Proliferative potential and phenotypic analysis of long-term cultivated human granulosa cells initiated by addition of follicular fluid

There has been considerable interest in the generation of functional mesenchymal stromal cell (MSC) preparations from induced pluripotent stem cells (iPSCs) and this is now regarded as a potential source of unlimited, standardized, high‐quality cells for therapeutic applications in regenerative medicine. Although iMSCs meet minimal criteria for defining MSCs in terms of marker expression, there are substantial differences in terms of trilineage potential, specifically a marked reduction in chondrogenic and adipogenic propensity in iMSCs compared with bone marrow‐derived (BM) MSCs. To reveal the cellular basis underlying these differences, we conducted phenotypic, functional, and genetic comparisons between iMSCs and BM‐MSCs. We found that iMSCs express very high levels of both KDR and MSX2 compared with BM‐MSCs. In addition, BM‐MSCs had significantly higher levels of PDGFRα . These distinct gene expression profiles were maintained during culture expansion, suggesting that prepared iMSCs are more closely related to vascular progenitor cells (VPCs). Although VPCs can differentiate along the chondrogenic, osteogenic, and adipogenic pathways, they require different inductive conditions compared with BM‐MSCs. These observations suggest to us that iMSCs, based on current widely used preparation protocols, do not represent a true alternative to primary MSCs isolated from BM. Furthermore, this study highlights the fact that high levels of expression of typical MSC markers such as CD73, CD90, and CD105 are insufficient to distinguish MSCs from other mesodermal progenitors in differentiated induced pluripotent stem cell cultures. stem cells 2019;37:754–765

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“…Cell growth rate was determined as cumulative population doubling (CPD) .

CPD = \log ((), No . ([], harvesting) / No . ([], seeding)) / \log ((), 2)

…”

Section: Methodsmentioning

confidence: 99%

Induced Pluripotent Stem Cell-Derived Mesenchymal Stromal Cells Are Functionally and Genetically Different From Bone Marrow-Derived Mesenchymal Stromal Cells

Shaw

Murphy

et al. 2019

Stem Cells

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“…Population-doubling time was calculated using the following formula: number of hours from seeding to collection/((log n ( t ) − log n ( t 0) )/log 2 ). n (t) is the number of cells at time of passage and n (t 0 ) is the number of cells seeded at previous passage 32 .…”

Section: Methodsmentioning

confidence: 99%

XPO1-dependent nuclear export is a druggable vulnerability in KRAS-mutant lung cancer

Kim

McMillan

Kim

et al. 2016

Nature

177

159

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The common participation of oncogenic KRAS proteins in many of the most lethal human cancers, together with the ease of detecting somatic KRAS mutant alleles in patient samples, has spurred persistent and intensive efforts to develop drugs that inhibit KRAS activity1. However, advances have been hindered by the pervasive inter- and intra-lineage diversity in the targetable mechanisms that underlie KRAS-driven cancers, limited pharmacological accessibility of many candidate synthetic-lethal interactions and the swift emergence of unanticipated resistance mechanisms to otherwise effective targeted therapies. Here we demonstrate the acute and specific cell-autonomous addiction of KRAS-mutant non-small-cell lung cancer cells to receptor-dependent nuclear export. A multi-genomic, data-driven approach, utilizing 106 human non-small-cell lung cancer cell lines, was used to interrogate 4,725 biological processes with 39,760 short interfering RNA pools for those selectively required for the survival of KRAS-mutant cells that harbour a broad spectrum of phenotypic variation. Nuclear transport machinery was the sole process-level discriminator of statistical significance. Chemical perturbation of the nuclear export receptor XPO1 (also known as CRM1), with a clinically available drug, revealed a robust synthetic-lethal interaction with native or engineered oncogenic KRAS both in vitro and in vivo. The primary mechanism underpinning XPO1 inhibitor sensitivity was intolerance to the accumulation of nuclear IκBα (also known as NFKBIA), with consequent inhibition of NFκB transcription factor activity. Intrinsic resistance associated with concurrent FSTL5 mutations was detected and determined to be a consequence of YAP1 activation via a previously unappreciated FSTL5–Hippo pathway regulatory axis. This occurs in approximately 17% of KRAS-mutant lung cancers, and can be overcome with the co-administration of a YAP1–TEAD inhibitor. These findings indicate that clinically available XPO1 inhibitors are a promising therapeutic strategy for a considerable cohort of patients with lung cancer when coupled to genomics-guided patient selection and observation.

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“…Five unrelated lines of human granulosa cells (COV434, GC8, GC15, GC671, and GC672) were used and cultured as described by Bruckova and colleagues [23,24]. COV434 cell line represented cancerous cells originating in a granulosa cell tumor of ovary [25].…”

Section: Cell Culture and Harvesting Of Conditioned Mediamentioning

confidence: 99%

Presence of growth/differentiation factor-15 cytokine in human follicular fluid, granulosa cells, and oocytes

Souček

Malenovská²,

Kahounová

et al. 2018

J Assist Reprod Genet

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Proliferative potential and phenotypic analysis of long-term cultivated human granulosa cells initiated by addition of follicular fluid

Cited by 22 publications

References 43 publications

Induced Pluripotent Stem Cell-Derived Mesenchymal Stromal Cells Are Functionally and Genetically Different From Bone Marrow-Derived Mesenchymal Stromal Cells

Induced Pluripotent Stem Cell-Derived Mesenchymal Stromal Cells Are Functionally and Genetically Different From Bone Marrow-Derived Mesenchymal Stromal Cells

XPO1-dependent nuclear export is a druggable vulnerability in KRAS-mutant lung cancer

Presence of growth/differentiation factor-15 cytokine in human follicular fluid, granulosa cells, and oocytes

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