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Fully revised for the fifth edition, this outstanding reference on bone marrow transplantation is an essential, field-leading resource. Du kanske gillar. Lifespan David Sinclair Inbunden. Ladda ned. Spara som favorit. Laddas ned direkt. Each column consisted of g of silica contained in a plastic cartridge with a void volume of mL. At any rate, as highlighted by the authors, the most interesting thing from the laboratory point of view was that immunoadsorption was less effective than plasma exchange at removing antibodies.

The authors assumed that the reason for the lesser effectiveness was probably related to the nature of the synthetic antigen and physical conformation of the immunoadsorbent material A method certainly less well known than plasma exchange and classic immunoadsorption in vivo , but which should be mentioned, is cryofiltration. This method, which has also been used in major ABO-incompatible bone marrow transplants, was well described by Akio Kawamura et al In this system, the key elements are two filters and the use of heparin.

The pore size of this filter is 0. The separated plasma is then led to the second filter with a pore size of 0. Heparinisation plays a very important part in the production of cryogel.

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First, heparin becomes a core of EDA-positive fibronectin, plasma fibronectin, and a complex of fibrinogen as an adsorbent. Various kinds of high molecular weight proteins, immune complexes, globulins, and medium molecular weight proteins such as amyloid proteins also loosely combine with the outer surfaces of these complexes.

Although this was an interesting and elegant method, from a technical point of view, for removing antibodies, unfortunately, its use in this work was described for only one patient in the case of bone marrow transplantation with major ABO incompatibility and for this reason it is difficult to evaluate its efficacy in the clinical field. Certainly in the year in which the work was published , medical insurance in Japan only covered treatment for ABO-mismatched kidney transplants and not bone marrow transplants, making these latter transplants prohibitively expensive it should be noted that the authors of this review do not know the current Japanese legislation.

This was the reason why Kawamura et al. In this method, the blood is drawn from an antecubital vein, citrated, and plasma is separated using a rotating membrane. Rabitsch et al. This method, unlike the previous ones described, is therefore applied in the post-transplant context. As the number of patients treated with this method was small, the authors suggested caution with offering this tool in routine clinical practice for the treatment of patients with persisting isohaemagglutinins due to major ABO incompatibility after bone marrow transplantation 31 — A brief consideration: difficulties with venous access, anticoagulant toxicity, vascular volume changes, mild platelet depletion, and the risk of infection must be addressed with methods such as plasma exchange or immunoadsorption.

Such risks may be untoward, particularly in severely neutropenic and thrombocytopenic patients with aplastic anaemia. Moreover, the risk of haemolysis remains common to all antibody-depletion techniques used for ABO-incompatible bone marrow transplants. In addition, ABO-incompatible erythrocytes must still be infused into patients with diminished, but still detectable, haemagglutinins with the risk of acute haemolysis.

Reich LM et al. Specifically, in six bone marrow transplants, the RBC depletion was accomplished by the addition of hydroxyethyl starch to the marrow at a ratio of Unfortunately, there was no information on the percentage of RBC depletion Continuing our overview, in a paper published in , Braine HG et al. Before processing with the Haemonetics Model cell separator, the bone marrow was collected, anticoagulated with heparin and diluted with TC-l99 a tissue culture medium and then the final marrow-tissue culture mixture was filtered through stainless steel screens.

In order to improve the efficiency of depletion of erythrocytes, Jones HM et al. Jin NR et al. The median residual marrow RBC volume was 1. Falkenburg JHF et al. The procedure was performed in glass bottles and it was important to evaluate the haematocrit exactly in order to prevent aspecific cell loss during centrifugation of the suspension.

This technique was time consuming 4—5 hours and could be associated with infection e.

Dallas Hope: Bone Marrow Transplant Process Explained — Be The Match

A different method to remove RBC from donor bone marrow, based on sedimentation, was used by a group led by Zintl F. In their study, donor erythrocytes were eliminated from bone marrow in seven ABO-incompatible bone marrow transplants by gravity sedimentation mediated by dextran amidotrizoate. With this technique, the whole bone marrow processing was carried out in a closed system manner Unfortunately, in our opinion the details of the system were poorly described, precluding a full evaluation of its potential. Before being processing with a CS Blood Cell Separator, the bone marrow from three donors, for major ABO-incompatible transplantation, was centrifuged to remove fat.

It is important to highlight that this procedure was not time-consuming, being completed within 60—90 minutes. These data were related to the processing of bone marrow in association with cryopreservation and successful autologous bone marrow transplantation and also in association with all allogeneic bone marrow transplants including minor ABO-incompatibility , because there were no results associated only with allogeneic bone marrow transplantation in the presence of major ABO-incompatibility.

This pioneering study was one of the first opening the way to the use of automated, closed systems for the processing of bone marrow, in a clinical setting, in order to remove RBC for allogeneic bone marrow transplantation in the case of major ABO-incompatibility. In fact, this paper describes a technique for RBC removal in major ABO-incompatible bone marrow transplantation requiring two centrifugation steps, special blood bags and a mechanical device to separate the buffy coat from the RBC within the bag.

Thus, no cell separator was needed and no separation solutions had to be added. However, as mentioned by the authors, experience was necessary to judge where to separate the RBC from the buffy coat since, depending on the haematocrit, the buffy coat layer was raised or lowered during the closing of the clamps of the special bags To avoid problems related to operators and to standardise the methods of bone marrow processing, since blood cell separators have been widely used in clinical routine. In fact, concentration of bone marrow mononuclear cells with a cell separator was first described in using the Haemonetics system These devices enable recovery of increased numbers of progenitor cells with selective elimination of RBC and granulocytes.

