scholarly journals Rapid isolation and expansion of skin-derived precursor cells from human primary fibroblast cultures

Biology Open ◽  
2017 ◽  
Vol 6 (11) ◽  
pp. 1745-1755 ◽  
Author(s):  
Leithe Budel ◽  
Karima Djabali
Keyword(s):  
Precursor Cells ◽  
Primary Fibroblast ◽  
Rapid Isolation ◽  
Fibroblast Cultures ◽  
Human Primary Fibroblast

Expression of the familial Mediterranean fever gene and activity of the C5a inhibitor in human primary fibroblast cultures

Blood ◽  
2000 ◽  
Vol 96 (2) ◽  
pp. 727-731 ◽  
Author(s):  
Yaacov Matzner ◽  
Suzan Abedat ◽  
Eli Shapiro ◽  
Shlomit Eisenberg ◽  
Ariela Bar-Gil-Shitrit ◽  
...  
Keyword(s):  
Familial Mediterranean Fever ◽  
Human Fibroblasts ◽  
Skin Fibroblasts ◽  
Sterile Inflammation ◽  
Inhibitor Activity ◽  
Primary Fibroblast ◽  
Fibroblast Cultures ◽  
M694v Mutation ◽  
Human Primary Fibroblast ◽  
Mediterranean Fever

Abstract Familial Mediterranean fever (FMF) is an inherited disease whose manifestations are acute but reversible attacks of sterile inflammation affecting synovial and serosal spaces. The FMF gene (MEFV) was recently cloned, and it codes for a protein (pyrin/marenostrin) homologous to known nuclear factors. We previously reported the deficient activity of a C5a/interleukin (IL)–8 inhibitor, a physiologic regulator of inflammatory processes, in FMF serosal and synovial fluids. We now describe the concomitant expression ofMEFV and C5a/IL-8–inhibitor activity in primary cultures of human fibroblasts. Fibroblasts grown from synovial and peritoneal tissues displayed C5a/IL-8–inhibitor activity that could be further induced with phorbol myristate acetate (PMA) and IL-1β. Very low levels of chemotactic inhibitor were evident in skin fibroblast cultures or in peritoneal and skin fibroblasts obtained from FMF patients. MEFV was expressed in peritoneal and skin fibroblasts at a lower level than in neutrophils and could be further induced by PMA and IL-1β. In the FMF cultures, the MEFV transcript carried the M694V mutation, consistent with the genetic defect found in patients with this disease. MEFV was also expressed in other cell lines that do not produce C5a/IL-8 inhibitor. These findings suggest that human primary fibroblast cultures express MEFV and produce C5a/IL-8–inhibitor activity. The interrelationship between pyrin, the MEFV product, and the C5a/IL-8 inhibitor requires further investigation.

Expression of the familial Mediterranean fever gene and activity of the C5a inhibitor in human primary fibroblast cultures

Blood ◽  
2000 ◽  
Vol 96 (2) ◽  
pp. 727-731 ◽  
Author(s):  
Yaacov Matzner ◽  
Suzan Abedat ◽  
Eli Shapiro ◽  
Shlomit Eisenberg ◽  
Ariela Bar-Gil-Shitrit ◽  
...  
Keyword(s):  
Familial Mediterranean Fever ◽  
Human Fibroblasts ◽  
Skin Fibroblasts ◽  
Sterile Inflammation ◽  
Inhibitor Activity ◽  
Primary Fibroblast ◽  
Fibroblast Cultures ◽  
M694v Mutation ◽  
Human Primary Fibroblast ◽  
Mediterranean Fever

Familial Mediterranean fever (FMF) is an inherited disease whose manifestations are acute but reversible attacks of sterile inflammation affecting synovial and serosal spaces. The FMF gene (MEFV) was recently cloned, and it codes for a protein (pyrin/marenostrin) homologous to known nuclear factors. We previously reported the deficient activity of a C5a/interleukin (IL)–8 inhibitor, a physiologic regulator of inflammatory processes, in FMF serosal and synovial fluids. We now describe the concomitant expression ofMEFV and C5a/IL-8–inhibitor activity in primary cultures of human fibroblasts. Fibroblasts grown from synovial and peritoneal tissues displayed C5a/IL-8–inhibitor activity that could be further induced with phorbol myristate acetate (PMA) and IL-1β. Very low levels of chemotactic inhibitor were evident in skin fibroblast cultures or in peritoneal and skin fibroblasts obtained from FMF patients. MEFV was expressed in peritoneal and skin fibroblasts at a lower level than in neutrophils and could be further induced by PMA and IL-1β. In the FMF cultures, the MEFV transcript carried the M694V mutation, consistent with the genetic defect found in patients with this disease. MEFV was also expressed in other cell lines that do not produce C5a/IL-8 inhibitor. These findings suggest that human primary fibroblast cultures express MEFV and produce C5a/IL-8–inhibitor activity. The interrelationship between pyrin, the MEFV product, and the C5a/IL-8 inhibitor requires further investigation.

