Effect of long-term cultivation on biological characteristics of human adipose tissue-derived stem cells

Danišovič, Ľuboš; Boháč, Martin; Polák, Štefan; Žiaran, Stanislav; Oravcová, Lenka; Kuniaková, Marcela

doi:10.5176/2345-7821_acmr13.08

Cited by 21 publications

(28 citation statements)

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“…ASCs in long-term culture were shown to exhibit increased proliferation indicated by the increased percentage of cells in S and G2/M phases, which decreased in later passages. 20 Our study further revealed a relatively higher G2/M ratio in FGF2-treated ACSs at P5 and P10 relative to the control ASCs, and the proportion of G2/M phase ASCs in the FGF2-treated group became significantly lower than that in the control group at P17.…”

Section: Discussionsupporting

confidence: 64%

“…A common concern regarding the use of stem cell therapies is the risk of tumorigenicity . Previous studies have shown that in vitro‐expanded ASCs exhibited a low level of telomerase activity after passage 10, without evidence of alternative mechanisms for telomere lengthening . Hence, ASCs were considered to exhibit a low risk of tumorigenicity up to P20 .…”

Section: Discussionmentioning

confidence: 99%

“…FGF2 supplement in prolonged culture of ASC possibly alters not only the proliferative, self‐renewal ability, but also the genetic and epigenetic changes related to apoptosis, DNA damage repair, or tumorigenicity. In a study conducting long‐term ASC culture, elevated expression of tumor suppression gene p53 with decreased CDK1 (required for entry into M phase) and Bcl‐2 (anti‐apoptotic gene) expression in later passages was noted, indicating that ASCs might undergo apoptosis to inhibit malignant transformation . According to Gharibi et al, FGF2 supplement upregulated stemness gene Oct4 only at initial passages of ASC culture, and increased expression of senescence gene p16 and DNA repair gene Pold3 was noted at later passages .…”

Section: Discussionmentioning

confidence: 99%

“…Several studies have demonstrated the stability of human ASCs during prolonged cultivation with a low risk of tumorigenicity up to passage 20 . Although rare, spontaneous tumorigenic transformation of MSCs that are expanded in vitro has been reported, particularly when they were treated with certain carcinogens .…”

Section: Introductionmentioning

confidence: 99%

“…Several studies have demonstrated the stability of human ASCs during prolonged cultivation with a low risk of tumorigenicity up to passage 20. 10,20 Although rare, spontaneous tumorigenic transformation of MSCs that are expanded in vitro has been reported, particularly when they were treated with certain carcinogens. 21,22 For example, supplementing FGF2 in the culture medium of human bone marrow-derived MSCs transfected with TERT (telomerase reverse transcriptase) resulted in an increased potential for neoplastic transformation.…”

Section: Introductionmentioning

confidence: 99%

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The influence of fibroblast growth factor 2 on the senescence of human adipose-derived mesenchymal stem cells during long-term culture

Cheng

Lin

Young

et al. 2019

Stem Cells Translational Medicine

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Adipose‐derived mesenchymal stem cells (ASCs) exhibit great potential in regenerative medicine, and in vitro expansion is frequently necessary to obtain a sufficient number of ASCs for clinical use. Fibroblast growth factor 2 (FGF2) is a common supplement in the ASC culture medium to enhance cell proliferation. To achieve clinical applicability of ASC‐based products, prolonged culture of ASCs is sometimes required to obtain sufficient quantity of ASCs. However, the effect of FGF2 on ASCs during prolonged culture has not been previously determined. In this study, ASCs were subjected to prolonged in vitro culture with or without FGF2. FGF2 maintained the small cell morphology and expedited proliferation kinetics in early ASC passages. After prolonged in vitro expansion, FGF2‐treated ASCs exhibited increased cell size, arrested cell proliferation, and increased cellular senescence relative to the control ASCs. We observed an upregulation of FGFR1c and enhanced expression of downstream STAT3 in the initial passages of FGF2‐treated ASCs. The application of an FGFR1 or STAT3 inhibitor effectively blocked the enhanced proliferation of ASCs induced by FGF2 treatment. FGFR1c upregulation and enhanced STAT3 expression were lost in the later passages of FGF2‐treated ASCs, suggesting that the continuous stimulation of FGF2 becomes ineffective because of the refractory downstream FGFR1 and the STAT3 signaling pathway. In addition, no evidence of tumorigenicity was noted in vitro and in vivo after prolonged expansion of FGF2‐cultured ASCs. Our data indicate that ASCs have evolved a STAT3‐dependent response to continuous FGF2 stimulation which promotes the initial expansion but limits their long‐term proliferation.

