Human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) hold significant potential for use in cellular therapies to treat a variety of currently incurable diseases, such as age-related macular degeneration (AMD). AMD is the leading cause of blindness in individuals over the age of 60, and this loss of vision results from the death and dysfunction of a specific group of cells, the retinal pigmented epithelium (RPE). hESCs and iPSCs can differentiate into RPE cells, although the standard differentiation process is time-consuming and inefficient. Additionally, standard hESC and iPSC culture systems used to maintain undifferentiated cells, and for differentiation to RPE cells, contain animal products, feeder fibroblast cells, and other components not amendable for clinical applications. In efforts to translate human pluripotent stem cell research to the clinic, feeder-free, defined, and xeno-free culture systems are being rapidly explored. These efforts have highlighted the importance of understanding how these cells interact with their environment, particularly with the extracellular matrix (ECM), during both undifferentiated growth and differentiation to desired cell types. This dissertation presents the novel use of ECM proteins and ECM-derived substrates for hESC and iPSC culture, for both maintaining these cells in an undifferentiated state and for improving clinical derivation of RPE cells. Undifferentiated hESCs and iPSCs can be maintained on different ECM proteins, for which they express specific integrin heterodimers necessary for interaction between the cell and the ECM. By mimicking the ECM environment surrounding RPE in vivo, through analysis of integrin and ECM expression, ECM proteins important for more defined RPE differentiation and maintenance can be identified and utilized in vitro. Defined and xeno-free culture systems, recently made commercially available for the undifferentiated culture of hESCs and iPSCs, are also supportive of RPE derivation from human pluripotent stem cells. Taken together, these findings emphasize the importance of the ECM in hESC and iPSC culture, and provide a basis for RPE derivation that may be suitable for the clinical translation of cellular treatments for retinal diseases, such as AMD.
|Advisor:||Clegg, Dennis O.|
|Commitee:||Foltz, Kathy, Hawker, Craig, Johnson, Lincoln|
|School:||University of California, Santa Barbara|
|Department:||Molecular, Cellular & Developmental Biology|
|School Location:||United States -- California|
|Source:||DAI-B 73/03, Dissertation Abstracts International|
|Keywords:||Age-related macular degeneration, Extracellular matrix, Integrins, Pluripotency, Retinal pigmented epithelium, Stem cells, Vitronectin|
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