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Bone Marrow Harvesting and Reinfusion
Published in Adrian P. Gee, BONE MARROW PROCESSING and PURGING, 2020
In the early years of bone marrow harvesting and transplantation, it was deemed necessary to filter aspirated bone marrow and blood, in order to remove fibrin clots and other debris prior to infusion into the recipient, and many centers still adopt this policy. Marrow is commonly sieved through a thin stainless steel mesh, 380-μm pore initially, and subsequently through one of 80 μm size, prior to collection in a sterile vessel containing anticoagulant.2 It is subsequently aspirated into a sterile pack for processing. There is no firm evidence, however, that filtering marrow during aspiration is necessary. Syringes of anticoagulated marrow should be introduced directly into a dry, sterile blood pack into which 5000 units of preservative-free heparin have been introduced. This can be accomplished either by use of a three-way tap attached to the syringe and blood pack by sterile tubing or by injecting the marrow directly into the blood pack by connecting the syringe to a sterile plastic injection port. Contact with the atmosphere is thus kept to a minimum, reducing likelihood of infection. When the bag contains 400 to 500 ml of marrow, it can be sealed, ready either for infusion into the recipient, for purging, or for cryopreservation. If marrow is to be refrigerated for a few hours prior to infusion into the recipient, some centers add 60 ml of citrated phosphate dextrose (CPD), or acid citrate dextrose (ACD), designed to supply nutrients to the stem cells during the time they are out of the body.
Therapeutic apheresis
Published in Jennifer Duguid, Lawrence Tim Goodnough, Michael J. Desmond, Transfusion Medicine in Practice, 2020
The use of citrate anticoagulation can lead to mild hypocalcemia characterized by tingling, oral paresthesias, or chest discomfort. Citrate reactions are usually mild and easily managed by slowing the infusion rate, increasing the acid-citrate-dextrose (ACD) ratio, or administering oral calcium. More severe reactions may be treated or prevented with intravenous calcium. Calcium chloride may be added to the replacement fluid (200 mg/l of 5% albumin or 3–6% HES) or given as a slow intravenous infusion (200 mg, diluted to at least 20 mg/ml and infused over 2 minutes) when replacing with FFP. Particular care must be taken with patients with liver failure and impaired ability to metabolize citrate.
A Survey of Newer Gene Probing Techniques
Published in Victor A. Bernstam, Pocket Guide to GENE LEVEL DIAGNOSTICS in Clinical Practice, 2019
In the isolation of nucleic acids from blood samples, the acid citrate dextrose (ACD) anticoagulant appears to be superior to ethylenediamine-tetraacetic acid (EDTA) and heparin in not affecting the yield of DNA, even from frozen blood kept in ACD for 5 days at 23°Callowing unaffected restriction patterns to be obtained with three enzymes (EcoRI, HindIII, and XbaI)
Efficacy of autologous platelet-rich plasma therapy versus topical Minoxidil in men with moderate androgenetic alopecia: a randomized open-label trial
Published in Journal of Dermatological Treatment, 2023
Mithinkumar Balasundaram, Rashmi Kumari, Sivaranjini Ramassamy
Our institution’s PRP preparation protocol is standardized and has consistently resulted in a platelet yield of 3–5 times the baseline value. The platelet concentration in the PRP ranged from 9 to 10.2 lakhs/µL. The yield is a pure PRP, i.e. the percentage of platelets in the PRP compared with RBC and leucocytes ranges from 70 to 90%. We collected whole blood (19 ml) and transferred it to 2 Acid citrate dextrose-containing vacutainer tubes, 8.5 ml each. The sample was centrifuged at 400 g for 10 min at room temperature. At the end of the first spin, we transferred the upper and intermediate layers to plain 8.5 ml tubes, without disturbing the bottom layer of RBCs. We performed the second centrifugation at 900 g for 10 min to concentrate the platelets and WBCs in the PRP. At the end of the second spin, we discarded two-thirds of the supernatant and gently suspended the lower third (PRP). At the end, we obtained almost 2 ml of PRP for injection.
High-efficiency unassisted transfection of platelets with naked double-stranded miRNAs modulates signal-activated translation and platelet function
Published in Platelets, 2021
Sophia Lazar, Jeremy G.T. Wurtzel, Xi Chen, Peisong Ma, Lawrence E. Goldfinger
Human blood was obtained by venipuncture from a pool of healthy volunteers in a one-sixth volume of acid-citrate-dextrose. Red blood cells were removed by centrifugation at 230xg for 20 min at room temperature. Platelet-rich plasma was recovered, and platelets were pelleted at 800xg for 10 min at room temperature. The platelet pellet was suspended in Tyrode’s buffer (138 mM NaCl, 2.7 mM KCl, 1 mM MgCl2, 3 mM NaH2PO4, 5 mM glucose, 10 mM Hepes, pH 7.4 in distilled, DNase/RNase-free water with 0.01 units/ml of apyrase added fresh) containing 1 mg/ml bovine serum albumin (BSA). Leukocyte depletion was achieved by incubation of platelet suspensions with α-CD45- and α-CD235a-conjugated magnetic microbeads at 10 μl each per 10 ml blood, for 15 min with mixing at 4ºC, followed by bead extraction with MACS separation LD columns according to the manufacturer’s instructions. Leukocyte depletion was confirmed by RNA extraction from a sample of the pre- and post-depleted platelet supernatants, followed by cDNA synthesis and PCR for CD45 mRNA as described below; data were obtained only from platelet samples with no detectable CD45 expression. Approval for this study was obtained from the Institutional Review Board of Thomas Jefferson University. Written informed consent was obtained after the nature and possible consequences of the study were explained.
Comparison of the GPVI inhibitors losartan and honokiol
Published in Platelets, 2020
Marie-Blanche Onselaer, Magdolna Nagy, Chiara Pallini, Jeremy A Pike, Gina Perrella, Lourdes Garcia Quintanilla, Johannes A Eble, Natalie S. Poulter, Johan W.M. Heemskerk, Steve P Watson
Venous blood was taken from healthy volunteer using 3.8% (v/v) sodium citrate (1:9) as the anti-coagulant with informed consent according to the guidelines of the local ethics committee (ERN_11-0175). All steps of this study complied with the ethical principles according to the Declaration of Helsinki. Acid Citrate Dextrose (ACD, 1:10) was added to the blood. Platelet-rich plasma (PRP) was obtained by centrifugation at 200 g for 20 min at room temperature. Washed platelets were obtained by centrifugation at 1000 g for 10 min at room temperature using prostacyclin (2.8 μM) and resuspended in modified Tyrode’s-HEPES buffer (134 mMNaCl, 0.34 mM Na2HPO4, 2.9 mMKCl, 12 mM NaHCO3, 20 mM HEPES, 5 mM glucose, 1 mM MgCl2; pH7.3) Washed platelets were used at 2 × 107/ml for static adhesion or 5 × 108/ml for other studies.