Evidence of marked glycogen variations in the characteristic Raman signatures of human embryonic stem cells

Human embryonic stem cells (hESCs) have typical Raman signatures, but specific factors that contribute to variations in these signatures have not been reported to date. Furthermore, variations due to the passaging that is necessary for hESC culture maintenance could potentially distort these signatures. It is therefore important to characterize the impact of these culture manipulations on the Raman spectra to gain a better understanding of the origins and nature of their variations. Here we report on the Raman microspectroscopy of hESCs samples from maintenance cultures, complemented with periodic acid Schiff (PAS, carbohydrates) and 4′‐6‐diamidino‐2‐phenlyindol (DAPI, nuclei) staining. The component predominantly responsible for variations between spectra was spectrally identified as glycogen. Variations in the Raman map of the 480 cm⁻¹ glycogen marker band corresponded with those of a PAS stain of the same sample area. The 785‐nm Raman microspectra of hESC cultures examined daily after passaging showed that the same nonrandom spectral variances occurred at all time points after passaging. The pattern of these variances was identified as being due to glycogen spectral components. Our results help validate the previously observed spectral signatures of hESCs and further delineate and characterize the variations that can be expected in these signatures under normal maintenance culture conditions, and aid distinguishing them from those corresponding to differentiation, thus providing a benchmark for future studies. Copyright © 2010 John Wiley & Sons, Ltd.     

Solid Phase Extraction of Lead(II) by Sorption on Grinded Eucalyptus Stem and Determination with Flame Atomic Absorption Spectrometry

A sensitive and simple solid‐phase preconcentration procedure for the determination of trace amount of lead by flame atomic absorption spectrometry (FAAS) is developed. The method is based on the adsorption of Pb²⁺ on the column of fine grinded eucalyptus stem adsorbent, elution of the column by nitric acid and subsequent determination by FAAS. The effect of different variables such as pH, eluent type, flow rate and interfering ions on the recovery of the analyte was investigated and optimum conditions were established. The adsorption of lead onto fine grinded eucalyptus stem can formally be described by a Langmuir equation with a maximum adsorption capacity of 4.49 mg g^?1. A preconcentration factor of 50 was achieved using the optimum conditions. The calibration graph was linear in the range 10–125 ng mL^?1 of lead in the initial solution with r = 0.9982. The limit of detection based on 3S_b criterion was 4.5 ng mL^?1 and the relative standard deviation for eight replicate measurements of 30 and 80 ng mL^?1 of iron was 3.6 and 2.8%, respectively. The method was successfully applied to the determination of lead added to well, tap and wastewater samples.     

Thromboxane A2 modulates migration, proliferation, and differentiation of adipose tissue-derived mesenchymal stem cells

Prostanoid metabolites are key mediators in inflammatory responses, and accumulating evidence suggests that mesenchymal stem cells (MSCs) can be recruited to injured or inflamed tissues. In the present study, we investigated whether prostanoid metabolites can regulate migration, proliferation, and differentiation potentials of MSCs. We demonstrated herein that the stable thromboxane A₂ (TxA₂) mimetic U46619 strongly stimulated migration and proliferation of human adipose tissue-derived MSCs (hADSCs). Furthermore, U46619 treatment increased expression of α-smooth muscle actin (α-SMA), a smooth muscle marker, in hADSCs, suggesting differentiation of hADSCs into smooth muscle-like cells. U46619 activated ERK and p38 MAPK, and pretreatment of the cells with the MEK inhibitor U0126 or the p38 MAPK inhibitor SB202190 abrogated the U46619-induced migration, proliferation, and α-SMA expression. These results suggest that TxA₂ plays a key role in the migration, proliferation, and differentiation of hADSCs into smooth muscle-like cells through signaling mechanisms involving ERK and p38 MAPK.     

Change in serum proteome during allogeneic hematopoietic stem cell transplantation and clinical significance of serum C-reactive protein and haptoglobin

Successful hematopoietic stem cell transplantation (HSCT) involves the restoration of hematopoietic function after engraftment, arising from the differentiation and proliferation of hematopoietic stem cells. Several factors could influence the course of allogeneic-HSCT (allo-HSCT). Therefore, knowledge of serum proteome changes during the allo-HSCT period might increase the efficacy of diagnosis and disease prevention efforts. This study conducted proteomic analyses to find proteins that were significantly altered in response to allo-HSCT. Sera from five representative patients who underwent allo-HSCT were analyzed by 2-dimensional gel electrophoresis and liquid chromatography tandem mass spectrometry, and were measured on a weekly basis before and after allo-HSCT in additional 78 patients. Fourteen protein spots showing changes in expression were further examined, and most proteins were identified as acute phase proteins (APPs). Studies of 78 additional patients confirmed that C-reactive protein (CRP) and haptoglobin undergo expression changes during allo-HSCT and thus may have the potential to serve as representative markers of clinical events after allo-HSCT. Maximal CRP level affected the development of major transplant-related complications (MTCs) and other problems such as fever of unknown origin. Particularly, an increase in CRP level 21 days after allo-HSCT was found to be an independent risk factor for MTC. Maximal haptoglobin and haptoglobin level 14 days after allo-HSCT were predictive of relapses in underlying hematologic disease. Our results indicated that CRP and haptoglobin were significantly expressed during allo-HSCT, and suggest that their level can be monitored after allo-HSCT to assess the risks of early transplant-related complications and relapse.     

