scholarly journals Evaluation of X-Inactivation Status and Cytogenetic Stability of Human Dermal Fibroblasts after Long-Term Culture

2010 ◽  
Vol 2010 ◽  
pp. 1-5
Author(s):  
Zhi-Gang Xue ◽  
Zhan-Ping Shi ◽  
Juan Dong ◽  
Ting-Ting Liao ◽  
Yan-Peng Wang ◽  
...  
Keyword(s):  
X Chromosome ◽  
Normal Karyotype ◽  
Biological Properties ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
X Chromosomes ◽  
High Passage ◽  
Primary Fibroblasts ◽  
Induced Pluripotent

Human primary fibroblasts are a popular type of somatic cells for the production of induced pluripotent stem (iPS) cells. Here we characterized biological properties of primary fibroblasts in terms of cell-growth rate, cytogenetic stability, and the number of inactive X chromosomes during long-term passaging. We produced eight lines of female human dermal fibroblasts (HDFs) and found normal karyotype and expected pattern of X chromosome inactivation (XCI) at low passages (Passage P1-5). However, four out of the eight HDF lines at high passage numbers (≥P10) exhibited duplicated hallmarks of inactive X chromosome including two punctuate signals of histone H3 lysine 27 trimethylation (H3K27me3) and X inactive-specific transcript (XIST) RNA signals in approximately 8.5–18.5% of the cells. Our data suggest that the copy number of inactive X chromosomes in a subset of female HDF is increased by a two-fold. Consistently, DNA fluorescent in situ hybridization (FISH) identified 3-4 copies of X chromosomes in one nucleus in this subset of cells with two inactive Xs. We conclude that female HDF cultures exhibit a higher risk of genetic anomalies such as carrying an increased number of X chromosomes including both active and inactive X chromosomes at a high passage (≥P10).

Postmitotic human dermal fibroblasts preserve intact feeder properties for epithelial cell growth after long-term cryopreservation

1990 ◽  
Vol 26 (7) ◽  
pp. 709-712 ◽  
Author(s):  
Alain Limat ◽  
Thomas Hunziker ◽  
Colette Boillat ◽  
Friedrich Noser ◽  
Ulrich Wiesmann
Keyword(s):  
Cell Growth ◽  
Epithelial Cell ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts

Long-term fate of silica nanoparticles interacting with human dermal fibroblasts

Biomaterials ◽  
2012 ◽  
Vol 33 (17) ◽  
pp. 4431-4442 ◽  
Author(s):  
Sandrine Quignard ◽  
Gervaise Mosser ◽  
Michel Boissière ◽  
Thibaud Coradin
Keyword(s):  
Silica Nanoparticles ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts

scholarly journals Puerarin blocks the aging phenotype in human dermal fibroblasts

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0249367
Author(s):  
Yuki Kamiya ◽  
Mao Odama ◽  
Aki Mizuguti ◽  
Shigeru Murakami ◽  
Takashi Ito
Keyword(s):  
Dermal Fibroblast ◽  
Smooth Muscle Actin ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
High Passage ◽  
Normal Human ◽  
Low Passage ◽  
Passage Cells ◽  
Beta Galactosidase ◽  
Functional Defects

Dermal fibroblast aging contributes to aging-associated functional defects in the skin since dermal fibroblasts maintain skin homeostasis by interacting with the epidermis and extracellular matrix. Here, we found that puerarin, an isoflavone present in Pueraria lobata (Kudzu), can prevent the development of the aging-phenotype in human dermal fibroblasts. Normal human dermal fibroblasts (NHDFs) were subcultivated and high-passage cells were selected as senescent cells, whereas low-passage cells were selected as a young cell control. Puerarin treatment increased cell proliferation and decreased the proportion of senescence-associated beta-galactosidase-positive cells in a high-passage culture of NHDFs. Moreover, puerarin treatment reduced the number of smooth muscle actin (SMA)-positive myofibroblasts and the expression of a reticular fibroblast marker, calponin 1 (CNN1), which were induced in high-passage NHDFs. Fulvestrant, an estrogen receptor antagonist, blocked the puerarin-mediated downregulation of SMA and CNN1. Our results suggest that puerarin may be a useful functional food that alleviates aging-related functional defects in dermal fibroblasts.

