Cell-cell and cell-medium interactions in the growth of mouse embryonic stem cells

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Physics, 2010. === This electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections. === Cataloged from student-submitted PDF version of thesis. === Inclu...

Full description

Bibliographic Details
Main Author: Mittal, Nikhil, 1979-
Other Authors: Joel Voldman and Alexander van Oudenaarden.
Format: Others
Language:English
Published: Massachusetts Institute of Technology 2011
Subjects:
Online Access:http://hdl.handle.net/1721.1/62602
id ndltd-MIT-oai-dspace.mit.edu-1721.1-62602
record_format oai_dc
spelling ndltd-MIT-oai-dspace.mit.edu-1721.1-626022019-05-02T15:57:00Z Cell-cell and cell-medium interactions in the growth of mouse embryonic stem cells Cell-cell and cell-medium interactions in the growth of mouse ESCs Mittal, Nikhil, 1979- Joel Voldman and Alexander van Oudenaarden. Massachusetts Institute of Technology. Dept. of Physics. Massachusetts Institute of Technology. Dept. of Physics. Physics. Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Physics, 2010. This electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections. Cataloged from student-submitted PDF version of thesis. Includes bibliographical references (p. 100-108). Embryonic stem cells serve as powerful models for the study of development and disease and hold enormous potential for future therapeutics. Due to the potential for embryonic stem cells (ESCs) to provide a variety of tissues for use in regenerative medicine, there has been great interest in the identification of factors that govern the differentiation of ESCs into specific lineages. Much of this research builds on previous studies of the role of intercellular signaling in the specification of various cell types in the developing embryo. However, relatively little work has been done on understanding the role of cell-cell communication in the self-renewal of ESCs. In the first part of this thesis I describe the development and testing of new devices for studying intercellular signaling - the nDEP microwell array and the Bio Flip Chip (BFC). We used the BFC to show that cell-cell interaction improves the colony-forming efficiency and the self-renewal of mouse ESCs. Further, we demonstrate that the interaction is at least partly diffusible. In the next part of the thesis I describe our use of more traditional assays to validate the results obtained using the BFC and to further explore the role of diffusible signaling in the survival of mouse ESCs. We demonstrate the existence of an optimal density for 2-day culture of mouse ESCs. Further, we demonstrate that the increase in growth with plating density (103-104 cells/cm2) is at least partly due to the existence of one or more survival-enhancing autocrine factor(s) in mouse ESC cultures, and that one of these factors is Cyclophilin A. Finally, we demonstrate that changes in the low molecular weight composition of the medium are likely responsible for the decrease in growth at high plating densities (>104 cells/cm2). We use a numerical model to show that competition between the positive effect (on growth) of autocrine survival factors and the negative effect of nutrient depletion can account for the observed optimal growth density. Our study provides new insight into the processes underlying, and optimization of, growth in cell types that lack contact inhibition such as cancer cells and stem cells. by Nikhil V. Mittal. Ph.D. 2011-05-09T13:59:15Z 2011-05-09T13:59:15Z 2010 2010 Thesis http://hdl.handle.net/1721.1/62602 713650794 eng M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission. http://dspace.mit.edu/handle/1721.1/7582 130 p. application/pdf Massachusetts Institute of Technology
collection NDLTD
language English
format Others
sources NDLTD
topic Physics.
spellingShingle Physics.
Mittal, Nikhil, 1979-
Cell-cell and cell-medium interactions in the growth of mouse embryonic stem cells
description Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Physics, 2010. === This electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections. === Cataloged from student-submitted PDF version of thesis. === Includes bibliographical references (p. 100-108). === Embryonic stem cells serve as powerful models for the study of development and disease and hold enormous potential for future therapeutics. Due to the potential for embryonic stem cells (ESCs) to provide a variety of tissues for use in regenerative medicine, there has been great interest in the identification of factors that govern the differentiation of ESCs into specific lineages. Much of this research builds on previous studies of the role of intercellular signaling in the specification of various cell types in the developing embryo. However, relatively little work has been done on understanding the role of cell-cell communication in the self-renewal of ESCs. In the first part of this thesis I describe the development and testing of new devices for studying intercellular signaling - the nDEP microwell array and the Bio Flip Chip (BFC). We used the BFC to show that cell-cell interaction improves the colony-forming efficiency and the self-renewal of mouse ESCs. Further, we demonstrate that the interaction is at least partly diffusible. In the next part of the thesis I describe our use of more traditional assays to validate the results obtained using the BFC and to further explore the role of diffusible signaling in the survival of mouse ESCs. We demonstrate the existence of an optimal density for 2-day culture of mouse ESCs. Further, we demonstrate that the increase in growth with plating density (103-104 cells/cm2) is at least partly due to the existence of one or more survival-enhancing autocrine factor(s) in mouse ESC cultures, and that one of these factors is Cyclophilin A. Finally, we demonstrate that changes in the low molecular weight composition of the medium are likely responsible for the decrease in growth at high plating densities (>104 cells/cm2). We use a numerical model to show that competition between the positive effect (on growth) of autocrine survival factors and the negative effect of nutrient depletion can account for the observed optimal growth density. Our study provides new insight into the processes underlying, and optimization of, growth in cell types that lack contact inhibition such as cancer cells and stem cells. === by Nikhil V. Mittal. === Ph.D.
author2 Joel Voldman and Alexander van Oudenaarden.
author_facet Joel Voldman and Alexander van Oudenaarden.
Mittal, Nikhil, 1979-
author Mittal, Nikhil, 1979-
author_sort Mittal, Nikhil, 1979-
title Cell-cell and cell-medium interactions in the growth of mouse embryonic stem cells
title_short Cell-cell and cell-medium interactions in the growth of mouse embryonic stem cells
title_full Cell-cell and cell-medium interactions in the growth of mouse embryonic stem cells
title_fullStr Cell-cell and cell-medium interactions in the growth of mouse embryonic stem cells
title_full_unstemmed Cell-cell and cell-medium interactions in the growth of mouse embryonic stem cells
title_sort cell-cell and cell-medium interactions in the growth of mouse embryonic stem cells
publisher Massachusetts Institute of Technology
publishDate 2011
url http://hdl.handle.net/1721.1/62602
work_keys_str_mv AT mittalnikhil1979 cellcellandcellmediuminteractionsinthegrowthofmouseembryonicstemcells
AT mittalnikhil1979 cellcellandcellmediuminteractionsinthegrowthofmouseescs
_version_ 1719031956487798784