| Chapter title |
Induction and Detection of Autophagy in Aged Hematopoietic Stem Cells by Exposing Them to Microvesicles Secreted by HSC-Supportive Mesenchymal Stromal Cells
|
|---|---|
| Chapter number | 166 |
| Book title |
Autophagy in Differentiation and Tissue Maintenance
|
| Published in |
Methods in molecular biology, June 2018
|
| DOI | 10.1007/7651_2018_166 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-8747-4, 978-1-4939-8748-1
|
| Authors |
Rohan S. Kulkarni, Manmohan S. Bajaj, Vaijayanti P. Kale, Kulkarni, Rohan S., Bajaj, Manmohan S., Kale, Vaijayanti P. |
| Abstract |
Autophagy is an important cellular process for maintenance of quality and functionality of cells. This happens through repair and renewal of cellular components like proteins and mitochondria. Reduction in autophagy process in aged hematopoietic stem cells (HSCs) leads to their compromised stemness and self-renewal capacity, and consequently, their applicability in various regenerative therapies also reduces. HSC functions are regulated by their microenvironment, known as "HSC niche," which comprises of mesenchymal stromal cells (MSCs), osteoblasts, endothelial cells, etc. In this niche, the MSCs are known to closely interact with the HSCs, and therefore, they can directly influence the stem cell fate. In our earlier studies, we have demonstrated that young MSCs or aged MSCs rejuvenated by treating them with LY294002, a PI3K inhibitor (rescued aged MSCs), rejuvenate aged HSCs via intercellular transfer of microvesicles (MVs) harboring autophagy-inducing mRNAs.Here, we describe the protocol for induction of autophagy in aged HSCs by incubating them with microvesicles (MVs) collected from young MSCs or rescued aged MSCs. We also describe the protocols for determination of autophagy levels in these HSCs. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 2 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 50% |
| Science communicators (journalists, bloggers, editors) | 1 | 50% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 19 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 3 | 16% |
| Student > Master | 3 | 16% |
| Researcher | 2 | 11% |
| Student > Bachelor | 1 | 5% |
| Other | 1 | 5% |
| Other | 1 | 5% |
| Unknown | 8 | 42% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 8 | 42% |
| Nursing and Health Professions | 1 | 5% |
| Agricultural and Biological Sciences | 1 | 5% |
| Medicine and Dentistry | 1 | 5% |
| Unknown | 8 | 42% |