Red blood units collected from bone marrow harvests after mononuclear cell selection qualify for autologous use

Abstract

Background After large volume bone marrow (BM) harvest, donors and patients can develop severe anaemia, because collected BM can contain up to 20% of their red cell mass. In a prospective analysis, we investigated the feasibility to recover red blood cells (RBCs) from the harvested BM and investigated whether these RBC units meet the quality requirements of the European Council.Patients and Methods From 19 patients (median age 51 yrs, range 31-77) with acute myocardial infarction, who participated in the MYSTAR study, a median volume of 1299 ml (range, 700-1870 ml) BM was collected. During BM processing, mononuclear cells (MNC) were separated using the Cobe SpectraÔ apheresis system and the residual RBCs were collected in a separate bag. The quality of the collected RBCs was assessed by measuring LDH, free haemoglobin, potassium and lactate. Haemolysis was calculated and the intracellular concentration of ATP, ADP, AMP was determined by HPLC.Results RBC units recovered from BM after MNC separation had a mean volume of 312 ± 95 ml with a haematocrit of 47 ± 8AE9%, a haemoglobin content of 51 ± 15 g per unit, a haemolysis of 0AE15 ± 0AE005%, a pH of 6AE8 ± 0AE007 and an intracellular ATP concentration of 135 pmol ⁄ 10 6 RBC ± 41, which is comparable with freshly collected packed red blood cells (PRBCs).Conclusion RBCs, collected from bone marrow harvests, can be used for autologous blood support to minimize allogeneic blood transfusions in donors and patients after large volume BM donation.Key words: blood sparing measurements, bone marrow harvest, bone marrow processing, salvage of red blood cells. IntroductionLarge volume bone marrow (BM) harvest is an alternative to G-CSF-induced peripheral blood stem cell collection in patients or donors who are poor mobilizers, in the paediatric setting or in the regenerative medicine, when patients with myocardial infarction are intended to receive stem cells intracardially [1][2][3][4]. Donors and patients are at risk to develop a severe anaemia because collected BM can contain up to 20% of their red cell mass. To reduce the risk of anaemia after BM donation, at least one unit of autologous blood should be collected prior to the harvest procedure [5]. Alternatively, certain clinicians place the donors on supplemental iron plus rhEPO, transfuse their donors with allogeneic RBC units or tolerate anaemia when not severe [6]. Previous studies have shown that a recovery of approximately 90% of the harvested BM-RBCs decreased the homologous transfusion requirement of healthy BM donors significantly [7]. When paediatric BM donors were investigated and BM-RBCs were separated by density gradient centrifugation using Ficoll-Hypaque, similar results were In a prospective analysis, we assessed the extent of cell lesions occurring during manipulation by measuring the release of lactate dehydrogenase (LDH), free haemoglobin (fHb), potassium (K+) and lactate at each manipulation step. Special attention was paid to intracellular purine content by measuring ATP, ADP, AMP, as a surrogate mar...

Prolonged post‐thaw shelf life of red cells frozen without prefreeze removal of excess glycerol

“…Potassium leakage and haemolysis in the modified method are reduced, indicating that the modified freeze, thaw, wash method results in less red cell damage than the original HGM method as proposed by Valeri. Although the exact reason for this difference remains to be elucidated, most probably, skipping the centrifugation step in the glycerolization procedure plays an important role in minimizing shear stress on the red cells , resulting in less damage, and thus better storable red cells. A number of studies have looked at the effect that haematocrit may have on the freezing response of RBCs .…”

“…This centrifugation step is not only laborious, but also induces a risk of breakage of the bag. In addition, centrifugation force might have a negative effect on the integrity of red cells .…”

Cryopreservation of red blood cell units with a modified method of glycerolization and deglycerolization with the ACP 215 device complies with American and European requirements

“…The impact of shear stress caused by centrifugation is well known. 1,2,6,15 A major drawback of our study is the lack of a conventionally treated control group, but this study was conducted to provide evidence that the modified method is an alternative for all centers lacking the appropriate equipment, as we do. However, the Technical Manual of the AABB and the European guidelines provide clear requirements for frozen and thawed RBC units.…”

“…The influence of RBC distress on the quality of RBCs during the collection process and the extent of storage lesions during liquid storage (4°C ± 2°C) is well known. 1,2 It has been shown previously that the length of centrifugation and especially the gravitational force play an important role in the impairment of RBCs. 1 High gravitational forces or a long exposure time results in accelerated intracellular ATP degradation and consumption of glucose during storage as a result of increased membrane repair processes.…”

“…1,2 It has been shown previously that the length of centrifugation and especially the gravitational force play an important role in the impairment of RBCs. 1 High gravitational forces or a long exposure time results in accelerated intracellular ATP degradation and consumption of glucose during storage as a result of increased membrane repair processes. Subzero storage of RBC units reduces erythrocyte metabolism and allows a frozen storage period of at least 10 years.…”