Growth Factor- and Adhesion Protein-Like Components of Fetal Calf Serum Can Significantly Enhance the Intracellular Delivery of Peptide Nucleic Acids

2011 ◽  
Vol 21 (4) ◽  
pp. 285-291 ◽  
Author(s):  
Johannes Oehlke ◽  
Angelika Ehrlich ◽  
Eberhard Krause ◽  
Stephan Pritz ◽  
Burkhard Wiesner ◽  
...  
Keyword(s):  
Growth Factor ◽  
Nucleic Acids ◽  
Fetal Calf Serum ◽  
Peptide Nucleic Acids ◽  
Calf Serum ◽  
Intracellular Delivery ◽  
Adhesion Protein

scholarly journals Phospholipid Conjugate for Intracellular Delivery of Peptide Nucleic Acids

2009 ◽  
Vol 20 (9) ◽  
pp. 1729-1736 ◽  
Author(s):  
Gang Shen ◽  
Huafeng Fang ◽  
Yinyin Song ◽  
Agata A. Bielska ◽  
Zhenghui Wang ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Peptide Nucleic Acids ◽  
Intracellular Delivery

The Hyperresponsiveness of Cells Expressing Truncated Erythropoietin Receptors Is Contingent on Insulin-Like Growth Factor-1 in Fetal Calf Serum

Blood ◽  
1998 ◽  
Vol 92 (2) ◽  
pp. 425-433 ◽  
Author(s):  
Jacqueline E. Damen ◽  
Jana Krosl ◽  
Donna Morrison ◽  
Steven Pelech ◽  
Gerald Krystal
Keyword(s):  
Cell Death ◽  
Growth Factor ◽  
Neutralizing Antibodies ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Negative Regulator ◽  
Combined Effects ◽  
Insulin Like Growth Factor ◽  
Wild Type

Abstract We demonstrate herein that the well documented hyperresponsiveness to erythropoietin (Epo) of Ba/F3 cells expressing C-terminal truncated erythropoietin receptors (EpoRs) is contingent on these cells being in fetal calf serum (FCS). In the absence of FCS, their Epo-induced proliferation is far poorer than Ba/F3 cells expressing wild-type (WT) EpoRs. This hyporesponsiveness in the absence of serum is also seen in DA-3 cells expressing these truncated EpoRs. In fact, long-term proliferation studies performed in the absence of serum show that even at saturating concentrations of Epo, Ba/F3 cells expressing these truncated receptors die via apoptosis, while cells bearing WT EpoRs do not, and this programmed cell death correlates with an inability of Epo-stimulated Ba/F3 cells expressing truncated EpoRs to induce the tyrosine phosphorylation of MAPK and the activation of p70S6K. Using neutralizing antibodies to insulin-like growth factor (IGF)-1, we show that a major non-Epo factor in FCS that contributes to the hyperresponsive phenotype of Ba/F3 cells expressing truncated EpoRs is IGF-1. Our results suggest that the Epo-hypersensitivity of truncated EpoR expressing Ba/F3 cells is due to the combined effects of these EpoRs not possessing a binding site for the negative regulator, SHP-1, and the triggering of proliferation-inducing/apoptosis-inhibiting cascades, lost through EpoR truncation, by IGF-1.

scholarly journals A universal discoidal nanoplatform for the intracellular delivery of PNAs

Nanoscale ◽  
2019 ◽  
Vol 11 (26) ◽  
pp. 12517-12529 ◽  
Author(s):  
Armin Tahmasbi Rad ◽  
Shipra Malik ◽  
Lin Yang ◽  
Tripat Kaur Oberoi-Khanuja ◽  
Mu-Ping Nieh ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Gene Editing ◽  
Peptide Nucleic Acids ◽  
Intracellular Delivery

Peptide nucleic acids (PNAs) have gained considerable attention due to their remarkable potential in gene editing and targeting-based strategies.

The Hyperresponsiveness of Cells Expressing Truncated Erythropoietin Receptors Is Contingent on Insulin-Like Growth Factor-1 in Fetal Calf Serum

Blood ◽  
1998 ◽  
Vol 92 (2) ◽  
pp. 425-433
Author(s):  
Jacqueline E. Damen ◽  
Jana Krosl ◽  
Donna Morrison ◽  
Steven Pelech ◽  
Gerald Krystal
Keyword(s):  
Cell Death ◽  
Growth Factor ◽  
Neutralizing Antibodies ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Negative Regulator ◽  
Combined Effects ◽  
Insulin Like Growth Factor ◽  
Wild Type

