Analysis of Mitochondrial Dimensions and Cristae Structure in Pluripotent Stem Cells Using Transmission Electron Microscopy

2018 ◽  
Vol 47 (1) ◽  
pp. e67 ◽  
Author(s):  
I. C. Tobias ◽  
R. Khazaee ◽  
D. H. Betts
Keyword(s):  
Electron Microscopy ◽  
Stem Cells ◽  
Transmission Electron Microscopy ◽  
Pluripotent Stem Cells ◽  
Transmission Electron

scholarly journals Satellite Cells CD44 Positive Drive Muscle Regeneration in Osteoarthritis Patients

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Manuel Scimeca ◽  
Elena Bonanno ◽  
Eleonora Piccirilli ◽  
Jacopo Baldi ◽  
Alessandro Mauriello ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Stem Cells ◽  
Transmission Electron Microscopy ◽  
Satellite Cells ◽  
Muscle Regeneration ◽  
Bone Diseases ◽  
Mineral Density ◽  
Muscle Stem Cells ◽  
Age Related ◽  
Transmission Electron

Age-related bone diseases, such as osteoarthritis and osteoporosis, are strongly associated with sarcopenia and muscle fiber atrophy. In this study, we analyzed muscle biopsies in order to demonstrate that, in osteoarthritis patients, both osteophytes formation and regenerative properties of muscle stem cells are related to the same factors. In particular, thanks to immunohistochemistry, transmission electron microscopy, and immunogold labeling we investigated the role of BMP-2 in muscle stem cells activity. In patients with osteoarthritis both immunohistochemistry and transmission electron microscopy allowed us to note a higher number of CD44 positive satellite muscle cells forming syncytium. Moreover, the perinuclear and cytoplasmic expression of BMP-2 assessed byin situmolecular characterization of satellite cells syncytia suggest a very strict correlation between BMP-2 expression and muscle regeneration capability. Summing up, the higher BMP-2 expression in osteoarthritic patients could explain the increased bone mineral density as well as decreased muscle atrophy in osteoarthrosic patients. In conclusion, our results suggest that the control of physiological BMP-2 balance between bone and muscle tissues may be considered as a potential pharmacological target in bone-muscle related pathology.

Modeling Congenital Dyserythropoietic Anemia Type I Through Patient-Derived Induced Pluripotent Stem Cells

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 3196-3196
Author(s):  
Orna Steinberg Shemer ◽  
Yu Yao ◽  
Gary M. Kupfer ◽  
Hannah Tamary ◽  
Mitchell J. Weiss
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Light Microscopy ◽  
Chromatin Structure ◽  
Pluripotent Stem Cells ◽  
Type I ◽  
Transmission Electron ◽  
Erythroid Precursors ◽  
Induced Pluripotent ◽  
I Cells

Abstract Abstract 3196 Congenital dyserythropoietic anemias (CDAs) are rare inherited disorders characterized by impaired red blood cell formation (dyserythropoiesis) and signature cytopathologies. CDA type I is an autosomal recessive disease with macrocytic anemia and occasional bone abnormalities. Erythroid precursors exhibit pathognomonic abnormalities including internuclear chromatin bridges and spongy (“Swiss cheese”) heterochromatin. The disease is caused by biallelic mutations in the gene CDANI (Dgany et al., 2002), which encodes codanin-1, a ubiquitously expressed protein that is believed to have fundamental roles in cell cycle control and chromatin structure (Noy-Lotan et. al, 2009). Animal models for the study of CDA I are suboptimal and clinical samples are scarce. Thus, we have developed an experimental model for the study of CDA I by generating induced pluripotent stem cells (iPSCs) from affected patients. We reprogrammed fibroblasts from CDA I patients and normal subjects using a single lentiviral vector encoding OCT4, KLF4, SOX2, and MYC. The resultant iPSCs exhibited standard criteria for pluripotency and the integrated reprogramming vector was excised using Cre-lox technology. We differentiated CDA I and control iPSCs into erythroid progenitors by inducing the formation of embryoid bodies (EBs) with stepwise additions of supportive cytokines. Beginning at about day 8, hematopoietic progenitors with erythroid potential were detected within EBs and as free-floating cells in the medium. Our differentiation protocol showed two waves of erythroid precursor production. Early EBs (days 12 to 23) produced erythroid cells that expressed mainly epsilon globin, resembling early yolk sac type “primitive” erythropoiesis. In contrast, erythroblasts produced from later EBs (days 27 to 50) expressed mainly gamma globins, resembling “definitive” erythroid cells produced by late stage yolk sac and fetal liver. Our preliminary studies, indicate that CDA I iPSCs produce normal numbers of primitive and definitive erythrocytes. No defects in survival or maturation were detected by flow cytometry assessing the expression of annexin V and the developmental stage markers CD235/CD71/forward scatter. However, definitive type (but not primitive) erythroblasts derived from CDA I iPSCs exhibit some characteristic pathological features including occasional internuclear chromatin bridging visible by light microscopy and spongy “Swiss cheese” heterochromatin revealed by transmission electron microscopy. Thus, patient-derived iPSCs can model at least some aspects of CDA I and provide the basis for future studies to define the actions of codanin-1 and the pathophysiology of this disorder. Figure: Patient iPSC-derived erythroblasts recapitulate CDA I pathology. Light microscopy and transmission electron microscopy (TEM) of normal and CDA I iPSC-derived erythroblasts generated in ∼30 day differentiation cultures. Inserts show higher magnification of the marked areas. CDA I cells exhibit occasional internuclear bridges on light microscopy (third panel). TEM showed abnormal spongy chromatin structure in most CDA I erythroid precursors (fourth panel). Figure:. Patient iPSC-derived erythroblasts recapitulate CDA I pathology. Light microscopy and transmission electron microscopy (TEM) of normal and CDA I iPSC-derived erythroblasts generated in ∼30 day differentiation cultures. Inserts show higher magnification of the marked areas. CDA I cells exhibit occasional internuclear bridges on light microscopy (third panel). TEM showed abnormal spongy chromatin structure in most CDA I erythroid precursors (fourth panel). Disclosures: No relevant conflicts of interest to declare.