scholarly journals Effect of spermine synthase deficiency on polyamine biosynthesis and content in mice and embryonic fibroblasts, and the sensitivity of fibroblasts to 1,3-bis-(2-chloroethyl)-N-nitrosourea

2000 ◽  
Vol 351 (2) ◽  
pp. 439-447 ◽  
Author(s):  
Caroline A. MACKINTOSH ◽  
Anthony E. PEGG
Keyword(s):  
The Body ◽  
Normal Male ◽  
Male Mice ◽  
Embryonic Fibroblasts ◽  
Primary Fibroblast ◽  
Polyamine Biosynthesis ◽  
Spermine Synthase ◽  
Fibroblast Cultures ◽  
Phex Gene ◽  
Weaning Age

Mutant Gy male mice, which have previously been described as having disruption of the phosphate-regulating Phex gene and a spermine synthase gene [Meyer, Henley, Meyer, Morgan, McDonald, Mills and Price (1998) Genomics, 48, 289–295; Lorenz, Francis, Gempel, Böddrich, Josten, Schmahl and Schmidt (1998) Hum. Mol. Genet. 7, 541–547], as well as mutant Hyp male mice, which have disruption of the Phex gene only, were examined along with their respective normal male littermates. Biochemical analyses of extracts of brains, hearts and livers of 5-week-old mice showed that Gy males lacked any significant spermine synthase activity as well as spermine content. Organs of Gy males had a higher spermidine content. This was caused not only by the lack of conversion of spermidine into spermine, but also because of compensatory increases in the activities of other polyamine biosynthetic enzymes. Gy males were half the body weight of their normal male littermates at weaning age. Hyp males, however, were no different in size when compared with their controls. High mortality of Gy males occurs by weaning age and this mortality was shown to be largely post-natal. Embryonic fibroblasts were isolated from Gy males and their normal male littermates and were similarly shown to lack any significant spermine synthase activity as well as spermine content. The lack of spermine, however, had no significant effect on the growth of immortalized fibroblasts or of primary fibroblast cultures. Similarly, there was no difference in the time of senescence of primary fibroblast cultures from Gy males compared with cultures derived from normal male littermates. However, the lack of spermine did increase the sensitivity of immortalized fibroblasts to killing by the chloroethylating agent 1,3-bis(2-chloroethyl)-N-nitrosourea. Therefore both the Gy male mice and derived embryonic fibroblasts provide valuable models to study the importance of spermine and spermine synthase, without the use of inhibitors which may have additional side effects.

Cytokine-mediated PGE2expression in human colonic fibroblasts

1998 ◽  
Vol 275 (4) ◽  
pp. C988-C994 ◽  
Author(s):  
Edward C. Kim ◽  
Yingting Zhu ◽  
Valerie Andersen ◽  
Daniela Sciaky ◽  
H. James Cao ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Cell Lines ◽  
Shape Change ◽  
Cell Types ◽  
Mrna Levels ◽  
Primary Fibroblast ◽  
Fibroblast Cultures ◽  
Epithelial Cell Lines ◽  
Factor Α

We investigated prostanoid biogenesis in human colonic fibroblasts (CCD-18Co and 5 primary fibroblast cultures) and epithelial cell lines (NCM460, T84, HT-29, and LS 174T) and the effect of PGE2 on fibroblast morphology. Cytokine-stimulated PGE2production was measured. PGH synthase-1 and -2 (PGHS-1 and -2) protein and mRNA expression were evaluated. Basal PGE2 levels were low in all cell types (0.15–6.47 ng/mg protein). Treatment for 24 h with interleukin-1β (IL-1β; 10 ng/ml) or tumor necrosis factor-α (50 ng/ml), respectively, elicited maximal 25- and 6-fold inductions of PGE2 synthesis in CCD-18Co cultures and similar results in primary fibroblast cultures; maximal inductions with IL-1β in colonic epithelial cell lines were from zero to fivefold. Treatment of CCD-18Co fibroblasts with IL-1β caused maximal 21- and 53-fold increases, respectively, in PGHS-2 protein and mRNA levels without altering PGHS-1 expression. PGE2 (0.1 μmol/l) elicited a dramatic shape change in selected fibroblasts. Colonic fibroblasts are potentially important as cytokine targets and a source of and target for colonic prostanoids in vivo.

