Red blood cells (RBCs) and platelets derived from stem cells are possible solutions to the increasing demand for blood transfusion. Based on the availability of stem cells, their relatively defined ...differentiation mechanisms, and the massive exploration of induction systems, the generation of RBCs or platelets in vitro from cord blood hematopoietic stem/progenitor cells (CB‐HSPCs) has potential for clinical applications. However, information on the clinical translation of stem cell‐derived RBCs and platelets in the literature and at the ClinicalTrials.gov website is very limited. The only clinical trial on cultured RBCs, which aimed to assess the lifespan of RBCs cultured in vivo, was reported by Luc Douay and colleagues. Of note, the cultured RBCs they used were derived from autologous peripheral blood HSPCs, and no cultured platelets have been applied clinically to date. However, CB‐HSPC‐derived megakaryocytes, platelet precursors, have been used in the treatment of thrombocytopenia. A successful phase I trial was reported, followed by phase II and III clinical trials conducted in China. In this review, the gap between the many basic studies and limited clinical trials on stem cell‐derived RBCs and platelets is summarized. The possible reasons and solutions for this gap are discussed. Further technological improvements for blood cell expansion and maturation ex vivo and the establishment of biological standards for stem cell derivatives might help to facilitate the therapeutic applications of cultured RBCs and platelets derived from CB‐HSPCs in the near future.
We review the progress in producing red blood cells (RBCs) and platelets from human cord blood hematopoietic stem/progenitor cells (HSPCs) and relatively limited reports on the clinical trials with cultured RBCs and megakaryocytic progenitors (MPs). Technical improvement together with the establishment of biological standards for stem cell derivations might help to achieve therapeutic applications of cultured RBCs and platelets. EPCs, erythroid progenitor cells.
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FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
Erythroblast enucleation is a precisely regulated but not clearly understood process. Polycythemia shows pathological erythroblast enucleation, and we discovered a low miR-125b-5p level in terminal ...erythroblasts of patients with polycythemia vera (PV) compared to those of healthy controls. Exogenous upregulation of miR-125b-5p levels restored the enucleation rate to normal levels. Direct downregulation of miR-125b-5p in mouse erythroblasts simulated the enucleation issue found in patients with PV, and miR-125b-5p accumulation was found in enucleating erythroblasts, collectively suggesting the importance of miR-125b-5p accumulation for erythroblast enucleation. To elucidate the role of miR-125b-5p in enucleation, gain- and loss-of-function studies were performed. Overexpression of miR-125b-5p improved the enucleation of erythroleukemia cells and primary erythroblasts. Infused erythroblasts with higher levels of miR-125b-5p also exhibited accelerated enucleation. In contrast, miR-125b-5p inhibitors significantly suppressed erythrocyte enucleation. Intracellular imaging revealed that in addition to cytoskeletal assembly and nuclear condensation, miR-125b-5p overexpression resulted in mitochondrial reduction and depolarization. Real-time PCR, western blot analysis, luciferase reporter assays, small molecule inhibitor supplementation and gene rescue assays revealed that Bcl-2, as a direct target of miR-125b-5p, was one of the key mediators of miR-125b-5p during enucleation. Following suppression of Bcl-2, the activation of caspase-3 and subsequent activation of ROCK-1 resulted in cytoskeletal rearrangement and enucleation. In conclusion, this study is the first to reveal the pivotal role of miR-125b-5p in erythroblast enucleation.
