Carotenoid and tocopherol concentrations in plasma, peripheral blood mononuclear cells, and red blood cells after long-term beta-carotene supplementation in men

Abstract The regulation of erythroid burst-colony formation was studied in cultures of human peripheral blood mononuclear cells. Numbers of erythropoietin-stimulated colonies obtainable from the cells in response to various treatments were compared. One-day preincubation of the cells with phytohemagglutinin (PHA) doubled the yield of colonies. Irradiation of the cells with 3000 rad eliminated their ability to form erythroid bursts, but did not impair the ability of PHA-treated cells to enhance burst formation when added to a fresh batch of cells. This was due to a humoral factor, since media conditioned by PHA-treated washed cells were as effective as the cells themselves. When cells were separated into subpopulations by an adherence procedure and according to their ability to form rosettes with sheep red blood cells, it was found that the PHA-dependent burst-promoting activity released into the medium originated in a nonadherent, nonrosetting (T-cell depleted) cell population.

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MNC-RED A Chemically-Defined Method to Produce Enucleated Red Blood Cells from Adult Peripheral Blood Mononuclear Cells

10.1101/616755 ◽

2019 ◽

Author(s):

Shouping Zhang ◽

Emmanuel N Olivier ◽

Zi Yan ◽

Sandra Suzuka ◽

Karl Roberts ◽

...

Keyword(s):

Cell Culture ◽

Red Blood Cells ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Blood Cells ◽

Mononuclear Cells ◽

Iron Chelator ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells ◽

The Many

AbstractMany methods have been developed to produce red blood cellsin vitrobut translational applications have been hampered by the high cost of production. We have developed R6, a chemically-defined, albumin-free, low-transferrin culture medium, and MNC-RED, a protocol to differentiate peripheral blood mononuclear cells into enucleated erythroid cells that does not require any albumin or any animal components. Erythropoiesis requires large amounts of iron for hemoglobin synthesis. In all existing protocols, these large iron needs are met by increasing the concentration of holo-transferrin. This is necessary because transferrin recycling does not take place in existing erythroid culture conditions. In the R6 medium, iron is provided to the differentiating erythroblasts by small amounts of recombinant transferrin supplemented with FeIII-EDTA, an iron chelator that allows transferrin recycling to take place in cell culture. As a result of the absence of albumin and the use of low amounts of transferrin, the production of cultured red blood cells using the MNC-RED protocol is much less expensive than with existing protocols. The MNC-RED protocol should therefore help make the many translational applications of cultured RBCs economically more feasible.HighlightsWe have developed R6, a chemically-defined, albumin-free low-transferrin culture medium, and MNC-RED, a protocol to differentiate peripheral blood mononuclear cells into enucleated erythroid ER6 is suitable for red blood cell culture despite the low transferrin amounts because of the presence of FeIII-EDTA, an iron chelator that allows transferrin recycling to take place in cell culture.The MNC-RED protocol should help make the many translational applications of cultured RBCs more economically feasible.

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Human erythroid burst-promoting activity produced by phytohemagglutinin- stimulated, radioresistant peripheral blood mononuclear cells

Blood ◽

10.1182/blood.v54.5.1050.bloodjournal5451050 ◽

1979 ◽

Vol 54 (5) ◽

pp. 1050-1057

Author(s):

D Meytes ◽

JA Ma Ortega ◽

NA Shore ◽

PP Dukes

Keyword(s):

T Cell ◽

Red Blood Cells ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Blood Cells ◽

Colony Formation ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

The regulation of erythroid burst-colony formation was studied in cultures of human peripheral blood mononuclear cells. Numbers of erythropoietin-stimulated colonies obtainable from the cells in response to various treatments were compared. One-day preincubation of the cells with phytohemagglutinin (PHA) doubled the yield of colonies. Irradiation of the cells with 3000 rad eliminated their ability to form erythroid bursts, but did not impair the ability of PHA-treated cells to enhance burst formation when added to a fresh batch of cells. This was due to a humoral factor, since media conditioned by PHA-treated washed cells were as effective as the cells themselves. When cells were separated into subpopulations by an adherence procedure and according to their ability to form rosettes with sheep red blood cells, it was found that the PHA-dependent burst-promoting activity released into the medium originated in a nonadherent, nonrosetting (T-cell depleted) cell population.

