Related Experiment Video
Updated: May 21, 2026
11:26
Isolation and Immortalization of Patient-derived Cell Lines from Muscle Biopsy for Disease Modeling
Published on: January 18, 2015
Inducible immortality in hTERT-human mesenchymal stem cells
Samantha L Piper1, Miqi Wang, Akira Yamamoto
1Department of Orthopaedic Surgery, University of California San Francisco, 500 Parnassus Avenue, San Francisco, California 94143, USA.
Summary
Human mesenchymal stem cells (hMSCs) can be engineered for extended proliferation using inducible telomerase. This controlled approach overcomes senescence, enabling longer cell culture for regenerative medicine applications.
Area of Science:
- Stem Cell Biology
- Regenerative Medicine
- Molecular Biology
Background:
- Human mesenchymal stem cells (hMSCs) are valuable for tissue engineering due to multipotency.
- Replicative senescence limits in vitro expansion and differentiation of hMSCs.
- Constitutive telomerase expression enhances proliferation but risks uncontrolled growth and tumor formation.
Purpose of the Study:
- To develop a method for controlled, long-term expansion of hMSCs.
- To investigate the role of inducible telomerase in overcoming hMSC senescence.
- To assess the safety and efficacy of telomerase-engineered hMSCs for therapeutic applications.
Main Methods:
- Utilized a Tet-On inducible gene expression system to control telomerase (hTERT) expression in hMSCs.
- Assessed proliferation capacity, telomere length, and multipotency of inducible hTERT hMSCs (i-hTERT hMSCs).
- Employed a selective telomere elongation inhibitor to confirm the mechanism of immortalization.
Main Results:
- i-hTERT hMSCs exhibited significantly longer proliferation than wild-type hMSCs upon doxycycline induction.
- Telomerase expression could be reversibly controlled, halting and resuming proliferation as needed.
- Extended-lifespan i-hTERT hMSCs retained in vitro multipotency and showed telomere length-dependent immortalization.
Conclusions:
- Inducible telomerase expression provides a safe and effective strategy for extending hMSC lifespan.
- i-hTERT hMSCs maintain essential stem cell properties, offering a promising tool for regenerative medicine.
- Controlled immortalization of hMSCs is dependent on telomere maintenance, mitigating risks of uncontrolled growth.
Related Concept Videos
01:19Multipotency of Hematopoietic Stem Cells
The hematopoietic stem cells or HSCs are multipotent, meaning they can differentiate and give rise to all blood and immune cells. HSCs are maintained in the quiescent stage until an external stimulus initiates their differentiation. The multipotent HSCs exist as two heterogeneous populations, long-term repopulating cells (LTRC) and short-term repopulating cells (STRC). The two HSC populations have different surface markers or receptors and are classified based on quiescence and long-term...
02:15Replicative Cell Senescence
Replicative cell senescence is a property of cells that allows them to divide a finite number of times throughout the organism's lifespan while preventing excessive proliferation. Replicative senescence is associated with the gradual loss of the telomere — short, repetitive DNA sequences found at the end of the chromosomes. Telomeres are bound by a group of proteins to form a protective cap on the ends of chromosomes. Embryonic stem cells express telomerase — an enzyme that adds the telomeric...
01:06Induced Pluripotent Stem Cells
Stem cells are undifferentiated cells that divide and produce different cell types. Ordinarily, cells that have differentiated into a specific cell type are terminally differentiated; however, scientists have found a way to reprogram these mature cells so that they dedifferentiate and return to an unspecialized, proliferative state. These cells are pluripotent like embryonic stem cells—able to produce all cell types—and are called induced pluripotent stem cells (iPSCs).
Somatic cells are...
Somatic cells are...
01:13Induced Pluripotent Stem Cells
Stem cells are undifferentiated cells that divide and produce different types of cells. Ordinarily, cells that have differentiated into a specific cell type are post-mitotic—that is, they no longer divide. However, scientists have found a way to reprogram these mature cells so that they “de-differentiate” and return to an unspecialized, proliferative state. These cells are also pluripotent like embryonic stem cells—able to produce all cell types—and are therefore called induced pluripotent stem...
01:19Mesenchymal Stem Cells
Mesenchymal stem cells (MSCs) are adult stem cells that can differentiate into most connective tissue cell types, except for hematopoietic cells, depending upon the source of MSCs. For example, bone-marrow-derived MSCs (BM-MSCs) can differentiate into osteocytes, hepatocytes, and pancreatic and neuronal cells. MSCs can be isolated from various sources such as bone marrow, placenta, adipose tissue, teeth, and Wharton’s jelly, a gelatinous substance in the umbilical cord. The ease of their access...
01:14Maintenance of the ES Cell State
The cells of the blastocyst inner cell mass only remain pluripotent for a short time. This state of pluripotency and self-renewal can be maintained in embryonic stem (ES) cell culture by adding specific chemicals or growth factors to ensure the cells can continue dividing and later differentiate into different cell types. In some cases, the cells are grown on a feeder layer of differentiated cells, which provides the growth factors and extracellular matrix components necessary for stem cell...