Due to the low number of collectable stem cells from single umbilical cord blood (UCB) unit, their initial uses were limited to pediatric therapies. Clinical applications of UCB hematopoietic stem ...and progenitor cells (HSPCs) would become feasible if there were a culture method that can effectively expand HSPCs while maintaining their self-renewal capacity. In recent years, numerous attempts have been made to expand human UCB HSPCs
in vitro
. In this study, we report that caffeic acid phenethyl ester (CAPE), a small molecule from honeybee extract, can promote
in vitro
expansion of HSPCs. Treatment with CAPE increased the percentage of HSPCs in cultured mononuclear cells. Importantly, culture of CD34
+
HSPCs with CAPE resulted in a significant increase in total colony-forming units and high proliferative potential colony-forming units. Burst-forming unit-erythroid was the mostly affected colony type, which increased more than 3.7-fold in 1 μg mL
−1
CAPE treatment group when compared to the controls. CAPE appears to induce HSPC expansion by upregulating the expression of SCF and HIF1-α. Our data suggest that CAPE may become a potent medium supplement for
in vitro
HSPC expansion.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Understanding how hepatic precursor cells can generate differentiated bile ducts is crucial for studies on epithelial morphogenesis and for development of cell therapies for hepatobiliary diseases. ...Epimorphin (EPM) is a key morphogen for duct morphogenesis in various epithelial organs. The role of EPM in bile duct formation (DF) from hepatic precursor cells, however, is not known. To address this issue, we used WB-F344 rat epithelial stem-like cells as model for bile duct formation. A micropattern and a uniaxial static stretch device was used to investigate the effects of EPM and stress fiber bundles on the mitosis orientation (MO) of WB cells. Immunohistochemistry of liver tissue sections demonstrated high EPM expression around bile ducts in vivo. In vitro, recombinant EPM selectively induced DF through upregulation of CK19 expression and suppression of HNF3alpha and HNF6, with no effects on other hepatocytic genes investigated. Our data provide evidence that EPM guides MO of WB-F344 cells via effects on stress fiber bundles and focal adhesion assembly, as supported by blockade EPM, beta1 integrin, and F-actin assembly. These blockers can also inhibit EPM-induced DF. These results demonstrate a new biophysical action of EPM in bile duct formation, during which determination of MO plays a crucial role.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Wellbore instability and reservoir damage caused by filtrate of water-based drilling fluids (WDFs) invading the formation are extremely problematic issues in the process of oil excavation. Herein, we ...reported a novel polymer with core-shell structure based on modified silica nanoparticles as a filtrate reducer (NS-DA) in WDFs. Fourier transform infrared spectroscopy (FTIR) and proton nuclear magnetic resonance (NMR) results revealed that the target product NS-DA had been gained. The results of thermogravimetric analysis (TGA) showed that the thermal decomposition temperature of NS-DA occurred after 293 °C. Meanwhile, NS-DA exhibited the most outstanding filtration loss reduction ability compared to three commercially filtrate reducer (Driscal D. PAC-Lv and CMC) in the filtration loss test prescribed by the American Petroleum Institute. Specifically, the filtration loss of Driscal D/WDFs, PAC-Lv/WDFs and CMC/WDFs were 16.2 mL, 18.4 mL and 19.8 mL after hot rolling at 180 °C for 16 h, respectively, while the filtration loss of NS-DA/WDFs was only 5.6 mL. The mechanism of NS-DA in controlling filtration loss was investigated by Zeta potential measurements, particle size distribution analysis and mud cake analysis. The results showed that NS-DA molecular chains were firmly adsorbed on the surface of clay particles through hydrogen and ionic bonds, which reduced the zeta potential and the average particle size of clay particles, diminished the mud cake permeability, and thus enhanced the stability of the WDFs colloidal system. This work indicates that NS-DA has great potential for future oil excavation.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The rapid growth of global energy demand necessitates high-performance water-based drilling fluids (WDFs) with excellent-plugging performance in the deep excavation of shale gas formation. Herein, we ...report an organic-inorganic nanocomposite (NS-D) as a plugging agent in WDFs to plug the nanoporous of shale and abate the hydration expansion of shale by integrating the advantages of inorganic and polymer nano plugging agents. The results of Fourier Transform infrared spectroscopy (FTIR) and proton nuclear magnetic resonance (H-NMR) revealed that NS-D had been successfully obtained. Additionally, thermogravimetric analysis (TGA) showed that the thermal degradation temperature of NS-D occurred after 354 °C. The results of plugging theory analysis showed that the presence of rich amide groups and rigid material silica not only made NS-D adhere to shale efficiently by hydrogen bonding, but also firmly blocked the nanoporous of shale. The plugging ability was evaluated by pressure transfer test and nitrogen adsorption test. The results showed that WDFs with NS-D can significantly improve plugging efficiency and reduce fluid invasion. On the other hand, NS-D sharply abated the fluid loss of WDFs due to its positive plugging performance. The filtrate volume of WDFs decreased by 90.5% and 88.3%, respectively, before and after hot rolling at 150°C for 16 h. Moreover, the plugging performance of NS-D for WDFs was significantly better than polymer plugging agent crosslinked polyacrylamide microspheres and inorganic plugging agent silica nanoparticles. This work provides a novel strategy to plug the nanoporous of shale in the process of shale gas exploration.