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Section: Discussionsupporting

confidence: 64%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The influence of fibroblast growth factor 2 on the senescence of human adipose-derived mesenchymal stem cells during long-term culture

Cheng

Lin

Young

et al. 2019

Stem Cells Translational Medicine

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A Multilevel Approach to the Causes of Genetic Instability in Stem Cells

González

2022

Handbook of Stem Cell Therapy

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This chapter addresses the genetic instability in stem cells, a central feature that is an important determinant for the safety and effectiveness of cell-based therapies. First, DNA damage response mechanism and the gene network that regulates the DNA integrity homeostasis are revisited, focusing on relationship between genetic integrity and stemness maintenance. Also, several factors have been included that influence genetic instability in vivo, i.e., ROS generation and inflammation, and in vitro regarding cell isolation, culture conditions, i.e., oxygen levels and the use of feeder layers and different carriers, splitting procedures, passage number, etc. Telomere shortening, replicative senescence aneuploidy, and the senescence-associated secretory phenotype are different processes involved in deterioration of stem cells abilities for homing, reparability, and paracrine modulation. Moreover, less effective DNA repair upon senescence increases the propensity of developing tumors for stem cells that is one the main concerns related to genetic instability in stem cells. Nuclear organization and their determinants linked to chromosome aberrations and aneuploidy in stem cells and those epigenetic changes affecting stability are addressed. Differences between adipose, embryonic, and induced pluripotent stem cells are analyzed regarding to their ontogeny, changes in culture, and variation in proliferative capacity and stemness. The effect of reprogramming methods in genetic instability and variation in mutagenicity according to genes utilized for pluripotency induction, i.e., Yamanaka's factors and others, is discussed. Donor-related factors, i.e., age, smoking, and alcohol consumption, comorbidities, i.e., obesity, insulin resistance, diabetes, are mentioned for wide evaluation of genetic instability. The next years must witness consensus protocols that will contribute to control the effects of factors that alter stem cell genetic stability to increase the security and applicability of cell-based therapy.

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NAD+/NADH redox alterations reconfigure metabolism and rejuvenate senescent human mesenchymal stem cells in vitro

et al. 2020

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Human mesenchymal stem cells (hMSCs) promote endogenous tissue regeneration and have become a promising candidate for cell therapy. However, in vitro culture expansion of hMSCs induces a rapid decline of stem cell properties through replicative senescence. Here, we characterize metabolic profiles of hMSCs during expansion. We show that alterations of cellular nicotinamide adenine dinucleotide (NAD + /NADH) redox balance and activity of the Sirtuin (Sirt) family enzymes regulate cellular senescence of hMSCs. Treatment with NAD + precursor nicotinamide increases the intracellular NAD + level and re-balances the NAD + /NADH ratio, with enhanced Sirt-1 activity in hMSCs at high passage, partially restores mitochondrial fitness and rejuvenates senescent hMSCs. By contrast, human fibroblasts exhibit limited senescence as their cellular NAD + /NADH balance is comparatively stable during expansion. These results indicate a potential metabolic and redox connection to replicative senescence in adult stem cells and identify NAD + as a metabolic regulator that distinguishes stem cells from mature cells. This study also suggests potential strategies to maintain cellular homeostasis of hMSCs in clinical applications.

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Effect of long-term cultivation on biological characteristics of human adipose tissue-derived stem cells

Cited by 21 publications

References 0 publications

The influence of fibroblast growth factor 2 on the senescence of human adipose-derived mesenchymal stem cells during long-term culture

The influence of fibroblast growth factor 2 on the senescence of human adipose-derived mesenchymal stem cells during long-term culture

A Multilevel Approach to the Causes of Genetic Instability in Stem Cells

NAD+/NADH redox alterations reconfigure metabolism and rejuvenate senescent human mesenchymal stem cells in vitro

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