scholarly journals Epigenetic Conversion as a Safe and Simple Method to Obtain Insulin-secreting Cells from Adult Skin Fibroblasts

10.3791/53880 ◽  
2016 ◽  
Author(s):  
Tiziana A.L. Brevini ◽  
Georgia Pennarossa ◽  
Sara Maffei ◽  
Alessandro Zenobi ◽  
Fulvio Gandolfi
Keyword(s):  
Skin Fibroblasts ◽  
Simple Method ◽  
Insulin Secreting Cells ◽  
Epigenetic Conversion ◽  
Adult Skin

Epigenetic conversion of adult dog skin fibroblasts into insulin-secreting cells

2016 ◽  
Vol 211 ◽  
pp. 52-56 ◽  
Author(s):  
T.A.L. Brevini ◽  
G. Pennarossa ◽  
F. Acocella ◽  
S. Brizzola ◽  
A. Zenobi ◽  
...  
Keyword(s):  
Skin Fibroblasts ◽  
Insulin Secreting Cells ◽  
Epigenetic Conversion

Generation of Fluorescently Labeled Cell Lines, C3A Hepatoma Cells, and Human Adult Skin Fibroblasts to Study Coculture Models

2013 ◽  
Vol 37 (7) ◽  
pp. E123-E130 ◽  
Author(s):  
Anna Samluk ◽  
Karolina Ewa Zakrzewska ◽  
Krzysztof Dariusz Pluta
Keyword(s):  
Cell Lines ◽  
Hepatoma Cells ◽  
Skin Fibroblasts ◽  
Human Adult ◽  
Adult Skin

scholarly journals Quantitative Evaluation of the Mitochondrial DNA Depletion Syndrome

2010 ◽  
Vol 56 (7) ◽  
pp. 1119-1127 ◽  
Author(s):  
David Dimmock ◽  
Lin-Ya Tang ◽  
Eric S Schmitt ◽  
Lee-Jun C Wong
Keyword(s):  
Mitochondrial Dna ◽  
Real Time ◽  
Muscle Tissue ◽  
Skin Fibroblasts ◽  
Rapid Screening ◽  
Simple Method ◽  
Blood Samples ◽  
Mtdna Depletion ◽  
Coa Ligase ◽  
Mitochondrial Dna Depletion

Abstract Background: The mitochondrial DNA (mtDNA) depletion syndromes (MDDSs) are autosomal recessive disorders characterized by a reduction in cellular mtDNA content. Mutations in at least 9 genes [POLG, polymerase (DNA directed), gamma; DGUOK, deoxyguanosine kinase; TK2, thymidine kinase, mitochondrial; TYMP, thymidine phosphorylase; MPV17, MpV17 mitochondrial inner membrane protein; SUCLA2, succinate-CoA ligase, ADP-forming, beta subunit; SUCLG1, succinate-CoA ligase, alpha subunit; RRM2B, RRM2B, ribonucleotide reductase M2 B (TP53 inducible); and C10orf2, chromosome 10 open reading frame 2 (also known as TWINKLE)] have been reported to cause mtDNA depletion. In the clinical setting, a simple method to quantify mtDNA depletion would be useful before undertaking gene sequence analysis. Methods: Real-time quantitative PCR (qPCR) was used to measure the mtDNA content in blood, muscle, and liver samples and in skin fibroblast cultures from individuals suspected of mitochondrial disorders, with or without deleterious mutations in genes responsible for MDDS. Results: The mtDNA content was quantified in 776 tissue samples (blood, n = 341; muscle, n = 325; liver, n = 63; skin fibroblasts, n = 47) from control individuals. mtDNA content increased with age in muscle tissue, decreased with age in blood samples, and appeared to be unaffected by age in liver samples. In 165 samples (blood, n = 122; muscle, n = 21; liver, n = 15; skin fibroblasts, n = 7) from patients with molecularly proven MDDSs, severe mtDNA depletion was detected in liver and muscle tissue with high specificity and sensitivity. Blood samples were specific but not sensitive for detecting mtDNA depletion, and skin fibroblasts were not valuable for evaluating mtDNA depletion. Mutations in the POLG, RRM2B, and MPV17 genes were prospectively identified in 1 blood, 1 liver, and 3 muscle samples. Conclusions: Muscle and liver tissues, but not blood or skin fibroblasts, are potentially useful for rapid screening for mtDNA depletion with real-time qPCR.

