Induced pluripotent stem cells’ self-renewal and pluripotency is maintained by a bovine granulosa cell line-conditioned medium

Solari, Claudia; Losino, Noelia; Luzzani, Carlos; Waisman, Ariel; Blüguermann, Carolina; Questa, María; Sevlever, Gustavo; Miriuka, Santiago Gabriel; Barañao, Lino; Guberman, Alejandra

doi:10.1016/j.bbrc.2011.05.126

Cited by 7 publications

(7 citation statements)

References 32 publications

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“…To identify genes that are differentially expressed in undifferentiated compared to differentiated state, we studied their expression in three pluripotent stem cell lines: two ESC lines and an iPSC line generated by us, iPSC-20 [ 16 ]. We used two different mouse ESC lines, R1 and Ainv15, which were derived from embryos of different strains.…”

Section: Resultsmentioning

confidence: 99%

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Manganese Superoxide Dismutase Gene Expression Is Induced by Nanog and Oct4, Essential Pluripotent Stem Cells’ Transcription Factors

et al. 2015

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Pluripotent stem cells possess complex systems that protect them from oxidative stress and ensure genomic stability, vital for their role in development. Even though it has been reported that antioxidant activity diminishes along stem cell differentiation, little is known about the transcriptional regulation of the involved genes. The reported modulation of some of these genes led us to hypothesize that some of them could be regulated by the transcription factors critical for self-renewal and pluripotency in embryonic stem cells (ESCs) and in induced pluripotent stem cells (iPSCs). In this work, we studied the expression profile of multiple genes involved in antioxidant defense systems in both ESCs and iPSCs. We found that Manganese superoxide dismutase gene (Mn-Sod/Sod2) was repressed during diverse differentiation protocols showing an expression pattern similar to Nanog gene. Moreover, Sod2 promoter activity was induced by Oct4 and Nanog when we performed a transactivation assay using two different reporter constructions. Finally, we studied Sod2 gene regulation by modulating the expression of Oct4 and Nanog in ESCs by shRNAs and found that downregulation of any of them reduced Sod2 expression. Our results indicate that pluripotency transcription factors positively modulate Sod2 gene transcription.

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Section: Resultsmentioning

confidence: 99%

“…They were cultured and differentiated as previously described [ 13 – 15 ]. The iPSC-20 line was derived and validated previously by us [ 16 ], and cultured and differentiated as previously described [ 16 ]. This line was obtained by reprogramming day 13.5 mouse embryonic fibroblasts transduced with pHAGE-EF1a-STEMCCA [ 17 ].…”

Section: Methodsmentioning

confidence: 99%

Manganese Superoxide Dismutase Gene Expression Is Induced by Nanog and Oct4, Essential Pluripotent Stem Cells’ Transcription Factors

et al. 2015

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“…Induced pluripotent stem cells (iPSCs) are a type of pluripotent stem cells that can be generated directly from adult cells . iPSCs share the features of ESCs that are capable of self‐renewal and differentiation into three germ layers .…”

Section: Introductionmentioning

confidence: 99%

“…9 Induced pluripotent stem cells (iPSCs) are a type of pluripotent stem cells that can be generated directly from adult cells. [10][11][12] iPSCs share the features of ESCs that are capable of self-renewal and differentiation into three germ layers. 13 iPSCs can address immune rejection and ethical issues of the autologous cell transplantation.…”

Section: Introductionmentioning

confidence: 99%

Induced pluripotent stem cell‐derived conditional medium promotes Leydig cell anti‐apoptosis and proliferation via autophagy and Wnt/β‐catenin pathway