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BFBNIB, GIS, IJS, KISLJ, NUK, PNG, UL, UM, UPUK
With the development and utilization of offshore liquefied natural gas, it is increasingly important to study the influence of the heat transfer performance of a spiral-wound heat exchanger under ...sloshing conditions. This study focused on the effects of different sloshing amplitudes and sloshing periods on the heat transfer and pressure drop performance of a heat exchanger. Through experimental research, the results showed that the fluctuation of the UA (U is the heat transfer coefficient; A is the heat exchange area) value first increased and then decreased with an increase in the sloshing amplitude. The UA value increased by 12.92% and decreased by 42.03% compared to the static value at 3 and 9°, respectively. The fluctuation in the UA value first decreased and then increased with an increase in the sloshing period. The UA value decreased by 36.66% and increased by 10.82% slowly compared to the static value when the sloshing period was 6 and 20 s, respectively. Based on this, a mathematical model of heat transfer under the condition of pitch sloshing was established.
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IJS, KILJ, NUK, PNG, UL, UM, UPUK
To build a protocol of separation and induction of megakaryocytes derived from cord blood mononuclear cells.
Red blood cells were precipitated by hydroxyethyl starch (HES). Mononuclear cells were ...obtained by density gradient centrifugation with Ficoll. The inducing efficiencies of megakaryocytes by using of different cytokine cocktails and culture media were analyzed.
The best choice for erythrocyte sedimentation and high efficiency of nucleated cells retrieving were obtained by using of 1.5% HES. The isolated cord blood mononuclear cells were cultured with domestic serum-free medium supplemented with 116t (IL-11, IL-6, TPO), st36(SCF, TPO, IL-3, IL-6), pt36 (PDGF,TPO,IL-3,IL-6) or pst36 for 7 days. St36 group (50 ng/ml SCF, 50 ng/ml TPO, 20 ng/ml IL-3 and 50 ng/ml IL-6) yielded the most CD41/CD61 positive (6.79±1.97)×10⁴. The cell viability (82.85 ± 0.64)% of st36 group by using of imported serum-free medium was better than (60.90±6.93)% that in domestic medium on day 7 after induction, and CD41/CD61 p
Cell-cell contacts and interactions between pancreatic β-cells and/or other cell populations within islets are essential for cell survival, insulin secretion, and functional synchronization. ...Three-dimensional (3D) culture systems supply the ideal microenvironment for islet-like cluster formation and functional maintenance. However, the underlying mechanisms remain unclear. In this study, mouse insulinoma 6 (MIN6) cells were cultured in a rotating 3D culture system to form islet-like aggregates. Glucose-stimulated insulin secretion (GSIS) and the RhoA/ROCK pathway were investigated. In the 3D-cultured MIN6 cells, more endocrine-specific genes were up-regulated, and GSIS was increased to a greater extent than in cells grown in monolayers. RhoA/ROCK inactivation led to F-actin remodeling in the MIN6 cell aggregates and greater insulin exocytosis. The gap junction protein, connexin 36 (Cx36), was up-regulated in MIN6 cell aggregates and RhoA/ROCK-inactivated monolayer cells. GSIS dramatically decreased when Cx36 was knocked down by short interfering RNA and could not be reversed by RhoA/ROCK inactivation. Thus, the RhoA/ROCK signaling pathway is involved in insulin release through the up-regulation of Cx36 expression in 3D-cultured MIN6 cells.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OBVAL, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
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