Time-lapse microscopy and classification of 2D human mesenchymal stem cells based on cell shape picks up myogenic from osteogenic and adipogenic differentiation

Current methods to characterize mesenchymal stem cells (MSCs) are limited to CD marker expression, plastic adherence and their ability to differentiate into adipogenic, osteogenic and chondrogenic precursors. It seems evident that stem cells undergoing differentiation should differ in many aspects, such as morphology and possibly also behaviour; however, such a correlation has not yet been exploited for fate prediction of MSCs. Primary human MSCs from bone marrow were expanded and pelleted to form high-density cultures and were then randomly divided into four groups to differentiate into adipogenic, osteogenic chondrogenic and myogenic progenitor cells. The cells were expanded as heterogeneous and tracked with time-lapse microscopy to record cell shape, using phase-contrast microscopy. The cells were segmented using a custom-made image-processing pipeline. Seven morphological features were extracted for each of the segmented cells. Statistical analysis was performed on the seven-dimensional feature vectors, using a tree-like classification method. Differentiation of cells was monitored with key marker genes and histology. Cells in differentiation media were expressing the key genes for each of the three pathways after 21 days, i.e. adipogenic, osteogenic and chondrogenic, which was also confirmed by histological staining. Time-lapse microscopy data were obtained and contained new evidence that two cell shape features, eccentricity and filopodia (= 'fingers') are highly informative to classify myogenic differentiation from all others. However, no robust classifiers could be identified for the other cell differentiation paths. The results suggest that non-invasive automated time-lapse microscopy could potentially be used to predict the stem cell fate of hMSCs for clinical application, based on morphology for earlier time-points. The classification is challenged by cell density, proliferation and possible unknown donor-specific factors, which affect the performance of morphology-based approaches. Copyright © 2012 John Wiley & Sons, Ltd.

Response to Innes and Mattsson: Neospora caninum emerges from the shadow of Toxoplasma gondii

It is tempting to extrapolate research findings regarding the intensively studied Toxoplasma gondii to Neospora caninum. This is based on morphological and ultrastructural studies, the molecular phylogeny of both parasites, their wide host ranges in nature, their ability to invade many different cell types in vitro and the occurrence of homologous proteins in both species. However, as Innes and Mattsson point out, T. gondii is the most successful parasite worldwide, whereas N. caninum has a more limited host range. Thus, some of the most challenging questions are: (i) what is T. gondii doing that N. caninum is not doing, or is doing differently, that renders the former so much more successful? And (ii) can some of these features be exploited for the development of interventional tools to limit infection and pathology caused by N. caninum?

Metabolites from intestinal microbes shape Treg

Intestinal bacterial metabolites are an important communication tool between the host immune system and the commensal microbiota to establish mutualism. In a recent paper published in Science, Wendy Garrett and her colleagues report an exciting role of the three most abundant microbial-derived short-chain fatty acids (SCFA), acetic acid, propionic acid and butyric acid, in colonic regulatory T cell (cTreg) homeostasis.

Stereological estimation of particle shape and orientation from volume tensors

In the present paper, we describe new robust methods of estimating cell shape and orientation in 3D from sections. The descriptors of 3D cell shape and orientation are based on volume tensors which are used to construct an ellipsoid, the Miles ellipsoid, approximating the average cell shape and orientation in 3D. The estimators of volume tensors are based on observations in several optical planes through sampled cells. This type of geometric sampling design is known as the optical rotator. The statistical behaviour of the estimator of the Miles ellipsoid is studied under a flexible model for 3D cell shape and orientation. In a simulation study, the lengths of the axes of the Miles ellipsoid can be estimated with CVs of about 2% if 100 cells are sampled. Finally, we illustrate the use of the developed methods in an example, involving neurons in the medial prefrontal cortex of rat.

(Table 1) Stratigraphic position, abundance, shape, color, and refractive indices of glass shards of selected samples from DSDP Leg 60 Holes vitric muds

Precisely determined refractive indices of glass shards from 32 ash-rich, volcaniclastic sediments, mostly turbidites interbedded with nonvolcanic sediments in the Mariana Trough, range from 1.480 to 1.585 (corresponding to SiO2 ca. 75 to 49%), with most in the range 1.500 to 1.540 (SiO2 ca. 70-62%) and a second, smaller mode between ca. 1.560 and 1.585 (57 to 49% SiO2). Shards are almost exclusively colorless from 1.480 to ca. 1.530, light brown with minor colorless and green tones between 1.530 and 1.560, and dominantly brown at higher refractive indices. Tubular pumice shards are more common at higher silica percentages and non- to poorly-vesicular cuniform shards at low SiO2 values, but there is no clear correlation between shape and composition of shards. About half of the samples have bimodal shard populations with silica differences ranging up to 20 percent; unimodal layers have a range of up to about 7 percent SiO2. Of 21 samples in which one type of shard dominates, seven have the main mode in the rhyolitic composition (>69% SiO2), eight in the intermediate range (56 to 69% SiO2), and five in mafic composition (SiO2 <53%). These unusually abundant mafic shards occur mainly in site survey piston cores, SP-IA and 4E, and in Holes 454, 456, 458, and 459B. These are the sites closest to the present arc. Hole 453, containing by far the most vitric tuff turbidites, shows a gradual increase in silica content of ash layers upward to the hole from Cores 36 to 19 (about 4.6 to 3.0 Ma). A drastic decrease in ash-rich beds in the younger (Pleistocene) part of this hole was noted by the shipboard party (see site chapter, Site 453) and was interpreted by them as indicating increasing distance from the arc volcanoes as the trough opened. The increase in silica in ashes from the early to the late Pliocene at Site 453 could be interpreted in the same way and might indicate that the trough started to open in early Pliocene time.