scholarly journals A dynamical systems model for the measurement of cellular senescence

2019 ◽  
Vol 16 (159) ◽  
pp. 20190311 ◽  
Author(s):  
Daniel Galvis ◽  
Darren Walsh ◽  
Lorna W. Harries ◽  
Eva Latorre ◽  
James Rankin
Keyword(s):  
Dynamical Systems ◽  
Replicative Senescence ◽  
Population Doubling ◽  
High Temporal Resolution ◽  
Model Parameters ◽  
Ki 67 ◽  
Primary Fibroblast ◽  
Systems Model ◽  
Human Primary Fibroblast

Senescent cells provide a good in vitro model to study ageing. However, cultures of ‘senescent’ cells consist of a mix of cell subtypes (proliferative, senescent, growth-arrested and apoptotic). Determining the proportion of senescent cells is crucial for studying ageing and developing new anti-degenerative therapies. Commonly used markers such as doubling population, senescence-associated β-galactosidase, Ki-67, γH2AX and TUNEL assays capture diverse and overlapping cellular populations and are not purely specific to senescence. A newly developed dynamical systems model follows the transition of an initial culture to senescence tracking population doubling, and the proportion of cells in proliferating, growth-arrested, apoptotic and senescent states. Our model provides a parsimonious description of transitions between these states accruing towards a predominantly senescent population. Using a genetic algorithm, these model parameters are well constrained by an in vitro human primary fibroblast dataset recording five markers at 16 time points. The computational model accurately fits to the data and translates these joint markers into the first complete description of the proportion of cells in different states over the lifetime. The high temporal resolution of the dataset demonstrates the efficacy of strategies for reconstructing the trajectory towards replicative senescence with a minimal number of experimental recordings.

Primary fibroblast cultures and karyotype analysis for the olive ridley sea turtle (Lepidochelys olivacea)

2013 ◽  
Vol 50 (5) ◽  
pp. 381-383 ◽  
Author(s):  
Tomokazu Fukuda ◽  
Masafumi Katayama ◽  
Kodzue Kinoshita ◽  
Takashi Kasugai ◽  
Hitoshi Okamoto ◽  
...  
Keyword(s):  
Karyotype Analysis ◽  
Sea Turtle ◽  
Olive Ridley ◽  
Primary Fibroblast ◽  
Lepidochelys Olivacea ◽  
Fibroblast Cultures

scholarly journals Biased Allelic Expression in Human Primary Fibroblast Single Cells

2015 ◽  
Vol 96 (1) ◽  
pp. 70-80 ◽  
Author(s):  
Christelle Borel ◽  
Pedro G. Ferreira ◽  
Federico Santoni ◽  
Olivier Delaneau ◽  
Alexandre Fort ◽  
...  
Keyword(s):  
Single Cells ◽  
Allelic Expression ◽  
Primary Fibroblast ◽  
Human Primary Fibroblast

scholarly journals Grammatophyllum speciosum Ethanolic Extract Promotes Wound Healing in Human Primary Fibroblast Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-6
Author(s):  
Saraporn Harikarnpakdee ◽  
Verisa Chowjarean
Keyword(s):  
Wound Healing ◽  
Ethanolic Extract ◽  
Skin Wound ◽  
Total Phenolic ◽  
Fibroblast Cells ◽  
Skin Wound Healing ◽  
Primary Fibroblast ◽  
Human Primary Fibroblast ◽  
Phytochemical Compounds

Grammatophyllum speciosum is a plant in Orchidaceae family which contains a variety of phytochemical compounds that might be beneficial for medicinal use. This study aimed to evaluate the activity of pseudobulb of G. speciosum extract (GSE) in wound healing processes in human primary fibroblast cells along with in vitro antioxidant activity and total phenolic content of GSE. Scratch wound healing assay indicated that GSE was capable of increasing migration rate after 6 and 9 hours of treatment. Besides, the extract was able to scavenge DPPH, ABTS, and superoxide anion radicals indicating the antioxidative property of GSE. This study suggested a novel role of the of pseudobulb extract of G. speciosum as a wound healing enhancer. The results from this study might be beneficial for the development of further novel active compounds for skin wound healing.

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