Development of a chemically definedin vitroculture system to effectively stimulate the proliferation of adult human dermal fibroblasts

2015 ◽  
Vol 24 (7) ◽  
pp. 543-545 ◽  
Author(s):  
Min Seong Kim ◽  
Jung Im Yun ◽  
Seung Pyo Gong ◽  
Ji Yeon Ahn ◽  
Jeong Mook Lim ◽  
...  
Keyword(s):  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
Adult Human

scholarly journals Induction of functional platelets from mouse and human fibroblasts by p45NF-E2/Maf

Blood ◽  
2012 ◽  
Vol 120 (18) ◽  
pp. 3812-3821 ◽  
Author(s):  
Yukako Ono ◽  
Yuhuan Wang ◽  
Hidenori Suzuki ◽  
Shinichiro Okamoto ◽  
Yasuo Ikeda ◽  
...  
Keyword(s):  
Cultured Cells ◽  
Human Fibroblasts ◽  
Mouse Fibroblast ◽  
Dermal Fibroblasts ◽  
Induction Medium ◽  
Human Dermal Fibroblasts ◽  
3T3 Cells ◽  
Gene Expressions ◽  
Immunodeficient Mice ◽  
Adult Human

Abstract Determinant factors leading from stem cells to megakaryocytes (MKs) and subsequently platelets have yet to be identified. We now report that a combination of nuclear factor erythroid–derived 2 p45 unit (p45NF-E2), Maf G, and Maf K can convert mouse fibroblast 3T3 cells and adult human dermal fibroblasts into MKs. To screen MK-inducing factors, gene expressions were compared between 3T3 cells that do not differentiate into MKs and 3T3-L1 cells known to differentiate into MKs. 3T3 cells transfected with candidate factors were cultured in a defined MK lineage induction medium. Among the tested factors, transfection with p45NF-E2/MafG/MafK lead to the highest frequency of CD41-positive cells. Adult human dermal fibroblasts transfected with these genes were cultured in MK lineage induction medium. Cultured cells had megakaryocytic features, including surface markers, ploidy, and morphology. More than 90% of MK-sized cells expressed CD41, designated induced MK (iMK). Infusion of these iMK cells into immunodeficient mice led to a time-dependent appearance of CD41-positive, platelet-sized particles. Blood samples from iMK-infused into thrombocytopenic immunodeficient mice were perfused on a collagen-coated chip, and human CD41-positive platelets were incorporated into thrombi on the chip, demonstrating their functionality. These findings demonstrate that a combination of p45NF-E2, Maf G, and Maf K is a key determinant of both megakaryopoiesis and thrombopoiesis.

Tissue-protective effects of French maritime pine bark (Pycnogenol) on glutamate-induced cytotoxicity in adult human dermal fibroblasts

2017 ◽  
Vol 280 ◽  
pp. S159
Author(s):  
Çiğdem Sevim ◽  
Elif Doğan ◽  
Ali Taghizadehghalehjoughi ◽  
Semin Gedikli ◽  
Mustafa Özkaraca ◽  
...  
Keyword(s):  
Dermal Fibroblasts ◽  
Maritime Pine ◽  
Pine Bark ◽  
Protective Effects ◽  
Human Dermal Fibroblasts ◽  
Adult Human

scholarly journals Derivation of human differential photoreceptor cells from adult human dermal fibroblasts by defined combinations of CRX , RAX , OTX 2 and NEUROD

2014 ◽  
Vol 19 (3) ◽  
pp. 198-208 ◽  
Author(s):  
Yuko Seko ◽  
Noriyuki Azuma ◽  
Toshiyuki Ishii ◽  
Yukari Komuta ◽  
Kiyoko Miyamoto ◽  
...  
Keyword(s):  
Photoreceptor Cells ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
Adult Human

Induction of Megakaryocytes From Fibroblasts by p45NF-E2/Maf

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 908-908 ◽  
Author(s):  
Yuhuan Wang ◽  
Yukako Ono ◽  
Yasuo Ikeda ◽  
Shinichiro Okamoto ◽  
Mitsuru Murata ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Transcription Factors ◽  
Cell Line ◽  
Thrombus Formation ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
Flow Cytometry Analysis ◽  
3T3 Cells ◽  
Adult Human ◽  
Before And After