Inter- and intra-site heterogeneity in the expression of fetal-like phenotypic characteristics by gingival fibroblasts: potential significance for wound healing

1994 ◽  
Vol 107 (5) ◽  
pp. 1333-1346 ◽  
Author(s):  
C.R. Irwin ◽  
M. Picardo ◽  
I. Ellis ◽  
P. Sloan ◽  
A. Grey ◽  
...  
Keyword(s):  
Wound Healing ◽  
Skin Fibroblasts ◽  
Gingival Fibroblast ◽  
Gingival Fibroblasts ◽  
Early Passage ◽  
Subsequent Work ◽  
Skin Cells ◽  
Fetal Fibroblasts ◽  
Adult Skin ◽  
Migratory Phenotype

We have previously reported that fetal and adult skin fibroblasts display distinctive migratory phenotypes on 3-D collagen substrata and that these behavioural characteristics may be quantified by a function defined as the cell density migration index (CDMI). Subsequent work indicated that this difference in migratory phenotype was due to the production by fetal fibroblasts of a migration stimulating factor (MSF) that is not produced by normal adult skin fibroblasts. We now present data indicating that: (a) unselected fibroblasts obtained from 14/14 (100%) of adult gingival explants expressed fetal-like CDMI values compared to only 1/10 (10%) of similarly explanted paired skin cells; (b) 12/12 (100%) of these gingival fibroblast lines also produced detectable quantities of MSF compared to 0/9 (0%) of the tested skin cells; (c) by microdissection studies, gingival fibroblasts obtained from different anatomical microdomains consisted of behaviourally distinct subpopulations, with cells derived from the papillary tips (PAP fibroblasts) displaying fetal-like CDMI values and persistent MSF production, whilst cells obtained from the deeper reticular tissue (RET fibroblasts) were adult-like with respect to these two criteria; (d) PAP fibroblasts were also smaller and achieved higher saturation cell densities compared to paired RET cells; (e) PAP fibroblasts passaged in vitro underwent a fetal-to-adult phenotypic transition characterized by the adoption of various RET cell characteristics, including the acquisition of CDMI values falling within the adult range and cessation in MSF production; and (f) early passage PAP fibroblasts incubated in the presence of an affinity-purified anti-MSF rabbit polyclonal antibody were induced to alter their migratory phenotype and exhibited CDMI values falling within the adult range. Statistical analysis indicated a highly significant correlation between the expression of a fetal-like CDMI and production of MSF (P < 0.00001, using the Fisher exact contingency test). Taken together, these observations suggest that the production of MSF by PAP fibroblasts is responsible for their characteristically fetal-like migratory behaviour. The existence of such inter- and intra-site phenotypic heterogeneity in populations of skin and gingival fibroblasts is discussed in the context of fibroblast lineage relationships and the possible contribution of persistently fetal-like fibroblast subpopulations to connective tissue function in wound healing.

Direct Conversion of Adult Skin Fibroblasts to Endothelial Cells by Defined Factors

Circulation ◽  
2014 ◽  
Vol 130 (14) ◽  
pp. 1168-1178 ◽  
Author(s):  
Jung-Kyu Han ◽  
Sung-Hwan Chang ◽  
Hyun-Ju Cho ◽  
Saet-Byeol Choi ◽  
Hyo-Suk Ahn ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Direct Conversion ◽  
Skin Fibroblasts ◽  
Adult Skin

scholarly journals Brief demethylation step allows the conversion of adult human skin fibroblasts into insulin-secreting cells

2013 ◽  
Vol 110 (22) ◽  
pp. 8948-8953 ◽  
Author(s):  
G. Pennarossa ◽  
S. Maffei ◽  
M. Campagnol ◽  
L. Tarantini ◽  
F. Gandolfi ◽  
...  
Keyword(s):  
Human Skin ◽  
Skin Fibroblasts ◽  
Human Skin Fibroblasts ◽  
Adult Human ◽  
Insulin Secreting Cells ◽  
Adult Human Skin
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