ABSTRACT
The production of cultured red blood cells (cRBC) for transfusion purposes requires large scale cultures and downstream processes to purify enucleated cRBC. The membrane composition, and cholesterol content in particular, are important during proliferation of (pro)erythroblasts and for cRBC quality. Therefore, we tested the requirement for cholesterol in the culture medium during expansion and differentiation of erythroid cultures with respect to proliferation, enucleation and purification by filtration. The low cholesterol level (22 µg/dl) in serum free medium was sufficient to expand (pro)erythroblast cultures. Addition of 2.0 or 5.0 mg/dL of free cholesterol at the start of differentiation induction inhibited enucleation compared to the default condition containing 3.3 mg/dl total cholesterol derived from the addition of Omniplasma to serum free medium. Addition of 5.0 mg/dl cholesterol at day 5 of differentiation did not affect the enucleation process but significantly increased recovery of enucleated cRBC following filtration over leukodepletion filters. The addition of cholesterol at day 5 increased the osmotic resistance of cRBC. In conclusion, cholesterol supplementation after the onset of enucleation improved the robustness of cRBC and increased the yield of enucleated cRBC in the purification process.
Subject(s)
Cholesterol , Culture Media , Erythrocytes , Cholesterol/metabolism , Humans , Erythrocytes/metabolism , Culture Media/chemistry , Cells, Cultured , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Cell Culture Techniques/methods , Erythroblasts/metabolism , Erythroblasts/cytology , Culture Media, Serum-FreeABSTRACT
Non-alcoholic steatohepatitis (NASH) is viewed as the hepatic manifestation of the metabolic syndrome and is a condition hallmarked by lipid accumulation in the liver (steatosis) along with inflammation (hepatitis). Currently, the etiology and mechanisms leading to obesity-induced hepatic inflammation are not clear and, as a consequence, strategies to diagnose or treat NASH in an accurate manner do not exist. In the current review, we put forward the concept of oxidized lipids as a significant risk factor for NASH. We will focus on the contribution of the different types of oxidized lipids as part of the oxidized low-density lipoprotein (oxLDL) to the hepatic inflammatory response. Furthermore, we will elaborate on the underlying mechanisms linking oxLDL to inflammatory responses in the liver and on how these cascades can be used as therapeutic targets to combat NASH. This article is part of a Special Issue entitled: Lipid modification and lipid peroxidation products in innate immunity and inflammation edited by Christoph J. Binder.
Subject(s)
Lipoproteins, LDL/immunology , Lipoproteins, LDL/metabolism , Non-alcoholic Fatty Liver Disease/immunology , Non-alcoholic Fatty Liver Disease/metabolism , Humans , Inflammation/immunology , Inflammation/metabolism , Lipid Metabolism/immunology , Lipid Metabolism/physiology , Liver/immunology , Liver/metabolismABSTRACT
The role of transcranial Doppler ultrasound (TCD) was studied retrospectively in relation to the management of 73 patients with a subarachnoid hemorrhage due to a ruptured intracranial aneurysms. TCD velocity measurements provided very valuable data to assist scheduling of angiography and delayed surgery. Angiography and surgery were planned significantly sooner in the TCD examined group of patients with TCD velocities indicative for the absence of vasospasm than in the group of patients who did not have TCD examinations. The cerebral circulatory resistance index (R), calculated as: (maximum systolic velocity--end diastolic velocity)/maximum systolic velocity, and changes in R did not predict a change in clinical grade or outcome.
