<abstract xmlns="http://www.w3.org/1999/xhtml"> Background: Human bone marrow stem cell (hBMSC) is a promising candidate in regenerative medicine due to its plasticity and homing ability. However, in vitro survival and therapeutic efficacy of hBMSCs decline with donor age as a result of physiological cellular aging. Objectives: We transfected human telomerase reverse transcriptase (hTERT) into hBMSCs from patients more than 60 years old in the hope of extending cellular life span. Methods: In our study on 11 donors (aged 16-74 years), no telomerase activity was detected in hBMSCs from donors ≥60 years old. We then performed transient transfection on hBMSC from these elderly donors (n = 4) with hTERT gene using non-viral methods. Results: Transfection efficiency of 38.5% and 55.6% was achieved by lipofection and electroporation with viability at 83% and 44%, respectively. Telomerase was detected on day 5 for all post-transfected samples. Maximum passage number achieved after transfection was ≥13 (≈45 doublings) compared with 6 (≈15 doublings) before transfection. Conclusion: We conclude that transient expression of hTERT increased cellular life span of hBMSCs from elderly patients, while maintaining a normal cell morphology and growth rate.</abstract>

Background: Human bone marrow stem cell (hBMSC) is a promising candidate in regenerative medicine due to its plasticity and homing ability. However, in vitro survival and therapeutic efficacy of hBMSCs decline with donor age as a result of physiological cellular aging. Objectives: We transfected human telomerase reverse transcriptase (hTERT) into hBMSCs from patients more than 60 years old in the hope of extending cellular life span. Methods: In our study on 11 donors (aged 16-74 years), no telomerase activity was detected in hBMSCs from donors ≥60 years old. We then performed transient transfection on hBMSC from these elderly donors (n = 4) with hTERT gene using non-viral methods. Results: Transfection efficiency of 38.5% and 55.6% was achieved by lipofection and electroporation with viability at 83% and 44%, respectively. Telomerase was detected on day 5 for all post-transfected samples. Maximum passage number achieved after transfection was ≥13 (≈45 doublings) compared with 6 (≈15 doublings) before transfection. Conclusion: We conclude that transient expression of hTERT increased cellular life span of hBMSCs from elderly patients, while maintaining a normal cell morphology and growth rate.

Non-viral transient transfection of hTERT gene into hBMSCs from elderly patients delays cellular aging in vitro

Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Bandar Tun Razak, Cheras 56000, Kuala Lumpur,

Ear, Nose and Throat Consultant Clinic, Ampang Puteri Specialist Hospital, Selangor 68000, Kuala Lumpur 50300,

Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Bandar Tun Razak, Cheras 56000, Kuala Lumpur, Malaysia

Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur 50300,

Asian Biomedicine

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Background: Human bone marrow stem cell (hBMSC) is a promising candidate in regenerative medicine due to its plasticity and homing ability. However, in vitro...

Non-viral transient transfection of hTERT gene into hBMSCs from elderly patients delays cellular aging in vitro

Article Category: Original article

Published Online: Feb 04, 2017

Page range: 245 - 254

DOI: https://doi.org/10.5372/1905-7415.0602.062

Keywords
Cellular life span, human bone marrow stem cell, telomerase, transfection

© 2017

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Non-viral transient transfection of hTERT gene into hBMSCs from elderly patients delays cellular aging in vitro

Article Category: Original article

Published Online: Feb 04, 2017

Page range: 245 - 254

DOI: https://doi.org/10.5372/1905-7415.0602.062

KeywordsCellular life span, human bone marrow stem cell, telomerase, transfection

© 2017

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Keywords
Cellular life span, human bone marrow stem cell, telomerase, transfection