رسالة جامعية
Enabling the Next Generation of Human Induced Pluripotent Stem Cell Derived Hematopoietic Stem Cell-Based Therapies
| العنوان: | Enabling the Next Generation of Human Induced Pluripotent Stem Cell Derived Hematopoietic Stem Cell-Based Therapies |
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| المؤلفون: | Wong, Casey |
| مرشدي الرسالة: | Jezierski, Anna, Wang, Lisheng |
| بيانات النشر: | Université d'Ottawa / University of Ottawa, 2023. |
| سنة النشر: | 2023 |
| المجموعة: | Université d'Ottawa |
| مصطلحات موضوعية: | Stem cell agonist cocktail, Small molecule, Human induced pluripotent stem cell, Hematopoietic stem progenitor cell, Multipotent progenitor cell, Expansion, StemRegenin 1, UM171, Valproic acid, AA2P |
| الوصف: | Human induced pluripotent stem cells (iPSCs) represent a scalable cell source for the generation of hematopoietic progenitor cells (iHPCs); however, a lack of efficient iHPC expansion in vitro currently limits translational applications. To address this translational bottleneck, we assessed a panel of stem cell agonist cocktails (SCACs), originally developed to enhance cord-blood derived HSPC (CB-HSPC) expansion, on iHPC expansion. Three SCACs and GAS6 (X2A, X2A+GAS6, SM6, or SMA) were supplemented during iHPC differentiation and subsequent expansion using the STEMdiff™ Hematopoietic Kit. This monolayer differentiation strategy yielded a population of CD34⁺CD43⁺ and CD45⁺CD34⁺ iHPC. SCAC supplementation during iHPC differentiation yielded up to 2.5-fold higher frequency of CD34⁺CD43⁺ hematopoietic progenitors and up to 2.9-fold higher frequency of CD45⁺CD34⁺CD45RA⁻CD90⁺ HSC-like cells compared to non-treated controls. Subsequent SCAC supplementation during 2 weeks of expansion culture also significantly increased iHPC expansion (X2A+GAS6: 3.8-fold, X2A: 3.5-fold, SM6: 2.8-fold, SMA: 2.0-fold). The expanded iHPCs retained high levels of CD34⁺CD43⁺ expression but we observed an increase in the expansion of HSC-like cell fraction. The collective expansion observed with the SCACs was 1.5- to 2.8-fold higher than UM171 treatment alone. Furthermore, all SCAC-supplemented iHPCs retained multilineage potency, producing erythroid and granulocyte-macrophage progenitors in CFU assays. However, prolonged expansion, beyond 7 days, reduced multilineage potential, indicating a limited expansion window. Although optimal timing and composition of SCAC supplementation remains to be refined, these results highlight that exploiting the additive and synergistic effects of multiple small molecules represents a promising approach for enhancing iHPC expansion yields and biomanufacturing. |
| Original Identifier: | oai:ruor.uottawa.ca:10393/45319 |
| نوع الوثيقة: | Thesis |
| وصف الملف: | application/pdf |
| اللغة: | English |
| DOI: | 10.20381/ruor-29525 |
| الإتاحة: | http://hdl.handle.net/10393/45319Test |
| حقوق: | Attribution-ShareAlike 4.0 International URL: http://creativecommons.org/licenses/by-sa/4.0Test/ |
| رقم الانضمام: | edsndl.uottawa.ca.oai.ruor.uottawa.ca.10393.45319 |
| قاعدة البيانات: | Networked Digital Library of Theses & Dissertations |
| DOI: | 10.20381/ruor-29525 |
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