,

Lähteenoja²,

Juvonen³

1992

Vox Sanguinis

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For purposes of freezing autologous marrow or transplants of allogeneic marrow with major ABO blood group incompatibility, 54 freshly harvested bone marrows from children of 7-65 kg of weight were depleted of their red cells with the Haemonetics V50 system. The marrow volumes ranged from 230 to 1,145 ml, with 17 small (200-399 ml), 18 intermediate (400-799 ml) and 19 large (800-1,200 ml) volumes. After processing, the median recoveries were: volume 24%, red cell mass 18%, and nucleated cells 75%. In the small marrow volume group, a good nucleated cell recovery was achieved at the expense of red cell depletion. The colony-forming units, granulocytes-macrophages (CFU-GM) were normal after thawing of processed, cryopreserved marrows, and good engraftment of both allogeneic and autologous marrows were achieved. We conclude that marrow processing with the Haemonetics V50 system results in adequate red cell depletion and good nucleated cell recovery without open-air contact of marrow or excessive time consumption. For small marrow volumes, however, the red cell depletion was suboptimal, and a bowl size smaller than 125 ml is desirable for pediatric use.

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“…Physical methods of cell separation include velocity sedimentation, density gradient centrifugation, counterflow centrifugal elutriation, and related techniques (reviewed in Kumar and Lykke, 1984). Velocity sedimentation, using automated or semiautomated cell-separating centrifuges, has been used to separate the red cells and plasma from the nucleated cells in marrow (Gilmore et al, 1983; Faradji et al, 1988) and peripheral blood (Korbling and Martin, 1988;Williams et al, 1990). This technique reduces the volume of material by about 10-fold and recovers 60-80% of nucleated cells.…”

Section: Methods Of Cell Selectionmentioning

confidence: 99%