Transcriptome Profile of Human Fibroblasts in an <i>Ex Vivo</i> Culture

Rogińska, Dorota; Szumilas, Kamila; Zawiślak, Alicja; Wilk, Aleksandra; Stecewicz, Iwona; Brodkiewicz, Andrzej; Wiszniewska, Barbara

doi:10.7150/ijms.35693

Cited by 6 publications

(36 citation statements)

References 51 publications

(68 reference statements)

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“…The results presented in this paper continue on the gene expression profile analysis described by Machaliński et al 24 .…”

Section: Methodssupporting

confidence: 82%

“…A complete list of differentially expressed genes (DEGs), both upregulated and downregulated, in the cultured dermal fibroblasts from the male volunteer is presented in the supplementary materials of our previously published manuscript 24 . There, the DEGs of fibroblasts from the fist (C1), second (C2), and third (C3) cell passages were compared with those from the primary culture (C0).…”

Section: Resultsmentioning

confidence: 99%

“…Eca et al 2012 showed that reimplantation of autologous dermal fibroblasts cultured in medium supplemented with human serum was a viable method with no side effects 41 . This method is thus utilized to restore facial fullness and volume after autologous fibroblastic cell reimplantation in both sexes 22 , 24 , 42 , 43 .…”

Section: Discussionmentioning

confidence: 99%

“…In our previous study, we documented that autologous fibroblasts of the second passage, in contrast to cells from primary culture, were the optimal population for reimplantation into the dermis of male patients, because they had the highest secretory activity in releasing extracellular matrix components 24 . The present study is the continuation of our earlier analysis, in which we focused particularly on the expression of cell cycle-related genes in fibroblasts.…”

Section: Discussionmentioning

confidence: 99%

“…The detailed transcriptome profile analysis presented in our previous study 24 indicated that the second passage of fibroblasts is the optimal population due to the secretion and organization of extracellular matrix elements for potential autologous reimplantation to the skin. The present study continues this research with the comprehensive aim of examining the autologous fibroblast transcriptome profile by focusing on the processes of the cell cycle and cell proliferation status, in order to estimate the optimal passage of the tested cells with respect to reimplantation.…”

Section: Introductionmentioning

confidence: 98%

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Global Gene Expression of Cultured Human Dermal Fibroblasts: Focus on Cell Cycle and Proliferation Status in Improving the Condition of Face Skin

et al. 2021

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Chronological skin ageing is an inevitable physiological process that results in thin and sagging skin, fine wrinkles, and gradual dermal atrophy. The main therapeutic approaches to soft tissue augmentation involve using dermal fillers, where natural fillers, such as autologous fibroblasts, are involved in generating dermal matrix proteins. The aim of this study was to determine the global transcriptome profile of three passages of dermal autologous fibroblasts from a male volunteer, focusing on the processes of the cell cycle and cell proliferation status to estimate the optimal passage of the tested cells with respect to their reimplantation. We performed K-means clustering and validation of the expression of the selected mRNA by qRT-PCR. Ten genes were selected (ANLN, BUB1, CDC20, CCNA2, DLGAP5, MKI67, PLK1, PRC1, SPAG5, and TPX2) from the top five processes annotated to cluster 5. Detailed microarray analysis of the fibroblast genes indicated that the cell population of the third passage exhibited the highest number of upregulated genes involved in the cell cycle and cell proliferation. In all cases, the results of qRT-PCR confirmed the differences in expression of the selected mRNAs between fibroblasts from the primary culture (C0) and from the first (C1), second (C2), and third (C3) cell passage. Our results thus suggest that these cells might be useful for increasing fibroblast numbers after reimplantation into a recipient's skin, and the method used in this study seems to be an excellent tool for autologous transplantation allowing the rejuvenation of aging skin.

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“…The results presented in this paper continue on the gene expression profile analysis described by Machaliński et al 24 .…”

Section: Methodssupporting

confidence: 82%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

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Global Gene Expression of Cultured Human Dermal Fibroblasts: Focus on Cell Cycle and Proliferation Status in Improving the Condition of Face Skin

et al. 2021

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Transcriptomic and epigenetic assessment of ageing male skin identifies disruption of Ca²⁺ homeostasis; exacerbated by smoking and UV exposure

Pease

Wordsworth

Shanley

2022

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Skin ageing has been widely associated with the formation and presence of increasing quantities of senescent cells, the presence of which are thought to reduce cell renewal. This study aimed to identify key factors influencing fibroblast and skin aging in European males using RNA-seq data. Key differences in study designs included known sources of biological differences (sex, age, ethnicity), experimental differences, and environmental factors known to accelerate skin ageing (smoking, UV exposure) as well as study specific batch effects which complicated the analysis. To overcome these complications samples were stratified by these factors and differential expression assessed using Salmon and CuffDiff. Functional enrichment and consistency across studies, stratification's and tools identified age related alterations in the transcriptomes of fibroblasts and skin. Functional enrichment of results identified alterations in protein targeting to membranes and the ER, and altered calcium homeostasis in aged fibroblasts. Extension to skin controlled for differences in fibroblast culturing methods confirming transient age related alterations in intracellular calcium homeostasis. In middle aged males (40-65) increased keratinisation, skin, epithelial and epidermal development was seen in conjunction with alterations to ER Calcium uptake, leading to the identification of related processes including; an unfolded protein response, altered metabolism, increased MMP expression, and altered Calcium handling, which were further exacerbated by UV-exposure. Interestingly the genes and processes subsequently decreased in old males (> 65), which exhibited signs of increased senescence. Extension to Illumina 450k array data from ageing skin uncovered evidence of epigenetic regulation; genes and isoforms with overlapping differentially methylated CpGs were differentially expressed. Smoking led to additional enrichment of genes relating to tissue development, cell adhesion, vasculature development, peptide cross-linking, calcium homeostasis, cancer and senescence. The results consistently identified alterations in ER and golgi Calcium uptake, which disrupt intracellular and extracellular calcium gradients that regulate metabolic and differentiation signalling in skin and fibroblasts, leading to age related declines skin structure and function. Interestingly many diseases and infections with overlapping molecular consequences, (ER Calcium stress, reduced protein targeting to membranes) including COVID-19 are identified by the analysis, suggesting that COVID-19 infection compounds pre-existing cellular stresses in aged males, which could help explain higher COVID-19 mortality rates in aged males, as well as highlighting potential ways to reduce them.

