Isolation of neural stem and oligodendrocyte progenitor cells from the brain of live rats

Summary Postnatal brain neural stem and progenitor cells (NSPCs) cluster in anatomically inaccessible stem cell niches, such as the subependymal zone (SEZ). Here, we describe a method for the isolation of NSPCs from live animals, which we term “milking.” The intracerebroventricular injection of a release cocktail, containing neuraminidase, integrin-β1-blocking antibody, and fibroblast growth factor 2, induces the controlled flow of NSPCs in the cerebrospinal fluid, where they are collected via liquid biopsies. Isolated cells retain key in vivo self-renewal properties and their cell-type profile reflects the cell composition of their source area, while the function of the niche is sustained even 8 months post-milking. By changing the target area more caudally, we also isolate oligodendrocyte progenitor cells (OPCs) from the corpus callosum. This novel approach for sampling NSPCs and OPCs paves the way for performing longitudinal studies in experimental animals, for more in vivo relevant cell culture assays, and for future clinical neuro-regenerative applications

Being a Neural Stem Cell: A Matter of Character But Defined by the Microenvironment

The cells that build the nervous system, either this is a small network of ganglia or a complicated primate brain, are called neural stem and progenitor cells. Even though the very primitive and the very recent neural stem cells (NSCs) share common basic characteristics that are hard-wired within their character, such as the expression of transcription factors of the SoxB family, their capacity to give rise to extremely different neural tissues depends significantly on instructions from the microenvironment. In this chapter we explore the nature of the NSC microenvironment, looking through evolution, embryonic development, maturity and even disease. Experimental work undertaken over the last 20 years has revealed exciting insight into the NSC microcosmos. NSCs are very capable in producing their own extracellular matrix and in regulating their behaviour in an autocrine and paracrine manner. Nevertheless, accumulating evidence indicates an important role for the vasculature, especially within the NSC niches of the postnatal brain; while novel results reveal direct links between the metabolic state of the organism and the function of NSCs

A genetic mammalian proportional-integral feedback control circuit for robust and precise gene regulation

The processes that keep a cell alive are constantly challenged by unpredictable changes in its environment. Cells manage to counteract these changes by employing sophisticated regulatory strategies that maintain a steady internal milieu. Recently, the antithetic integral feedback motif has been demonstrated to be a minimal and universal biological regulatory strategy that can guarantee robust perfect adaptation for noisy gene regulatory networks in Escherichia coli. Here, we present a realization of the antithetic integral feedback motif in a synthetic gene circuit in mammalian cells. We show that the motif robustly maintains the expression of a synthetic transcription factor at tunable levels even when it is perturbed by increased degradation or its interaction network structure is perturbed by a negative feedback loop with an RNA-binding protein. We further demonstrate an improved regulatory strategy by augmenting the antithetic integral motif with additional negative feedback to realize antithetic proportional-integral control. We show that this motif produces robust perfect adaptation while also reducing the variance of the regulated synthetic transcription factor. We demonstrate that the integral and proportional-integral feedback motifs can mitigate the impact of gene expression burden, and we computationally explore their use in cell therapy. We believe that the engineering of precise and robust perfect adaptation will enable substantial advances in industrial biotechnology and cell-based therapeutics.ISSN:0027-8424ISSN:1091-649

ParlAmI: A Multimodal Approach for Programming Intelligent Environments

The proliferation of Internet of Things (IoT) devices and services and their integration in intelligent environments creates the need for a simple yet effective way of controlling and communicating with them. Towards such a direction, this work presents ParlAmI, a conversational framework featuring a multimodal chatbot that permits users to create simple “if-then” rules to define the behavior of an intelligent environment. ParlAmI delivers a disembodied conversational agent in the form of a messaging application named MAI, and an embodied conversational agent named nAoMI employing the programmable humanoid robot NAO. This paper describes the requirements and architecture of ParlAmI, the infrastructure of the “Intelligent Home” in which ParlAmI is deployed, the characteristics and functionality of both MAI and nAoMI, and finally presents the findings of a user experience evaluation that was conducted with the participation of sixteen users

Isolation of neural stem and oligodendrocyte progenitor cells from the brain of live rats.

Postnatal brain neural stem and progenitor cells (NSPCs) cluster in anatomically inaccessible stem cell niches, such as the subependymal zone (SEZ). Here, we describe a method for the isolation of NSPCs from live animals, which we term "milking." The intracerebroventricular injection of a release cocktail, containing neuraminidase, integrin-β1-blocking antibody, and fibroblast growth factor 2, induces the controlled flow of NSPCs in the cerebrospinal fluid, where they are collected via liquid biopsies. Isolated cells retain key in vivo self-renewal properties and their cell-type profile reflects the cell composition of their source area, while the function of the niche is sustained even 8 months post-milking. By changing the target area more caudally, we also isolate oligodendrocyte progenitor cells (OPCs) from the corpus callosum. This novel approach for sampling NSPCs and OPCs paves the way for performing longitudinal studies in experimental animals, for more in vivo relevant cell culture assays, and for future clinical neuro-regenerative applications

Ambient Intelligence in the Living Room

The emergence of the Ambient Intelligence (AmI) paradigm and the proliferation of Internet of Things (IoT) devices and services unveiled new potentials for the domain of domestic living, where the line between “the computer” and the (intelligent) environment becomes altogether invisible. Particularly, the residents of a house can use the living room not only as a traditional social and individual space where many activities take place, but also as a smart ecosystem that (a) enhances leisure activities by providing a rich suite of entertainment applications, (b) implements a home control middleware, (c) acts as an intervention host that is able to display appropriate content when the users need help or support, (d) behaves as an intelligent agent that communicates with the users in a natural manner and assists them throughout their daily activities, (e) presents a notification hub that provides personalized alerts according to contextual information, and (f) becomes an intermediary communication center for the family. This paper (i) describes how the “Intelligent Living Room” realizes these newly emerged roles, (ii) presents the process that was followed in order to design the living room environment, (iii) introduces the hardware and software facilities that were developed in order to improve quality of life, and (iv) reports the findings of various evaluation experiments conducted to assess the overall User Experience (UX)