Studying therapeutic potentials of human mesenchymal stromal cells with Cas9/AAV-based genome editing
Abstract/Contents
- Abstract
- Human mesenchymal stromal cells (hMSCs) are a promising source for cell-based therapies that require short-term interventions in injured tissues without replacement of resident cells. Genetic engineering of hMSCs could elucidate their mechanisms of function, enhance their therapeutic efficacy, and install novel functions, yet existing methods for MSCs are limited in their site specificity and ease of design, which ultimately constraint their applicability to research and clinical uses. Here, we describe an optimized Cas9/AAV6-based genome editing tool platform for hMSCs. Our results show that the system is applicable to cells derived from the bone marrow, adipose tissue, and umbilical cord blood. We successfully integrated more than 3 kilobases of exogenous DNA sequence into the hMSC genome without altering ex vivo characteristics of the cells. As a proof of concept, we applied the technique to generate in vivo bioluminescent hMSCs for biodistribution studies following infusion and local injection, and produced therapeutic factor-secreting MSCs that can exert enhanced therapeutic efficacy for wound healing in diabetic (db/db) mice over non-edited hMSCs. The genome editing platform is versatile in its application and is compatible with existing tissue engineering approaches for wound treatment and beyond. This study demonstrates the potential for precise genetic engineering of hMSCs and establishes a platform for future development of mechanism of action studies and engineered MSC-based therapies.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2019; ©2019 |
| Publication date | 2019; 2019 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Srifa, Waracharee |
|---|---|
| Degree supervisor | Porteus, Matthew H |
| Thesis advisor | Porteus, Matthew H |
| Thesis advisor | Gurtner, Geoffrey |
| Thesis advisor | Longaker, Michael T |
| Degree committee member | Gurtner, Geoffrey |
| Degree committee member | Longaker, Michael T |
| Associated with | Stanford University, Program in Stem Cell Biology and Regenerative Medicine. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Waracharee Srifa. |
|---|---|
| Note | Submitted to the Program in Stem Cell Biology and Regenerative Medicine. |
| Thesis | Thesis Ph.D. Stanford University 2019. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2019 by Waracharee Srifa
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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