Moreover, recovery of progenitor cells can be further improved by the addition of density separation media such as Ficoll during processing The only published data on this machine concern early experiences on autologous bone marrow processing and a few cases of manipulations for ABO-mismatch. In a study published in by Zingsem J et al. In this study, conducted between January and June , 23 bone marrow harvests were processed. For 21 patients, the authors described autologous bone marrow processing. The remaining two cases were ABO-incompatible allogeneic donors.

The mean RBC reduction was As suggested by Schwella N et al. In another study, Schwella et al. In a second study, the same team processed 43 autografts using the Fresenius AS cell separator and recovered medians of This medical device is a continuous flow cell separator that isolates cells initially through centrifugation and the formation of a buffy layer. In fact, this investigation was performed to substitute the Fenwal CS system by the Amicus system because the former was taken off the market by the manufacturer in Austria.

Focusing on the Amicus system, Witt V et al. No major modifications were made to the Amicus system. The cycle volume was set to approximately the volume of the harvested bone marrow.


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The number of cycles was set to process approximately four times the volume of the harvested bone marrow, in a median four cycles minimum 3, maximum 8. The authors reported that the Amicus system performed well in RBC depletion of bone marrow for allogeneic transplantation as compared to the Fenwal CS system and concluded that the Amicus system could be employed for the automated RBC and volume depletion of bone marrow 17 , In the overview of automated systems for the processing of the bone marrow, we feel duty-bound to cite the Sepax system Biosafe SA, Eysins, Switzerland.

Sepax was the first automated technology to be specifically developed for use in stem cell banks, allowing processing of various cellular products with a wide range of input product volumes.

Manual of Stem Cell and Bone Marrow Transplantation / Edition 2

The use of the Sepax system does, however, extend beyond stem cell banking. In fact, Sepax cell processing technology has been adopted in leading clinical research studies and is currently widely used for processing various cellular products, including bone marrow. Specifically, in the context of transfusion medicine, the Sepax S the second version is currently present on the market is frequently used for volume reduction of haematopoietic progenitor cells from bone marrow HPC-M and from apheresis HPC-A , and for the removal of dimethylsulphoxide from HPC-A units before transplantation.

The Sepax S cellular-processing system is a cellular centrifugation instrument that automatically processes blood or blood components in a closed, sterile environment. This system uses rotating syringe technology that provides both separation through rotation of the centrifugation chamber and component transfer through displacement of the syringe piston, using the dedicated CS In , the group led by G.

The volume reduction protocol was designed to remove RBC and plasma, while maintaining a good recovery of mononuclear cells. Twenty-seven procedures were carried out on concentrates collected from 21 patients undergoing autologous transplantation and from six donors for allogeneic transplantation, in the presence of major ABO-incompatibility. Focusing on the processing of bone marrow from donors for allogeneic transplants, the automated procedure resulted in a satisfactory median recovery of It is important to highlight that one advantage of the Sepax S automated method in comparison with the classical manual approaches is the possibility of carrying out the entire processing cycle in a contamination-controlled, closed environment, in accordance with Good Manufacturing Practice guidelines.

Major ABO incompatibility is not a barrier to successful bone marrow transplantation. However, there are potential risks, such as acute or delayed haemolytic reactions, delayed recovery of haematopoiesis and graft rejection.

Manual of Stem Cell and Bone Marrow Transplantation by Joseph H. Antin

In order to avoid these problems, anti-red blood cell antibodies in recipients or red blood cells from the donor marrow are generally removed prior to the bone marrow infusion Summarising donor bone marrow processing, we can say that early attempts to process bone marrow used manual techniques. Although effective, manual methods are, by nature of the excessive handling and exposure to the environment, more likely to result in bacterial contamination of the bone marrow. Moreover, inclusion of unwanted components, such as neutrophils and platelets, is a problem when using manual buffy coat preparation.

In addition, manual techniques require considerable technical expertise and these techniques are inefficient for large volumes of bone marrow 65 — It is important to note that techniques using density gradient reagents, such as Ficoll-Hypaque, can be associated with potential cell injury from these reagents based on the demonstration of a significant reduction in tritiated thymidine incorporation and loss of lymphocyte viability following culture with Ficoll-Hypaque. Moreover, reports from several transplant centres have described longer periods of aplasia and even graft failure after transplantation of bone marrow separated on Ficoll gradients 11 , Physical methods of cell separation include velocity sedimentation, centrifugation using semi-automated and automated devices with or without density gradient materials, counterflow centrifugal elutriation, and related techniques.

The Authors declare no conflicts of interest. National Center for Biotechnology Information , U. Journal List Blood Transfus v. Blood Transfus.

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Author information Article notes Copyright and License information Disclaimer. Received Apr 29; Accepted Aug 7. Keywords: haematopoietic stem cells, bone marrow, ABO-incompatibility. This article has been cited by other articles in PMC. What are the historical approaches to avoid haemolytic reactions and related problems in major ABO-incompatible bone marrow transplantation? Focus on the recipient: the removal of anti-red blood cell antibodies In , Gale RP and colleagues reported one of the first investigations of the effectiveness of plasma exchange and antibody absorption in vivo in preparing recipients for ABO-incompatible bone marrow transplants.

Focus on the donor: manipulation of the bone marrow prior to infusion into the recipient A brief consideration: difficulties with venous access, anticoagulant toxicity, vascular volume changes, mild platelet depletion, and the risk of infection must be addressed with methods such as plasma exchange or immunoadsorption. Zingsem J, et al. Open in a separate window.

Concluding remarks Major ABO incompatibility is not a barrier to successful bone marrow transplantation. Footnotes The Authors declare no conflicts of interest. References 1. Dodds A. ABO incompatibility and blood product support. In: Atkinson K, editor.