scholarly journals Influence of surface-modified maghemite nanoparticles on in vitro survival of human stem cells

2014 ◽  
Vol 5 ◽  
pp. 1732-1737 ◽  
Author(s):  
Michal Babič ◽  
Daniel Horák ◽  
Lyubov L Lukash ◽  
Tetiana A Ruban ◽  
Yurii N Kolomiets ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Stem Cells ◽  
Transmission Electron Microscopy ◽  
Precipitation Method ◽  
Maghemite Nanoparticles ◽  
Human Stem Cells ◽  
Transmission Electron ◽  
Surface Modified ◽  
In Vitro Survival

Surface-modified maghemite (γ-Fe2O3) nanoparticles were obtained by using a conventional precipitation method and coated with D-mannose and poly(N,N-dimethylacrylamide). Both the initial and the modified particles were characterized by transmission electron microscopy and dynamic light scattering with regard to morphology, particle size and polydispersity. In vitro survival of human stem cells was then investigated by using the methyl thiazolyl tetrazolium (MTT) assay, which showed that D-mannose- and poly(N,N-dimethylacrylamide)-coated γ-Fe2O3 particles exhibit much lower level of cytotoxicity than the non-coated γ-Fe2O3.

scholarly journals Ultrastructural analysis of different human mesenchymal stem cells after in vitro expansion: a technical review

2015 ◽  
Vol 59 (4) ◽  
Author(s):  
M. Miko ◽  
L. Danisovic ◽  
A. Majidi ◽  
I. Varga
Keyword(s):  
Electron Microscopy ◽  
Stem Cells ◽  
Transmission Electron Microscopy ◽  
Mesenchymal Stem Cells ◽  
Regenerative Medicine ◽  
Adult Stem Cells ◽  
Cell Populations ◽  
Histological Techniques ◽  
Transmission Electron

Transmission electron microscopy reveals ultrastructural details of cells, and it is a valuable method for studying cell organelles. That is why we used this method for detailed morphological description of different adult tissuederived stem cells, focusing on the morphological signs of their functions (proteosynthetic activity, exchange with external environment, <em>etc</em>.) and their comparison. Preparing a specimen from the cell culture suitable for transmission electron microscopy is, however, much more challenging than routine tissue processing for normal histological examination. There are several issues that need to be solved while working with cell pellets instead of solid tissue. Here we describe a simple protocol for the isolation and culture of mesenchymal stem cells from different adult tissues, with applications to stem cell biology and regenerative medicine. Since we are working with population of cells that was obtained after many days of passaging, very efficient and gentle procedures are highly necessary. We demonstrated that our semi-conservative approach regarding to histological techniques and processing of cells for transmission electron microscopy is a well reproducible procedure which results in quality pictures and images of cell populations with minimum distortions and artifacts. We also commented about riskiest steps and histochemical issues (<em>e.g</em>., precise pH, temperature) while preparing the specimen. We bring full and detailed procedures of fixation, post-fixation, infiltration, embedding, polymerization and contrasting of cell obtained from <em>in vitro</em> cell and tissue cultures, with modifications according to our experience. All this steps are essential for us to know more about adult stem cells derived from different sources or about other random cell populations. The knowledge about detailed ultra-structure of adult stem cells cultured <em>in vitro</em> are also essential for their using in regenerative medicine and tissue engineering.

Techniques for Transmission Electron Microscopy of Fiber-Matrix Interfaces in Aluminum Alloy-Boron Composites by Ion Erosio

1971 ◽  
Vol 29 ◽  
pp. 204-205
Author(s):  
G. G. Shaw
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Aluminum Alloy ◽  
Electron Beam ◽  
Pure Aluminum ◽  
Ion Milling ◽  
Transmission Electron ◽  
Fiber Matrix ◽  
Ion Erosion ◽  
Row Of Fibers

The morphology and composition of the fiber-matrix interface can best be studied by transmission electron microscopy and electron diffraction. For some composites satisfactory samples can be prepared by electropolishing. For others such as aluminum alloy-boron composites ion erosion is necessary.When one wishes to examine a specimen with the electron beam perpendicular to the fiber, preparation is as follows: A 1/8 in. disk is cut from the sample with a cylindrical tool by spark machining. Thin slices, 5 mils thick, containing one row of fibers, are then, spark-machined from the disk. After spark machining, the slice is carefully polished with diamond paste until the row of fibers is exposed on each side, as shown in Figure 1.In the case where examination is desired with the electron beam parallel to the fiber, preparation is as follows: Experimental composites are usually 50 mils or less in thickness so an auxiliary holder is necessary during ion milling and for easy transfer to the electron microscope. This holder is pure aluminum sheet, 3 mils thick.