We demonstrate herein that the well documented hyperresponsiveness to erythropoietin (Epo) of Ba/F3 cells expressing C-terminal truncated erythropoietin receptors (EpoRs) is contingent on these cells being in fetal calf serum (FCS). In the absence of FCS, their Epo-induced proliferation is far poorer than Ba/F3 cells expressing wild-type (WT) EpoRs. This hyporesponsiveness in the absence of serum is also seen in DA-3 cells expressing these truncated EpoRs. In fact, long-term proliferation studies performed in the absence of serum show that even at saturating concentrations of Epo, Ba/F3 cells expressing these truncated receptors die via apoptosis, while cells bearing WT EpoRs do not, and this programmed cell death correlates with an inability of Epo-stimulated Ba/F3 cells expressing truncated EpoRs to induce the tyrosine phosphorylation of MAPK and the activation of p70S6K. Using neutralizing antibodies to insulin-like growth factor (IGF)-1, we show that a major non-Epo factor in FCS that contributes to the hyperresponsive phenotype of Ba/F3 cells expressing truncated EpoRs is IGF-1. Our results suggest that the Epo-hypersensitivity of truncated EpoR expressing Ba/F3 cells is due to the combined effects of these EpoRs not possessing a binding site for the negative regulator, SHP-1, and the triggering of proliferation-inducing/apoptosis-inhibiting cascades, lost through EpoR truncation, by IGF-1.

213 IN VITRO DEVELOPMENT OF BOVINE EMBRYOS IN THE PRESENCE OF GROWTH HORMONE, INSULIN-LIKE GROWTH FACTOR-1, AND INSULIN IN OOCYTE MATURATION AND EMBRYO CULTURE MEDIA

2008 ◽  
Vol 20 (1) ◽  
pp. 186 ◽  
Author(s):  
C. B. Ponchirolli-Schneider ◽  
C. P. Freitas ◽  
F. C. Landim-Alvarenga
Keyword(s):  
Growth Hormone ◽  
Growth Factor ◽  
Production Rate ◽  
Dna Fragmentation ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Insulin Like Growth Factor ◽  
The Third ◽  
Fragmentation Rate

The addition of hormones and growth factors to bovine IVM and IVC media has been reported to affect early embryonic development by enhancing the blastocyst formation rate and quality of embryos produced. The purpose of this study was to investigate the influence of adding growth hormone (GH), insulin-like growth factor-1 (IGF-1), and insulin to IVM and IVC media. Blastocyst production rate and blastocyst quality, as verified by the number of cells with DNA fragmentation, were evaluated. Ovaries from an abattoir were transported to the laboratory and COC were selected and cultured in IVM medium 199 (Earle's salts, Sigma, St. Louis, MO, USA), 10% fetal calf serum (Sigma), 50 µg mL–1 of sodium pyruvate, 1 µg mL–1 of estradiol (Sigma), 50 µg mL–1 of hCG (Profasi hp�, 5000 IU, Serono Inc., Rockland, MA, USA), 5 µg mL–1 of FSH (Folltropin�, Vetrepharm, Ontario, Canada), and 75 µg mL–1 of gentamicin sulfate for 24 h. After IVF (18 h), zygotes were partially denuded and transferred to IVC medium HTF (HTF�, Irvine Scientific, Santa Ana, CA, USA) and BME (BME�, Sigma), in a 1:1 proportion (HTF:BME), 0.6% BSA (Sigma), 0.01% myoinositol (Sigma), and 75 µg mL–1 of gentamicin sulfate, at 38.5�C, in a humidified atmosphere of 5% CO2 in air, supplemented with 10% fetal calf serum at Day 3 of culture. Three different experiments were performed. The first and second experiments were analyzed using the chi-square test (P < 0.05). The third experiment was analyzed with the general linear model of SAS� (SAS Institute Inc., Cary, NC, USA) and the Tukey test (P < 0.1). In the first experiment, oocytes were cultured in IVM medium supplemented with GH (10 ng mL–1), IGF-1 (100 ng mL–1), insulin (1 µg mL–1), or all 3 combined. In the second experiment, IVC medium was supplemented with GH, IGF-1, insulin, or all 3 combined (same concentrations as above). In the third experiment, the quality of the embryos produced in the first 2 experiments was determined by the percentage of cells with DNA fragmentation. After 96 h of culture, embryos were stained with orange acridin (100 µg mL–1) and propidium iodide (100 µg mL–1) and slides were evaluated by fluorescence microscopy (450 to 490 nm). Rates of blastocyst production (blastocysts/oocytes) in the first experiment (29, 28, 28, 26, and 28% for control, GH, IGF-1, insulin, or all 3 combined, respectively) and in the second experiment (35, 35, 36, 35, and 31%) were not statistically different among the groups. In the third experiment, the addition of GH, IGF-1, or insulin to IVM medium did not affect the DNA fragmentation rate (11, 5, 2, 12, and 12%). However, the addition of insulin to IVC medium led to a higher DNA fragmentation rate (24%), when compared with the other groups (11, 10, 6, and 8% for control, GH, IGF-1, and all 3 combined). The addition of GH or IGF-1 to bovine IVM and IVC media did not affect the blastocyst production rate or the quality of embryos produced. The quality of embryos cultured in the presence of insulin was negatively affected.