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Large-scalein-vitroproduction of red blood cells from human peripheral blood mononuclear cells

10.1101/659862 ◽

2019 ◽

Cited By ~ 1

Author(s):

Steven Heshusius ◽

Esther Heideveld ◽

Patrick Burger ◽

Marijke Thiel-Valkhof ◽

Erica Sellink ◽

...

Keyword(s):

Red Blood Cells ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Blood Cells ◽

Mononuclear Cells ◽

Cellular Therapy ◽

Small Scale ◽

Blood Group Antigens ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

AbstractTransfusion of donor-derived red blood cells is the most common form of cellular therapy. Donor availability and the potential risk of alloimmunization and other transfusion-related complications may, however, limit the availability of transfusion units especially for chronically transfused patients.In-vitrocultured, customizable red blood cells would negate these concerns and introduce precision medicine. Large-scale, cost effective production depends on optimization of culture conditions. We developed a defined medium and adapted our protocols to GMP culture requirements, which reproducibly provided pure erythroid cultures from peripheral blood mononuclear cells without prior CD34+isolation, and a 3×107-fold increase in erythroblasts in 25 days. Expanded erythroblast cultures could be differentiated to CD71dimCD235a+CD44+CD117−DRAQ5−red blood cells in 12 days. More than 90% of the cells enucleated and expressed adult hemoglobin as well as the correct blood group antigens. Deformability and oxygen binding capacity of cultured red blood cells was comparable toin-vivoreticulocytes. Daily RNA sampling during differentiation followed by RNA-seq provided a high-resolution map/resource of changes occurring during terminal erythropoiesis. The culture process was compatible with upscaling using a G-Rex bioreactor with a capacity of 1L per reactor, allowing transition towards clinical studies and small-scale applications.

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Tenofovir Diphosphate and Emtricitabine Triphosphate Concentrations in Blood Cells Compared With Isolated Peripheral Blood Mononuclear Cells

JAIDS Journal of Acquired Immune Deficiency Syndromes ◽

10.1097/qai.0b013e3182794723 ◽

2013 ◽

Vol 62 (3) ◽

pp. 260-266 ◽

Cited By ~ 23

Author(s):

Jessica L. Adams ◽

Craig Sykes ◽

Prema Menezes ◽

Heather M.A. Prince ◽

Kristine B. Patterson ◽

...

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Blood Cells ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells ◽

Tenofovir Diphosphate

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Comprehensive Evaluation of the Effects of Long-term Cryopreservation on Peripheral Blood Mononuclear Cells using Flow Cytometry

10.21203/rs.3.rs-1059931/v1 ◽

2021 ◽

Author(s):

Bo Li ◽

Chunmei Yang ◽

Gui Ja ◽

Yansheng Liu ◽

Na Wang ◽

...

Keyword(s):

Flow Cytometry ◽

T Cells ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Effective Utilization ◽