Direct Observation of Heat Exchanger Deposits by Cross Sectioning

1967 ◽  
Vol 25 ◽  
pp. 364-365
Author(s):  
R. W. Anderson ◽  
D. L. Senecal
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Heat Exchanger ◽  
Direct Observation ◽  
Cross Sections ◽  
Preparation Method ◽  
Specimen Preparation ◽  
Flowing Water ◽  
Transmission Electron ◽  
Deposit Layer

A problem was presented to observe the packing densities of deposits of sub-micron corrosion product particles. The deposits were 5-100 mils thick and had formed on the inside surfaces of 3/8 inch diameter Zircaloy-2 heat exchanger tubes. The particles were iron oxides deposited from flowing water and consequently were only weakly bonded. Particular care was required during handling to preserve the original formations of the deposits. The specimen preparation method described below allowed direct observation of cross sections of the deposit layers by transmission electron microscopy.The specimens were short sections of the tubes (about 3 inches long) that were carefully cut from the systems. The insides of the tube sections were first coated with a thin layer of a fluid epoxy resin by dipping. This coating served to impregnate the deposit layer as well as to protect the layer if subsequent handling were required.

Phase Transformations in Ti-7w/oMo-3w/oMe Alloy

1974 ◽  
Vol 32 ◽  
pp. 514-515
Author(s):  
S. Fujishiro
Keyword(s):  
Electron Microscopy ◽  
Mechanical Properties ◽  
Transmission Electron Microscopy ◽  
Tensile Strength ◽  
Mechanical Processing ◽  
Ray Method ◽  
X Ray ◽  
Transmission Electron ◽  
Water Quench ◽  
Alpha Beta

The mechanical properties of three titanium alloys (Ti-7Mo-3Al, Ti-7Mo- 3Cu and Ti-7Mo-3Ta) were evaluated as function of: 1) Solutionizing in the beta field and aging, 2) Thermal Mechanical Processing in the beta field and aging, 3) Solutionizing in the alpha + beta field and aging. The samples were isothermally aged in the temperature range 300° to 700*C for 4 to 24 hours, followed by a water quench. Transmission electron microscopy and X-ray method were used to identify the phase formed. All three alloys solutionized at 1050°C (beta field) transformed to martensitic alpha (alpha prime) upon being water quenched. Despite this heavily strained alpha prime, which is characterized by microtwins the tensile strength of the as-quenched alloys is relatively low and the elongation is as high as 30%.

Two- or three-way observations of the identical cell elements within the same sections by LM, TEM and/or SEM

1978 ◽  
Vol 36 (2) ◽  
pp. 82-83 ◽  
Author(s):  
Nakazo Watari ◽  
Yasuaki Hotta ◽  
Yoshio Mabuchi
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fine Structure ◽  
Light Microscopy ◽  
Thin Sections ◽  
Transmission Electron ◽  
Identical Cell ◽  
Electron Microscopes ◽  
Scanning Electron

It is very useful if we can observe the identical cell elements within the same sections by light microscopy (LM), transmission electron microscopy (TEM) and/or scanning electron microscopy (SEM) sequentially, because, the cell fine structure can not be indicated by LM, while the color is; on the other hand, the cell fine structure can be very easily observed by EM, although its color properties may not. However, there is one problem in that LM requires thick sections of over 1 μm, while EM needs very thin sections of under 100 nm. Recently, we have developed a new method to observe the same cell elements within the same plastic sections using both light and transmission (conventional or high-voltage) electron microscopes.In this paper, we have developed two new observation methods for the identical cell elements within the same sections, both plastic-embedded and paraffin-embedded, using light microscopy, transmission electron microscopy and/or scanning electron microscopy (Fig. 1).

Comparative Microstructures of Ion Milled and Electrochemically Thinned Composite Materials

1974 ◽  
Vol 32 ◽  
pp. 546-547
Author(s):  
R.R. Russell
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Composite Materials ◽  
Great Difficulty ◽  
Ion Milling ◽  
Transmission Electron ◽  
Ion Damage ◽  
Intermetallic Composite

Transmission electron microscopy of metallic/intermetallic composite materials is most challenging since the microscopist typically has great difficulty preparing specimens with uniform electron thin areas in adjacent phases. The application of ion milling for thinning foils from such materials has been quite effective. Although composite specimens prepared by ion milling have yielded much microstructural information, this technique has some inherent drawbacks such as the possible generation of ion damage near sample surfaces.

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