Int J Stem Cells.  2016 May;9(1):107-114. 10.15283/ijsc.2016.9.1.107.

L-carnitine Effectively Induces hTERT Gene Expression of Human Adipose Tissue-derived Mesenchymal Stem Cells Obtained from the Aged Subjects

• Farahzadi
• Mesbah-Namin SA
• Zarghami N
• Fathi E

• KMID: 2164167
• DOI: http://doi.org/10.15283/ijsc.2016.9.1.107

Abstract

BACKGROUND AND OBJECTIVES
Human mesenchymal stem cells (hMSCs) are attractive candidates for cell therapy and regenerative medicine due to their multipotency and ready availability, but their application can be complicated by the factors such as age of the donors and senescence-associated growth arrest during culture conditions. The latter most likely reflects the fact that aging of hMSCs is associated with a rise in intracellular reactive oxygen species, loss of telomerase activity, decrease in human telomerase reverse transcriptase (hTERT) expression and finally eroded telomere ends. Over-expression of telomerase in hMSCs leads to telomere elongation and may help to maintain replicative life-span of these cells. The aim of this study was to evaluate of the effect of L-carnitine (LC) as an antioxidant on the telomerase gene expression and telomere length in aged adipose tissue-derived hMSCs.
METHODS
For this purpose, cells were isolated from healthy aged volunteers and their viabilities were assessed by MTT assay. Quantitative gene expression of hTERT and absolute telomere length measurement were also performed by real- time PCR in the absence and presence of different doses of LC (0.1, 0.2 and 0.4 mM).
RESULTS
The results indicated that LC could significantly increase the hTERT gene expression and telomere length, especially in dose of 0.2 mM of LC and in 48 h treatment for the aged adipose tissue-derived hMSCs samples.
CONCLUSION
It seems that LC would be a good candidate to improve the lifespan of the aged adipose tissue-derived hMSCs due to over-expression of telomerase and lengthening of the telomeres.

Keyword

L-carnitine; hTERT gene expression; Human MSCs; Antioxidant

MeSH Terms

Aging
Carnitine*
Cell- and Tissue-Based Therapy
Cytochrome P-450 CYP1A1
Gene Expression*
Humans*
Mesenchymal Stromal Cells*
Polymerase Chain Reaction
Reactive Oxygen Species
Regenerative Medicine
Telomerase
Telomere
Tissue Donors
Volunteers
Carnitine
Cytochrome P-450 CYP1A1
Reactive Oxygen Species
Telomerase

Figure

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  Fig. 1 Identification of ADSCs. Expression of CD10, CD73, CD90, CD 105, CD34 and CD45 by ADSCs at passage 4. Numbers indicate the percentages of cells positive for each of the surface markers.

• 

  Fig. 2 Two lineage differentiation of ADSCs. (Left) Osteogenic differentiation and cell aggregates (were stained with alizarinred staining). Arrows show some of the mineralized cell aggregates (bar=200 μm). (Right) Differentiation into adipose cells. Arrows show lipid vacuoles generated after adipose differentiation (bar=200 μm).

• 

  Fig. 3 Growth and proliferation rates of adipose tissue-derived hMSCs in the presence of different concentrations of LC for 24, 48 and 72 hours, using MTT assay (*p<0.05 compared with control group).

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  Fig. 4 Relative hTERT gene expression levels of adipose tissue-derived hMSCs in the presence of different concentration of LC for 48 hours of incubation (*p<0.05 and **p<0.0001 compared with the control group). (Right) Absolute telomere length measurement of tissue-derived hMSCs in the presence of different concentration of LC for 48 hours of incubation (*p<0.05 and **p<0.001 compared with the control group).

Reference

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