Blood Mononuclear Cells ◽

Important Evidence

Abstract Human peripheral blood mononuclear cells (PBMCs) originate from hematopoietic stem cells (HSCs) in the bone marrow, which mainly includes lymphocytes (T cells, B cells, and natural killer [NK] cells) and monocytes. Cryopreserved PBMCs providing biobank resources are crucial for clinical application or scientific research. Here, we used flow cytometry to explore the influence of long-term cryopreservation on the quality of PBMCs with the aim of providing important evidence for the effective utilization of biobank resources. The PBMCs were isolated from the peripheral blood, which was collected from volunteers in the hospital. After long-term cryopreservation in liquid nitrogen, we analyzed the changes in cell numbers, viability, and multiple subtypes of PBMCs and studied the apoptosis, proliferation, activation, function, and status of T cells in comparison with freshly isolated PBMCs by flow cytometry, and then further tracked the effects of long-term cryopreservation on the same sample. Although the different cell types in the PBMCs dynamically changed compared with those in the freshly isolated samples, PBMC recovery and viability remained stable after long-term cryopreservation, and the number of most innate immune cells (e.g., monocytes and B cells) was significantly reduced compared to that of the freshly isolated PBMCs or long-term cryopreserved PBMCs; more importantly, the proportion of T cell subtypes, apoptosis, proliferation, and functional T cells, except for Tregs, were not affected by long-term cryopreservation. However, the proportions of activated T, naïve T, central memory T, effector T, and effector memory T cells dynamically changed after long-term cryopreservation. This article provides important evidence for the effective utilization of biobank resources. Long-term cryopreserved PBMCs can be partly used as biological resources for clinical research or basic studies, but the effect of cryopreservation on PBMCs should be considered when selecting cell samples, especially in research relating to activating or inhibiting function.

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CD14+ peripheral blood mononuclear cells from chronic myeloid leukemia and normal donors are inhibitory to short- and long-term cultured colony-forming cells

Leukemia Research ◽

10.1016/s0145-2126(99)00171-x ◽

2000 ◽

Vol 24 (3) ◽

pp. 217-231 ◽

Cited By ~ 2

Author(s):

Beate Heissig ◽

Günther Pasternak ◽

Susanne Hörner ◽

Rainer Schwerdtfeger ◽

Siegfried Rossol ◽

...

Keyword(s):

Chronic Myeloid Leukemia ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Myeloid Leukemia ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells ◽

Short And Long Term

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Persistence and Dissemination of Simian Retrovirus Type 2 DNA in Relation to Viremia, Seroresponse, and Experimental Transmissibility in Macaca fascicularis

Journal of Virology ◽

10.1128/jvi.77.20.10751-10759.2003 ◽

2003 ◽

Vol 77 (20) ◽

pp. 10751-10759 ◽

Cited By ~ 11

Author(s):

Roseanne C. Wilkinson ◽

Claire K. Murrell ◽

Rebecca Guy ◽

Gail Davis ◽

Joanna M. Hall ◽

...

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Macaca Fascicularis ◽

Mononuclear Cells ◽

Clinical Signs ◽

Viral Dna ◽

Peripheral Blood Mononuclear ◽

Serological Status ◽

Blood Mononuclear Cells

ABSTRACT Endemic simian retrovirus (SRV) infection can cause fatal simian AIDS in Macaca fascicularis, but many individuals survive with few clinical signs. To further clarify the parameters of SRV pathogenesis, we investigated the persistence of viral DNA forms in relation to active viremia, antibody response, and transmissibility of infection. In M. fascicularis from endemically SRV-2-infected colonies, viral DNA was present in both linear and unintegrated long terminal repeat circular forms in peripheral blood mononuclear cells of all viremic and many nonviremic animals. Long-term followup of three individuals with distinct infection patterns demonstrated persistence of linear and circular forms of viral DNA in peripheral blood mononuclear cells and tissues, irrespective of viremia or antibody status, but reactivation of latent infections was not observed. The role of viral DNA in transmission and early pathogenesis of SRV-2 was investigated by inoculation of SRV-2 DNA-positive blood into groups of naïve M. fascicularis from either a viremic or nonviremic donor and subsequent analysis of the virological and serological status of the recipients. Transmission of SRV and development of anti-SRV antibodies were only observed in recipients of blood from the viremic donor; transfer of SRV provirus and unintegrated circular DNA in blood from the nonviremic donor did not lead to infection of the recipients. These results indicate that a proportion of M. fascicularis are able to effectively control the replication and infectivity of SRV despite long-term persistence of viral DNA forms in infected lymphocytes.

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