Morphological Comparison of Stem Cells Using Two- Dimensional Culture and Spheroid Culture

Abstract: This chapter takes the reader on an imaginary scuba diving tour of the watery world of the cell and its surroundings, pointing out features such as the cytoskeleton (that forms the equivalent of the bones and muscles of our cells), the cell membrane (the outer skin of the cell), and the cell membrane’s embedded proteins that provide selective access to the interior of the cell—organelles (elfin versions our own organs). The chapter stresses the tumultuous action that occurs non-stop within the cells as proteins are assembled for use within and outside the cells. The chapter discusses stem cells, including the discovery of induced pluripotent stem cells. The chapter relates how cells differentiate to become dissimilar cell types, stresses the importance of three-dimensional study of cells (rather than two-dimensional study), and explains the different ways in which cells talk to each other.

Download Full-text

Modeling Herpes Simplex Virus 1 Infections in Human Central Nervous System Neuronal Cells Using Two- and Three-Dimensional Cultures Derived from Induced Pluripotent Stem Cells

Journal of Virology ◽

10.1128/jvi.00111-19 ◽

2019 ◽

Vol 93 (9) ◽

Cited By ~ 11

Author(s):

Leonardo D’Aiuto ◽

David C. Bloom ◽

Jennifer N. Naciri ◽

Adam Smith ◽

Terri G. Edwards ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Three Dimensional ◽

Two Dimensional ◽

Herpes Simplex Virus 1 ◽

Cns Neurons ◽

Induced Pluripotent ◽

Simplex Virus ◽

Hsv 1

ABSTRACTHerpes simplex virus 1 (HSV-1) establishes latency in both peripheral nerve ganglia and the central nervous system (CNS). The outcomes of acute and latent infections in these different anatomic sites appear to be distinct. It is becoming clear that many of the existing culture models using animal primary neurons to investigate HSV-1 infection of the CNS are limited and not ideal, and most do not recapitulate features of CNS neurons. Human induced pluripotent stem cells (hiPSCs) and neurons derived from them are documented as tools to study aspects of neuropathogenesis, but few have focused on modeling infections of the CNS. Here, we characterize functional two-dimensional (2D) CNS-like neuron cultures and three-dimensional (3D) brain organoids made from hiPSCs to model HSV-1–human–CNS interactions. Our results show that (i) hiPSC-derived CNS neurons are permissive for HSV-1 infection; (ii) a quiescent state exhibiting key landmarks of HSV-1 latency described in animal models can be established in hiPSC-derived CNS neurons; (iii) the complex laminar structure of the organoids can be efficiently infected with HSV, with virus being transported from the periphery to the central layers of the organoid; and (iv) the organoids support reactivation of HSV-1, albeit less efficiently than 2D cultures. Collectively, our results indicate that hiPSC-derived neuronal platforms, especially 3D organoids, offer an extraordinary opportunity for modeling the interaction of HSV-1 with the complex cellular and architectural structure of the human CNS.IMPORTANCEThis study employed human induced pluripotent stem cells (hiPSCs) to model acute and latent HSV-1 infections in two-dimensional (2D) and three-dimensional (3D) CNS neuronal cultures. We successfully established acute HSV-1 infections and infections showing features of latency. HSV-1 infection of the 3D organoids was able to spread from the outer surface of the organoid and was transported to the interior lamina, providing a model to study HSV-1 trafficking through complex neuronal tissue structures. HSV-1 could be reactivated in both culture systems; though, in contrast to 2D cultures, it appeared to be more difficult to reactivate HSV-1 in 3D cultures, potentially paralleling the low efficiency of HSV-1 reactivation in the CNS of animal models. The reactivation events were accompanied by dramatic neuronal morphological changes and cell-cell fusion. Together, our results provide substantive evidence of the suitability of hiPSC-based neuronal platforms to model HSV-1–CNS interactions in a human context.

Download Full-text

M177. THE PROTEOME OF OLIGODENDROCYTES DIFFERENTIATED FROM NEURAL STEM CELL DERIVED FROM SCHIZOPHRENIA PATIENTS

Schizophrenia Bulletin ◽

10.1093/schbul/sbaa030.489 ◽

2020 ◽

Vol 46 (Supplement_1) ◽

pp. S204-S204

Author(s):

Caroline Teles Rodrigues ◽

Daniel Martins-De-Souza

Keyword(s):

Quality Of Life ◽

Signal Transduction ◽

Stem Cells ◽

Two Dimensional ◽

Rna Transport ◽

Differentially Expressed Proteins ◽

Functional Studies ◽

Canonical Pathways ◽

Biochemical Mechanisms

Abstract Background Oligodendrocytes constitute the majority of the cells in white matter and alterations in this region have been reported in patients with schizophrenia. The myelin sheath is produced exclusively by mature oligodendrocytes. Thus, a dysfunction during the maturation of these cells could lead to a change in normal myelination processes, causing the hypomyelination observed in patients with schizophrenia. In this manner, functional studies are needed for a better connection of these different aspects of the disease in order to understand the pathophysiology of schizophrenia in an integrated manner. Thus, our aim was to evaluate the differences between the proteome of oligodendrocytes derived from neural stem cells (NSCs) from patients with schizophrenia and controls. Methods The cells were differentiated using the protocol described by Yan, Shin, Jha and collaborators (Yan, Shin, Jha, et al., 2013). After 14 days, the proteins were extracted and proteomic analyses were performed in a two-dimensional microUPLC coupled to nano ESI-Q-IM-TOF mass spectrometer. Progenesis® QI software was used in order to identify and quantify the proteins. Results On average, 2046 proteins were identified and 444 had alterations in the expression levels. These differentially expressed proteins were related most with metabolism, RNA transport, spliceosome machinery, vesicular transport, and signal transduction. Discussion The proteins and canonical pathways found here may contribute to understanding the biochemical mechanisms involved in the disorder, which may provide new targets for the development of more effective treatments, improving the schizophrenic patient’s quality of life.

Download Full-text