Abstract Abstract 908 The determinant factors leading from stem cells to megakaryocytes (MK) and the subsequent release of platelets have yet to be identified. Here we report that a combination of transcription factors p45NF-E2, MafG, and MafK converts mouse fibroblast cell line 3T3 and adult human dermal fibroblasts into MKs. We first performed screening for MK-inducing factors using the preadipocyte cell line 3T3-L1 known to be able to differentiate into MKs in a previously defined MK lineage induction media containing thrombopoietin (Zauli G et al, Blood 1997). Gene expression levels of candidate transcription factors were compared between 3T3 cells that do not differentiate into MKs and 3T3-L1 cells using quantitative real-time PCR. The expressions of p45NF-E2 and CEBPα were undetectable in 3T3 cells, whereas expression was seen in 3T3-L1 cells. Both cell lines had similar expression levels of GATA2, RUNX1, FOG1, and PPARγ and undetectable levels of GATA1. 3T3 cells transfected with p45NF-E2, its binding protein Maf, or CEBPα, and 3T3 cells transfected with any combination of these genes, were cultured in MK lineage induction media for 8 days. MK differentiation was assessed using CD41 expression in MK-sized cells by flow cytometry analysis. Frequency of CD41 expression in the 3T3 cells varied based on transgenic factors expressed: 19 ± 12% (p45NF-E2); 25 ± 13% (p45NF-E2/MafG/MafK); 18 ± 3% (p45NF-E2/MafG/MafK/CEBPα); 13 ± 2% (p45NF-E2/CEBPα); and 4 ± 1% (CEBPα). These results suggest that p45NF-E2 with or without Maf are defined factors for reprogramming of 3T3 fibroblast to MKs. We then tested whether other adult non-hematopoietic tissues could be forced into megakaryopoiesis by ectopic expression of p45NF-E2 and Maf. Adult human dermal fibroblasts (Cell Applications) were transfected with p45NF-E2/Maf genes and cultured in MK lineage induction media for 12 days. Large-sized cells were isolated using a 2-step density BSA gradient. By flow cytometry analysis, >99% of these cells expressed CD41. DNA ploidy of the induced MKs (iMKs) ranged from 2N to 16N with a mean of 4N. We then infused 5 × 105 large-sized iMKs into NOG mouse after induction of mild thrombocytopenia in them at day 7 after irradiation with 2.0 Gy. Tail vein samples were obtained from the tested female recipient NOG mice before and after 5 min, 30 min, 90 min, and 3h post iMK infusion. Flow cytometry analysis was performed on each sample stained with PE-conjugated anti-mouse CD41 and FITC-conjugated anti-human CD41 antibody. Human CD41-positive platelet-sized cells in the sample from before and after 5 min, 30 min, 90 min, and 3h represented 0.9%, 0.5%, 2.1%, 3.3%, and 4.0%, respectively, of the entire platelet pool. To examine whether the iMK-derived platelets were incorporated into ex vivo thrombus formation, FITC-anti human CD41 antibody-labeled blood sample from iMK-infused thrombocytopenic NOG mouse was perfused on type I collagen-coated chip under flow condition (Total Thrombus-formation Analyzing System). After perfusion for 10 min, we observed that cells expressing human CD41 were incorporated into thrombi on collagen surface. These findings demonstrate that p45NF-E2 is a key determinant factor for megakaryopoiesis and thrombopoiesis, and is sufficient to lead to platelet formation from a number of non-hematopoietic adult tissues. Generation of iMKs from fibroblasts could have important implications for studying the mechanisms of MK differentiation and platelet production. Disclosures: No relevant conflicts of interest to declare.

RNA-Mediated Reprogramming of Primary Adult Human Dermal Fibroblasts into c-kit+ Cardiac Progenitor Cells

2015 ◽  
Vol 24 (22) ◽  
pp. 2622-2633 ◽  
Author(s):  
Elizabeth D. Pratico ◽  
Bryan J. Feger ◽  
Michael J. Watson ◽  
Bruce A. Sullenger ◽  
Dawn E. Bowles ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Dermal Fibroblasts ◽  
Cardiac Progenitor Cells ◽  
Human Dermal Fibroblasts ◽  
Adult Human

scholarly journals Adult human dermal fibroblasts exposed to nanosecond electrical pulses exhibit genetic biomarkers of mechanical stress

2017 ◽  
Vol 9 ◽  
pp. 302-309 ◽  
Author(s):  
Caleb C. Roth ◽  
Randolph D. Glickman ◽  
Stacey L. Martens ◽  
Ibtissam Echchgadda ◽  
Hope T. Beier ◽  
...  
Keyword(s):  
Mechanical Stress ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
Adult Human ◽  
Genetic Biomarkers ◽  
Electrical Pulses
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