Determination of mTOR activity depending on donor age of mesenchymal stem cells isolated from adipose tissue

Year 2023, Volume: 16 Issue: 4, 704 - 714, 01.10.2023

Hayriye Sarıkaya , Elif Önder , Nazlı Çil , Ergun Mete , Gülçin Abban Mete

https://doi.org/10.31362/patd.1337120

Abstract

Purpose: Aging in living organisms is an inevitable physiological consequence. Cellular senescence occurs not
only in cells that have completed their differentiation, but also in stem cells. Mammalian target of Rapamycin
protein complex (mTOR) has an important role in cell growth and metabolism. The mTOR, which plays an
important role in cell proliferation, also regulates cellular aging and directs the bioenergetic infrastructure. The
aim of the study is to determine the mTOR expression of mesenchymal stem cell (MSC) obtained from adipose
tissue depending on the donor age.
Materials and methods: Six-week-old pubertal rats were named Group 1 (n=6), 10-12-week-old reproductive
period rats were named Group 2 (n=6), and 20-month-old rats were named Group 3 (n=6). The isolation of MSC
was performed by primary explant culture method from adipose tissue taken from groups. Characterization and
differentiation experiments were performed in MSC obtained. The activity of mTOR (mTORC1 and mTORC2)
in MSC was determined by qRT-PCR method. Caspase 3, 8, 9, Bax and Bcl-2 expressions were evaluated by
Real-time polymerase chain reaction (qRT-PCR) method.
Results: In our study, it was determined that the highest expression of apoptotic markers was in Group 1 and
the lowest expression was in Group 2. When mTOR expression was evaluated, mTORC1 was found to be
highest in Group 2 and lowest in Group 1. mTORC2 expression in Group 1 was lower than in other groups.
Although the expression of mTORC1 and mTORC2 in Group 3 was not as high as in Group 2, it was statistically
significant (p<0.05).
Conclusion: In this study, we found that both mTORC1 and mTORC2 are differentially expressed in stem cells
depending on donor age. Further studies are needed to better understand the functional consequences of this
difference.

Keywords

Aging, adipose tissue, mTOR, mesenchymal stem cells

Supporting Institution

This work was supported by The Pamukkale University Scientific Research Projects Coordination Unit

Project Number

Project number: 2021SABE003

Thanks

Thanks to The Pamukkale University Scientific Research Projects Coordination Unit for their supports.

Adipoz dokudan izole edilen mezenkimal kök hücrelerin donör yaşına bağlı olarak mTOR aktivitesinin belirlenmesi

Abstract

Amaç: Canlı organizmalarda yaşlanma kaçınılmaz bir fizyolojik sonuçtur. Hücresel yaşlanma sadece
farklılaşmasını tamamlamış hücrelerde değil, kök hücrelerde de meydana gelir. Rapamisin protein kompleksinin
memeli hedefi (mTOR), hücre büyümesinde ve metabolizmasında önemli bir role sahiptir. Hücre çoğalmasında
önemli rol oynayan mTOR, aynı zamanda hücresel yaşlanmayı düzenler ve biyoenerjetik altyapıyı yönlendirir.
Çalışmanın amacı, donör yaşına bağlı olarak yağ dokusundan elde edilen mezenkimal kök hücrelerin (MSC)
mTOR ekspresyonunu belirlemektir.
Gereç ve yöntem: Altı haftalık pubertal sıçanlar Grup 1 (n=6), 10-12 haftalık reprodüktif dönem sıçanları Grup 2
(n=6) ve 20 aylık sıçanlar Grup 3 (n=6) olarak isimlendirildi. Gruplardan alınan yağ dokusundan primer eksplant
kültür metodu ile MSC izolasyonu yapıldı. Elde edilen MSC'de karakterizasyon ve farklılaşma deneyleri yapıldı.
Kök hücrelerdeki mTOR aktivitesi (mTORC1 ve mTORC2) qRT-PCR yöntemi ile belirlendi. Caspase 3, 8, 9, Bax
ve Bcl-2 ekspresyonları Real-time polymerase chain reaction (qRT-PCR) yöntemi ile değerlendirildi.
Bulgular: Çalışmamızda apoptotik belirteçlerin en yüksek ekspresyonunun Grup 1'de, en düşük ekspresyonun
Grup 2'de olduğu belirlendi. mTOR ekspresyonu değerlendirildiğinde, mTORC1 en yüksek Grup 2'de, en düşük
Grup 1'de bulundu. Grup 1'deki mTORC2 ifadesi diğer gruplara göre daha düşüktü. Grup 3'teki mTORC1 ve
mTORC2 ekspresyonu Grup 2'deki kadar yüksek olmasa da istatistiksel olarak anlamlıydı (p<0.05).
Sonuç: Bu çalışmada hem mTORC1 hem de mTORC2'nin donör yaşına bağlı olarak kök hücrelerde farklı
şekilde eksprese edildiğini bulduk. Bu farkın fonksiyonel sonuçlarını daha iyi anlamak için daha fazla çalışmaya
ihtiyaç vardır.

Keywords

Yaşlanma, yağ doku, mTOR, mezenkimal kök hücreler

Project Number

Project number: 2021SABE003

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