Generation and functional characterization of PSC-CX3CR1+ cells Open Access

Andrade, Mark (Spring 2019)

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Cardiomyocytes (CM) in the adult mammalian heart have limited proliferative capabilities which result in the inability of the heart to regenerate. The field of stem/progenitor cell therapies had turned to cardiac progenitor cells as an essential entity to overcome the boundaries of mammalian heart regeneration. Modern biology has been able to identify many embryonic and postnatal stem/progenitor populations demonstrating cardiogenic properties. However, to date only embryonic cardiac progenitor cells remain non-controversial about their roles in cardiogenesis. Furthermore, studies revolving around these populations have many limitations and display shortcomings in clinical trials when compare to their preclinical results. Hence, it is evident that further functional characterization, including further identification of cardiovascular progenitor cells, for embryonic progenitor cells needs to be established before clinical trials results live up to their expectations. A recent study has revealed that macrophage precursors positive to CX3CR1 arise in the yolk sack and migrate to fetal organs during embryonic development.1 Additionally, in our lab, a cell lineage study has demonstrated that these cx3cr1 cell lines contribute not only to organogenesis but also to the cardiovascular system. Hence, here we establish a system by which, CX3CR1+ cells are derived from mouse embryonic stem cells and can contribute to the cardiovascular system. In vitro, under cardiac promoting media, they demonstrated direct differentiation into both cardiomyocytes (CMs) and endothelial cells (ECs). Ex vivo, co-culturing these cells with a fetal mouse heart demonstrated their ability to migrate toward the ex vivo fetal mouse heart and incorporate themselves into the CM population. In vivo, using a myocardial infarction (MI) mouse model, injection of these cells with biomaterial recapitulated both the in vitro and ex vivo results. This study adds insights to our understanding of multipotent cell lines and add to the advancement of stem cell therapy for heart regeneration. 

Table of Contents

Chapter 1: Introduction                                                                        1

Chapter 2: Results                                                                                3-14

  CX3CR1+ cells were generated from mouse embryonic stem cells            3

Figure 1. CX3CR1 expression kinetics on differentiating mESCs 4

mESC-derived CX3CR1+ cells retain capacity to differentiate into cardiovascular cell lineages in vitro 4

Figure 2. In vitro cardiovascular differentiation of mESC-derived CX3CR1+ cells  5-7

mESC-derived CX3CR1+ cells contributed to the cardiovascular lineage populations present 7

in the fetal mouse heart ex vivo

Figure 3. mESC-derived CX3CR1+ cells migration to the fetal heart and cardiovascular 8-10

commitment ex vivo

mESC-derived CX3CR1+ cells contributed to the cardiovascular lineage populations in 10

the post-MI adult mouse heart in vivo 

Chapter 3: Discussion                                                                          15-18

Chapter 4: Methods                                                                              19-21

Chapter 5: References                                                                          22

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