Isolation of malignant cells from human bone marrow using a discontinuous percoll gradient

Macfarlane, S. D.; Tauro, G. P.; Rendall, K.; Welch, John J.; Ekert, H.

doi:10.1002/ajh.2830220409

Cited by 3 publications

(1 citation statement)

References 7 publications

(7 reference statements)

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“…According to Macfarlane et al [9], the use of discontinuous Percoll gradients offers no advantage over conventional methods in the diagnosis of haematological malignancies. Nevertheless, if suspecting early relapse of AL, this narrow range density gradient method might be better for blast enrichment than other methods, for example, for immunological cell marker studies.…”

Section: Discussionmentioning

confidence: 99%

Human peripheral blood and bone marrow cell separation using density gradient centrifugation on Lymphoprep and Percoll in haematological diseases

Koistinen¹

1987

Scand J Clin Lab Invest

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Koistinen P. Human peripheral blood and bone marrow cell separation using density gradient centrifugation on Lymphoprep and Percoll in haematological diseases. Scand J Clin Lab Invest 1987; 47: 709-714. Density gradient centrifugation on Lymphoprep as described by Boyum [ 11 was used for mononuclear cell separation of peripheral blood (PB) and bone marrow (BM) samples in healthy subjects and in haematological diseases. The cells were analysed morphologically from cytocentrifuge preparations stained with May-Griinwald-Giemsa. At least 200 cells per preparation were counted manually. The separation results of PB samples in healthy subjects were comparable with those described by Boyum. This method was also suitable for the separation of lymphocytes and blast cells in chronic lymphoproliferative diseases and in acute leukaemias, respectively. In other haematological diseases, however, the mononuclear cell fractions of PB samples and especially of BM samples were contaminated with myeloid and erythroid cells. Percoll gradients were used with success for the enrichment of myeloid BM cells and PB reticulocytes. However, absolutely pure myeloid cell fractions from different maturation stages could not be separated. The results indicate that cells to be investigated should be morphologically examined, to avoid erroneous interpretation of biochemical and functional activity of the cells.

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Section: Discussionmentioning

confidence: 99%

Human peripheral blood and bone marrow cell separation using density gradient centrifugation on Lymphoprep and Percoll in haematological diseases

Koistinen¹

1987

Scand J Clin Lab Invest

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Bone marrow transplantation for solid tumors in pediatrics

Moss¹

1995

Cancer Treatment and Research

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Isolation and storage of peripheral blood hematopoietic stem cells for autotransplantation into children with cancer

Takaue

Watanabe

Kawano

et al. 1989

Blood

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Peripheral blood stem cells (PBSC) were collected for autotransplantation by a total of 46 continuous-flow leukaphereses in 17 children with various types of cancer in whom the stem-cell pool had been expanded by chemotherapy. As the cells collected by leukapheresis were contaminated with many visible cell clumps, platelets, and erythrocytes, they were separated from the platelet-rich plasma (PRP) by slow-speed centrifugation and fractionated on a discontinuous gradient of Percoll. All the hematopoietic progenitors (CFU-GM, CFU- GEMM) in the starting samples were recovered at the interface of 40% and 60% Percoll solutions largely free of other cellular components and with a substantial reduction in volume. The separation and freezing procedures could be completed within three hours after obtaining cells by leukapheresis. After their fractionation and storage, these PBSC were shown to be able to reconstitute normal hematopoiesis in ten children with poor prognosis leukemia or neuroblastoma for whom no HLA- compatible marrow donors were available and who had been subjected to marrow-ablative therapy. This separation procedure is simple, efficient, and readily available and can be used for children as a routine procedure for PBSC autotransplantation.

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Isolation of malignant cells from human bone marrow using a discontinuous percoll gradient

Cited by 3 publications

References 7 publications

Human peripheral blood and bone marrow cell separation using density gradient centrifugation on Lymphoprep and Percoll in haematological diseases

Human peripheral blood and bone marrow cell separation using density gradient centrifugation on Lymphoprep and Percoll in haematological diseases

Bone marrow transplantation for solid tumors in pediatrics

Isolation and storage of peripheral blood hematopoietic stem cells for autotransplantation into children with cancer

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