Thick Soft Tissue Reconstruction on Highly Perfusive Biodegradable Scaffolds

The lack of a vascular network and poor perfusion is what mostly prevents three‐dimensional (3D) scaffolds from being used in organ repair when reconstruction of thick tissues is needed. Highly‐porous scaffolds made of poly(L ‐lactic acid) (PLLA) are prepared by directional thermally induced phase separation (dTIPS) starting from 1,4‐dioxane/PLLA solutions. The influence of polymer concentration and temperature gradient, in terms of imposed intensity and direction, on pore size and distribution is studied by comparison with scaffolds prepared by isotropic TIPS. The processing parameters are optimized to achieve an overall porosity for the 3D scaffolds of about 93% with a degree of interconnectivity of 91%. The resulting pore network is characterized by the ordered repetition of closely packed dendrite‐like cavities, each one showing stacks of 20 µm large side lamellar branches departing from 70 µm diameter vertical backbones, strongly resembling the vascular patterns. The in vitro biological responses after 1 and 2 weeks are evaluated from mesenchymal (bone marrow stromal) cells (MSC) static culturing. A novel vacuum‐based deep‐seeding method is set up to improve uniform cell penetration down to scaffold thicknesses of over 1 mm. Biological screenings show significant 3D scaffold colonization even after 18 h, while cellular retention is observed up to 14 d in vitro (DIV). Pore architecture‐driven cellular growth is accompanied by cell tendency to preserve their multi‐potency towards differentiation. Confluent tissues as thick as 1 mm were reconstructed taking advantage of the large perfusion enhanced by the highly porous microstructure of the engineered scaffolds, which could successfully serve for applications aimed at vascular nets and angiogenesis.

     

Control of Osteogenic Differentiation and Mineralization of Human Mesenchymal Stem Cells on Composite Nanofibers Containing Poly[lactic‐co‐(glycolic acid)] and Hydroxyapatite

We fabricated composite fibrous scaffolds from blends of poly(lactide‐co‐glycolide) (PLGA) and nano‐sized hydroxyapatite (HA) via electrospinning. SEM‐EDX and AFM analysis demonstrated that HA was homogeneously dispersed in the nanofibers, and the roughness increased along with the amount of incorporated HA. When hMSCs were cultured on these PLGA/HA composite nanofibers, we found that incorporation of HA on the nanofibers did not affect cell viability whereas increased ALP activity and expression of osteogenic genes as well as the calcium mineralization of hMSCs. Our results indicate that the composite nanofibers can be offered as a potential bone regenerative biomaterial for stem cell based therapies.

     

小麦茎秆性状与单穗重的相关分析与通径分析

利用2010-2011连续两年河南科技学院小麦试验田所得的试验资料,对6个不同小麦品种的茎秆性状与单穗重进行相关分析与通径分析.相关分析结果表明,所考查的茎秆性状与单穗重的相关程度依次为:茎重>茎粗>株高>茎长>壁厚;茎重与单穗重(r_(5y)=0.8213~(**))和茎粗与茎重(r_(25)=0.7218~(**))、茎长与株高(r_(13)=0.9582~(**))的相关分别呈极显著,茎粗与单穗重(r_(2y)=0.5990~*)、茎重与株高(r_(13)=0.6550~*)均达显著水平.通径分析结果显示,考查的茎部性状对单穗重的直接效应(绝对值)依次为:茎重>株高>茎长>壁厚>茎粗.茎重的直接正效应(p_(x_5→x_5→y)=1.3986~(**))最大,茎长(p_(x_1→x_1→y)=0.6297~*)次之,壁厚(p_(x_3→x_3→y)=0.2323)居三,株高的直接负效应(p_(x_3→x_3→y)=-1.1617~(**))极显著;5个茎部性状的决策系数依次为:茎重(0.3414)>壁厚(-0.0030)>茎粗(-0.1959)>茎长(一0.2063)>株高(一1.9800).据此明确了茎重、壁厚为提高小麦单穗重的主选因素,株高是主控因素.在注重对基部节间短、粗、茎壁厚性状选择的同时,适当控制株高,是选育高产抗倒小麦品种的有效途径.     

Human Elastin‐Based Recombinant Biopolymers Improve Mesenchymal Stem Cell Differentiation

Elastin‐based polypeptides are a class of smart biopolymers representing an important model in the design of biomaterials. The combination of biomimetic materials with cells that have great plasticity provides a promising strategy for the realization of highly engineered cell‐based constructs for regenerative medicine and tissue repair applications. Two recombinant biopolymers inspired by human elastin are assessed as coating agents to prepare biomimetic surfaces for cell culture. These substrates are assayed for hBM MSC culture. The coated surfaces are also characterized with AFM to evaluate the topographical features of the deposited biopolymers. The results suggest that the elastin‐derived biomimetic surfaces play a stimulatory role on osteogenic differentiation of MSCs.

     

Viability of Human Mesenchymal Stem Cells Seeded on Crosslinked Entropy‐Elastic Gelatin‐Based Hydrogels

Biomimetic polymer network systems with tailorable properties based on biopolymers represent a class of degradable hydrogels that provides sequences for protein adsorption and cell adhesion. Such materials show potential for in vitro MSC proliferation as well as in vivo applications and were obtained by crosslinking different concentrations of gelatin using varying amounts of ethyl lysine diisocyanate in the presence of a surfactant in pH 7.4 PBS solution. Material extracts, which were tested for cytotoxic effects using L929 mouse fibroblasts, were non‐toxic. The hydrogels were seeded with human bone marrow‐derived MSCs and supported viable MSCs for the incubation time of 9 d. Preadsorption of fibronectin on materials improved this biofunctionality.