Chen

Hong

Chen

et al. 2018

J Cellular Molecular Medi

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Leydig cell transplantation is a better alternative in the treatment of androgen‐deficient males. The main purpose of this study was to investigate the effects of induced pluripotent stem cell‐derived conditioned medium (iPS‐CM) on the anti‐apoptosis, proliferation and function of immature Leydig cells (ILCs), and illuminate the underlying mechanisms. ILCs were exposed to 200 μmol/L hydrogen peroxide (H2O2) for 24 hours with or without iPS‐CM treatments. Cell apoptosis was detected by flow cytometric analysis. Cell proliferation was assessed using cell cycle assays and EdU staining. The steroidogenic enzyme expressions were quantified with Western blotting. The results showed that iPS‐CM significantly reduced H2O2‐induced ILC apoptosis through down‐regulation of autophagic and apoptotic proteins LC3‐I/II, Beclin‐1, P62, P53 and BAX as well as up‐regulation of BCL‐2, which could be inhibited by LY294002 (25 μmol/L). iPS‐CM could also promote ILC proliferation through up‐regulation of β‐catenin and its target proteins cyclin D1, c‐Myc and survivin, but was inhibited by XAV939 (10 μmol/L). The level of bFGF in iPS‐CM was higher than that of DMEM‐LG. Exogenous bFGF (20 ng/mL) or Wnt signalling agonist lithium chloride (LiCl) (20 mmol/L) added into DMEM‐LG could achieve the similar effects of iPS‐CM. Meanwhile, iPS‐CM could improve the medium testosterone levels and up‐regulation of LHCGR, SCARB1, STAR, CYP11A1, HSD3B1, CYP17A1, HSD17B3 and SF‐1 in H2O2‐induced ILCs. In conclusion, iPS‐CM could reduce H2O2‐induced ILC apoptosis through the activation of autophagy, promote proliferation through up‐regulation of Wnt/β‐catenin pathway and enhance testosterone production through increasing steroidogenic enzyme expressions, which might be used in regenerative medicine for future.

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“…We have recently shown that conditioned medium (CM) from a bovine granulosa cell line (BGC-CM) is able to maintain mouse pluripotent stem cells’ self-renewal, including ESC and induced pluripotent stem cells (iPSC), while preserving their unique properties, in culture without Leukemia Inhibitor Factor (LIF) addition [ 11 , 12 ]. Pluripotent stem cells growing on BGC-CM expressed stem cell markers and remained pluripotent.…”

Section: Introductionmentioning

confidence: 99%

EDA-Containing Fibronectin Increases Proliferation of Embryonic Stem Cells

et al. 2013

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Embryonic stem cells (ESC) need a set of specific factors to be propagated. They can also grow in conditioned medium (CM) derived from a bovine granulosa cell line BGC (BGC-CM), a medium that not only preserves their main features but also increases ESC´s proliferation rate. The mitogenic properties of this medium were previously reported, ascribing this effect to an alternative spliced generated fibronectin isoform that contains the extra domain A (FN EDA+). Here, we investigated if the FN EDA+ isoform increased proliferation of mouse and human ES cells. We analyzed cell proliferation using conditioned media produced by different mouse embryonic fibroblast (MEF) lines genetically engineered to express FN constitutively including or excluding the EDA domain (FN EDA-), and in media supplemented with recombinant peptides containing or not the EDA. We found that the presence of EDA in the medium increased mouse and human ESC’s proliferation rate. Here we showed for the first time that this FN isoform enhances ESC’s proliferation. These findings suggest a possible conserved behavior for regulation of ES cells proliferation by this FN isoform and could contribute to improve their culturing conditions both for research and cell therapy.

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Induced pluripotent stem cells’ self-renewal and pluripotency is maintained by a bovine granulosa cell line-conditioned medium

Cited by 7 publications

References 32 publications

Manganese Superoxide Dismutase Gene Expression Is Induced by Nanog and Oct4, Essential Pluripotent Stem Cells’ Transcription Factors

Manganese Superoxide Dismutase Gene Expression Is Induced by Nanog and Oct4, Essential Pluripotent Stem Cells’ Transcription Factors

Induced pluripotent stem cell‐derived conditional medium promotes Leydig cell anti‐apoptosis and proliferation via autophagy and Wnt/β‐catenin pathway

EDA-Containing Fibronectin Increases Proliferation of Embryonic Stem Cells

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