A factor present in fetal calf serum enhances oncogene-induced transformation of rodent fibroblasts

1987 ◽  
Vol 7 (10) ◽  
pp. 3380-3385
Author(s):  
W L Hsiao ◽  
C A Lopez ◽  
T Wu ◽  
I B Weinstein
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Time Course ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Fold Increase ◽  
Fibroblast Cell ◽  
Focus Formation ◽  
Heat Stable

Our previous studies indicated that addition of the tumor promoters 12-O-tetradecanoylphorbol-13-acetate (TPA) or teleocidin to Dulbecco modified Eagle medium supplemented with calf serum enhanced T24-induced focus formation in both the murine C3H 10T1/2 and rat 6 embryo fibroblast cell lines. In the present studies we have found that fetal calf serum (FCS) is more potent than 12-O-tetradecanoylphorbol-13-acetate in enhancing T24-induced focus formation, in terms of the number and size of the foci, in both C3H 10T1/2 and rat 6 cells. Time course studies indicate that FCS can exert this enhancing effect when it is added several days after the transfection with T24 DNA. In rat 6 cells, an 11-fold increase in T24-induced focus formation occurred when the transfected cultures were maintained for only 1 day in 5% FCS, starting 4 days after the transfection. Several known growth factors, including epidermal growth factor, transforming growth factors alpha and beta, insulin, and platelet-derived growth factor, did not enhance T24-induced transformation in these cell systems. Fractionation studies indicate that the factor present in FCS has a molecular weight of about 1,300, is not lipid soluble, and is acid, base, and heat stable. These findings suggest that a factor(s) normally present in serum may enhance the emergence of tumor cells in vivo, by acting in concert with an activated oncogene, during the multistage carcinogenic process.

scholarly journals Role of Cell-Penetrating Peptides in Intracellular Delivery of Peptide Nucleic Acids Targeting Hepadnaviral Replication

2017 ◽  
Vol 9 ◽  
pp. 162-169 ◽  
Author(s):  
Bénédicte Ndeboko ◽  
Narayan Ramamurthy ◽  
Guy Joseph Lemamy ◽  
Catherine Jamard ◽  
Peter E. Nielsen ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Peptide Nucleic Acids ◽  
Intracellular Delivery ◽  
Cell Penetrating Peptides ◽  
Cell Penetrating

scholarly journals A factor present in fetal calf serum enhances oncogene-induced transformation of rodent fibroblasts.

1987 ◽  
Vol 7 (10) ◽  
pp. 3380-3385 ◽  
Author(s):  
W L Hsiao ◽  
C A Lopez ◽  
T Wu ◽  
I B Weinstein
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Time Course ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Fold Increase ◽  
Fibroblast Cell ◽  
Focus Formation ◽  
Heat Stable

Our previous studies indicated that addition of the tumor promoters 12-O-tetradecanoylphorbol-13-acetate (TPA) or teleocidin to Dulbecco modified Eagle medium supplemented with calf serum enhanced T24-induced focus formation in both the murine C3H 10T1/2 and rat 6 embryo fibroblast cell lines. In the present studies we have found that fetal calf serum (FCS) is more potent than 12-O-tetradecanoylphorbol-13-acetate in enhancing T24-induced focus formation, in terms of the number and size of the foci, in both C3H 10T1/2 and rat 6 cells. Time course studies indicate that FCS can exert this enhancing effect when it is added several days after the transfection with T24 DNA. In rat 6 cells, an 11-fold increase in T24-induced focus formation occurred when the transfected cultures were maintained for only 1 day in 5% FCS, starting 4 days after the transfection. Several known growth factors, including epidermal growth factor, transforming growth factors alpha and beta, insulin, and platelet-derived growth factor, did not enhance T24-induced transformation in these cell systems. Fractionation studies indicate that the factor present in FCS has a molecular weight of about 1,300, is not lipid soluble, and is acid, base, and heat stable. These findings suggest that a factor(s) normally present in serum may enhance the emergence of tumor cells in vivo, by acting in concert with an activated oncogene, during the multistage carcinogenic process.

Sign in / Sign up

Export Citation Format

Share Document