Biophysical Microenvironment Research Articles

Stem cell maintenance in a different niche

To overcome the difficulty of controlling stem cell fate and function in applications to regenerative medicine, a number of alternative approaches have been made. Recent reports demonstrate that a non-cellular niche modulating the biophysical microenvironment with chemical factors can support stem cell self-renewal. In our previous studies, early establishment was executed to optimize biophysical factors and it was subsequently found that the microgeometry of the extracellular matrix made huge differences in stem cell behavior and phenotype. We review here a three-dimensional, non-cellular niche designed to support stem cell self-renewal. The characteristics of stem cells under the designed system are further discussed.

Preparation of Hydrogel Substrates with Tunable Mechanical Properties

The modulus of elasticity of the extracellular matrix (ECM), often referred to in a biological context as "stiffness," naturally varies within the body, e.g., hard bones and soft tissue. Moreover, it has been found to have a profound effect on the behavior of anchorage-dependent cells. The fabrication of matrix substrates with a defined modulus of elasticity can be a useful technique to study the interactions of cells with their biophysical microenvironment. Matrix substrates composed of polyacrylamide hydrogels have an easily quantifiable elasticity that can be changed by adjusting the relative concentrations of its monomer, acrylamide, and cross-linker, bis-acrylamide. In this unit, we detail a protocol for the fabrication of statically compliant and radial-gradient polyacrylamide hydrogels, as well as the functionalization of these hydrogels with ECM proteins for cell culture. Included as well are suggestions to optimize this protocol to the choice of cell type or stiffness with a table of relative bis-acrylamide and acrylamide concentrations and expected elasticity after polymerization.

Functional Adaptation: The Key to Plasticity of Cardiovascular "Stem" Cells?

There is increasing evidence that cells of disparate phenotypes displaying various degrees of proliferative capacity engraft and function heterotopically in adult organisms. Efforts were made to reconcile these findings with the embryologic notions of pluripotent stem or progenitor cell, although the nature of the 'stemness' remained elusive. This topic is particularly important for the cardiovascular system, in which cytotrophoblasts, certain tumor cells, monocytes/macrophages, peritoneal mesothelial cells, and others acquire endothelial properties and/or perform endothelial functions. Here we suggest that this pluripotency reflects a fundamental characteristic of cellular diversity, which is manifested as the adaptive response to a functional pressure exerted by the cell's biochemical and biophysical microenvironments that would drive their differentiation. In this model, differentiation is a dynamic, reversible, and open-ended process where the cells would maintain the flexibility to respond to changing environmental clues with a fine tuning of their structure, a property that was previously called cellular plasticity. Pluripotent adult stem cells that display this property in culture, and, perhaps upon in vivo administration, were described. Therefore, we also suggest that differentiation of stem cells is a form of cellular plasticity within the larger context of functional adaptation, whereas their stemness remains associated with self-renewal.

Specific Docking of Apolipoprotein A-I at the Cell Surface Requires a Functional ABCA1 Transporter

The identification of defects in ABCA1 as the molecular basis of Tangier disease has highlighted its crucial role in the loading with phospholipids and cholesterol of nascent apolipoprotein particles. Indeed the expression of ABCA1 affects apolipoprotein A-I (apoA-I)-mediated removal of lipids from cell membranes, and the possible role of ABCA1 as an apoA-I surface receptor has been recently suggested. In the present study, we have investigated the role of the ABCA1 transporter as an apoA-I receptor with the analysis of a panel of transfectants expressing functional or mutant forms of ABCA1. We provide experimental evidence that the forced expression of a functional ABCA1 transporter confers surface competence for apoA-I binding. This, however, appears to be dependent on ABCA1 function. Structurally intact but ATPase-deficient forms of the transporter fail to elicit a specific cell association of the ligand. In addition the diffusion parameters of membrane-associated apoA-I indicate an interaction with membrane lipids rather than proteins. These results do not support a direct molecular interaction between ABCA1 and apoA-I, but rather suggest that the ABCA1-induced modification of the lipid distribution in the membrane, evidenced by the phosphatidylserine exofacial flopping, generates a biophysical microenvironment required for the docking of apoA-I at the cell surface.

Magnetic Resonance Imaging of the Common Marmoset Head

This study evaluated the changes in the intrinsic magnetic resonance (MR) relaxation parameter values (T1; T2, proton density, magnetisation transfer and apparent diffusion coefficient) of the marmoset head, imaged before and after death. Knowing the absolute values of the MR parameters makes it possible to choose an imaging protocol for optimal structural differentiation. The changes between the ante-mortem and post-mortem MR parameters provide an insight into the changing biophysical microenvironment of the post-mortem brain, and allow some of the changes that occur in pathological conditions to be predicted. Diffusion-weighted MR imaging (MRI) was used to map quantitative apparent diffusion coefficient values, and to investigate diffusional anisotropy along the fibre tracts in pre-mortem and post-mortem brain tissue. A three-dimensional data set of the entire marmoset brain demonstrates the ability of three-dimensional MRI to differentiate internal brain structures. MRI is a non-invasive technique which, in principle, permits the same animal to be re-imaged serially and has the potential to probe in vivo brain structural and biophysical changes over an extended period of time. Serial imaging, where each animal acts as its own control, reduces the number of animals required to detect a significant change by minimising the effects of inter-subject variance. MRI therefore provides important scientific and ethical benefits.