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Transcriptomic and epigenetic assessment of ageing female skin and fibroblasts identifies age related reduced oxidative phosphorylation is exacerbated by smoking

Pease

Wordsworth

Shanley

2022

Preprint

View full text Add to dashboard Cite

Skin ageing has been widely associated with the formation and presence of increasing quantities of senescent cells, the presence of which are thought to reduce cell renewal. This study aimed to identify key factors influencing fibroblast and skin aging using RNA-seq data. Key differences in study designs included known sources of biological differences (sex, age, ethnicity), experimental differences, and environmental factors known to accelerate skin ageing (smoking, UV exposure) as well as study specific batch effects which complicated the analysis. To overcome these complications samples were stratified by these factors and differential expression assessed using Salmon and CuffDiff. Stratification of female fibroblast and skin samples combined with female specific normalisation of transcriptomic and methylation data sets increased functional enrichment and consistency across studies. The results identify the importance of considering environmental factors known to increase the rate of ageing (smoking status of donors, and UV-exposure status of skin and fibroblast samples) both independently and in combination for the identification of key ageing signatures. The results identified that in old (> 65) female skin decreases in the expression of transmembrane ion transporters coincide with increased methylation of oxidoreductases, and consequently reductions in respiration. This was further evidenced in old fibroblasts from smokers which identified reductions ion homeostasis, and the transcription of mitochondrial tRNAs, that were accompanied by reduced mitochondrial fission, reduced lipid catabolism and reduced immune signalling. These changes occurred in combination with reductions in cell proliferation, adhesion, ECM organisation, cell movement, cytoskeleton organisation and circulatory system development. Middle and old aged skin without environmental stratification's identified decreased expression of transmembrane ion transporters occurred alongside reductions in keratinisation, reduced mitochondrial fission, and this was associated with reduced metabolism (specifically carbohydrates), and consequently a reduction in the production of lipids (phospholipids for membranes and others) occured, exacerbating ion homeostasis issues at a keratinocyte level. Interestingly in skin the combined impacts of UV-exposure, smoking and ageing yielded different results, increased expression of calcium homeostasis genes, cell adhesion molecules (integrins), structural membrane constituents (loricrin, mucins, keratins and collagens), increased cornification, as well as structural cytoskeletal molecules (KRTAPs). This occurred alongside increased expression of genes involved in skin peeling (kalikriens), proliferation and differentiation, glycosylation, oxidative stress, autophagy, lactose metabolism, and lipid catabolism. Aged UV-exposed skin from smokers is on the whole more fibrous, with cells showing significant cell membrane and cytoskeletal structural changes, similar to those seen in skin cancers. Interestingly in non-UV-exposed skin from smokers most of these processes were reduced, and in within age group comparisons of smokers they were also reduced, suggesting that smoking reduced skin development and regeneration. Female specific analysis of smokers from different age groups enrichment results identified additional factors relating to tissue development, cell adhesion, vasculature development, peptide cross-linking, calcium homeostasis, cancer and senescence, leading to age related declines skin structure and function. Interestingly many diseases and infections with overlapping molecular consequences, (ER Calcium stress, reduced protein targeting to membranes) including human cytomegalovirus and herpes simplex virus are identified by the age only analysis, suggesting that viral infections and ageing have similar molecular consequences for cells.

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Transcriptome Profile of Human Fibroblasts in an Ex Vivo Culture

Cited by 6 publications

References 51 publications

Global Gene Expression of Cultured Human Dermal Fibroblasts: Focus on Cell Cycle and Proliferation Status in Improving the Condition of Face Skin

Global Gene Expression of Cultured Human Dermal Fibroblasts: Focus on Cell Cycle and Proliferation Status in Improving the Condition of Face Skin

Transcriptomic and epigenetic assessment of ageing male skin identifies disruption of Ca²⁺ homeostasis; exacerbated by smoking and UV exposure

Transcriptomic and epigenetic assessment of ageing female skin and fibroblasts identifies age related reduced oxidative phosphorylation is exacerbated by smoking

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Transcriptome Profile of Human Fibroblasts in an Ex Vivo Culture

Cited by 6 publications

References 51 publications

Global Gene Expression of Cultured Human Dermal Fibroblasts: Focus on Cell Cycle and Proliferation Status in Improving the Condition of Face Skin

Global Gene Expression of Cultured Human Dermal Fibroblasts: Focus on Cell Cycle and Proliferation Status in Improving the Condition of Face Skin

Transcriptomic and epigenetic assessment of ageing male skin identifies disruption of Ca2+ homeostasis; exacerbated by smoking and UV exposure

Transcriptomic and epigenetic assessment of ageing female skin and fibroblasts identifies age related reduced oxidative phosphorylation is exacerbated by smoking

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Transcriptomic and epigenetic assessment of ageing male skin identifies disruption of Ca²⁺ homeostasis; exacerbated by smoking and UV exposure