For the purpose of developing an effective therapeutic strategy for HSCR, the enteric neural crest stem cells were investigated firstly which were isolated from the E14.5 mouse embryonic gut, cultured as neurospheres and characterized by multiple immunofluorescence and reverse transcription-PCR, population doubling time, frequency of forming secondary neurospheres and limited dilution assay. In the differentiation culture medium, several types of cells were induced to form from the neurospheres derived from single cells. Hence the putative enteric neural crest stern cells, which were isolated from the embryonic mouse gut tube and cultured as neurospheres for many passages ex vivo with the demonstrated capacity of proliferation, self-renewal and differentiation, showed properties of stem cells. / Hirschsprung's disease (HSCR) is caused by the absence of the enteric neural crest-derived neurons at the distal region of the gut. Cell-based therapy using stem cells or progenitors gives the potential to supplement these missing enteric neurons in the gut. Enteric neural crest stem cells isolated from the human or rodent gut can give rise to neurons and glia after they are transplanted into the recipient guts of the mouse or rat. However, numbers of issues are unresolved about the basic biology of the enteric nervous system, the characteristics of the stem cells isolated from the enteric nervous system and the biological significance of these cells in prenatal and postnatal periods. In this study, the characteristics and therapeutic potential on HSCR of the enteric neural crest stem cells were explored. / In addition to the above, a recombination organotypic gut culture ex vivo showed that the colonization of enteric neural crest-derived cells in the recipient gut was influenced not only by the genotypes of enteric neural crest-derived cells themselves but also the microenvironment of the gut through which enteric neural crest-derived cells migrated. For instance, the developmental stage of the recipient gut and also the presence of endogenous enteric neural crest-derived cells along the migratory pathway of neural crest-derived cells both affected the extent of the migration and colonization of exogenous enteric neural crest-derived cells and stem cells. The gradual maturation and differentiation of the neighboring structures, such as the smooth muscle layer, during the time period of the enteric neural crest cells migration, might also suggest that these neighboring tissues may have a role in regulating the neural crest-derived cells migration. / In conclusion, enteric neural crest stem cells isolated from the embryonic mouse gut tube showed properties of stem cells, and had the potential to compensate missing enteric neural crest-derived cells both ex vivo and in vivo. However, the colonization of enteric neural crest-derived cells in the developing gut was affected cell-autonomously and also by the microenvironment of the gut and the presence of existing enteric neural crest-derived cells. / Their potential applications in the transplantation experiments were shown by transplantation of the neurospheres isolated to the gut tube maintained in an organotypic culture or to the descending colon of neonates at postnatal day 7. The development of the enteric neural crest stern cells from the neurospheres was found to be compatible to endogenous enteric neural crest-derived cells in the recipient gut as evidenced by the formation of interconnected cellular networks of donor stem cells and endogenous neural crest-derived cells. The enteric neural crest stem cells also possess the potential to compensate the loss of enteric neural crest-derived cells ex vivo and in vivo in recipient prenatal and postnatal guts. / Bao, Lihua. / Adviser: Wood Yee Chan. / Source: Dissertation Abstracts International, Volume: 73-01, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 208-228). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_344665 |
| Date | January 2010 |
| Contributors | Bao, Lihua., Chinese University of Hong Kong Graduate School. Division of Anatomy. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, theses |
| Format | electronic resource, microform, microfiche, 1 online resource (x, 228 leaves : ill.) |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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