scholarly journals Puerarin blocks aging phenotype in cultured human dermal fibroblasts

2020 ◽  
Author(s):  
Yuki Kamiya ◽  
Mao Odama ◽  
Aki Mizuguti ◽  
Shigeru Murakami ◽  
Takashi Ito
Keyword(s):  
Dermal Fibroblast ◽  
Smooth Muscle Actin ◽  
Dermal Fibroblasts ◽  
Pueraria Lobata ◽  
Human Dermal Fibroblasts ◽  
Proliferating Cells ◽  
Food Factor ◽  
High Passage ◽  
Beta Galactosidase ◽  
Functional Defects

AbstractDermal fibroblast aging contributes to aging-associated functional defects in the skin since dermal fibroblasts are important to maintain skin homeostasis by interacting with epidermis and extracellular matrix. Here we identified that puerarin, an isoflavone contained in Pueraria lobata (Kudzu), can prevent the aging-phenotype of human dermal fibroblasts. Puerarin treatment increased in proliferating cells and decreased in senescence-associated beta-galactosidase positive cells in the high-passage culture of dermal fibroblasts. Moreover, puerarin reduced smooth muscle actin-positive myofibroblasts and the expression of a reticular fibroblast marker, calponin 1 (CNN1), which were induced in high-passage fibroblasts. Fulvestrant, an estrogen receptor antagonist, blocked puerarin-mediated downregulation of SMA and CNN1. Our results suggest that puerarin may be a useful food factor that alleviates aging-related functional defects in the skin.

scholarly journals Brief Report: Inhibition of miR-145 Enhances Reprogramming of Human Dermal Fibroblasts to Induced Pluripotent Stem Cells

Stem Cells ◽  
2015 ◽  
Vol 34 (1) ◽  
pp. 246-251 ◽  
Author(s):  
Tomas Barta ◽  
Lucie Peskova ◽  
Joseph Collin ◽  
David Montaner ◽  
Irina Neganova ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
Induced Pluripotent

scholarly journals Tailoring Gellan Gum Spongy-Like Hydrogels’ Microstructure by Controlling Freezing Parameters

Polymers ◽  
2020 ◽  
Vol 12 (2) ◽  
pp. 329 ◽  
Author(s):  
Helena R. Moreira ◽  
Lucília P. da Silva ◽  
Rui L. Reis ◽  
Alexandra P. Marques
Keyword(s):  
Biological Properties ◽  
Dermal Fibroblasts ◽  
Gellan Gum ◽  
Fine Tuning ◽  
Freezing Rate ◽  
Human Dermal Fibroblasts ◽  
Structure And Properties ◽  
Nucleation Time ◽  
Standard Process ◽  
Whole Process

Gellan gum (GG) spongy-like hydrogels have been explored for different tissue engineering (TE) applications owing to their highly attractive hydrogel-like features, and improved mechanical resilience and cell performance. Although the whole process for the preparation of these materials is well-defined, we hypothesized that variations occurring during the freezing step lead to batch-to-batch discrepancies. Aiming to address this issue, two freezing devices were tested, to prepare GG spongy-like hydrogels in a more reproducible way. The cooling and freezing rates, the nucleation time and temperature, and the end freezing time were determined at different freezing temperatures (−20, −80, and −210 °C). The efficacy of the devices was assessed by analyzing the physicochemical, mechanical, and biological properties of different formulations. The cooling rate and freezing rate varied between 0.1 and 128 °C/min, depending on the temperature used and the device. The properties of spongy-like hydrogels prepared with the tested devices showed lower standard deviation in comparison to those prepared with the standard process, due to the slower freezing rate of the hydrogels. However, with this method, mean pore size was significantly lower than that with the standard method. Cell entrapment, adhesion, and viability were not affected as demonstrated with human dermal fibroblasts. This work confirmed that batch-to-batch variations are mostly due to the freezing step and that the tested devices allow fine tuning of the scaffolds’ structure and properties.

Sign in / Sign up

Export Citation Format

Share Document