scholarly journals Developing a novel serum-free cell culture model of skeletal muscle differentiation by systematically studying the role of different growth factors in myotube formation

2009 ◽  
Vol 45 (7) ◽  
pp. 378-387 ◽  
Author(s):  
Mainak Das ◽  
John W. Rumsey ◽  
Neelima Bhargava ◽  
Cassie Gregory ◽  
Lisa Riedel ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Cell Culture ◽  
Growth Factors ◽  
Free Cell ◽  
Skeletal Muscle Differentiation ◽  
Cell Culture Model ◽  
Serum Free ◽  
Myotube Formation ◽  
Culture Model

A Cell Culture Model for Studying the Role of Neuron-Glia Interactions in Ischemia

10.3791/61388 ◽  
2020 ◽  
Author(s):  
Genilso Gava-Junior ◽  
Cláudio Roque ◽  
Julieta Mendes-Oliveira ◽  
Ana C Bernardino ◽  
Inês Serrenho ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Culture Model ◽  
Culture Model ◽  
A Cell

Development of Unique Growth Factors for Serum-Free Cell Culture using Fusion Partner Technology

1997 ◽  
pp. 737-741
Author(s):  
G. L. Francis ◽  
C. Goddard ◽  
P. E. Walton ◽  
Z. Upton ◽  
F. J. Ballard
Keyword(s):  
Cell Culture ◽  
Growth Factors ◽  
Free Cell ◽  
Fusion Partner ◽  
Serum Free

scholarly journals S143 Premature Ageing And Skeletal Muscle Dysfunction In Copd Patients: Development Of A Cell Culture Model

Thorax ◽  
2014 ◽  
Vol 69 (Suppl 2) ◽  
pp. A76-A76
Author(s):  
R. Lahkdar ◽  
G. Choudhury ◽  
J. McLeish ◽  
E. Drost ◽  
L. McGlynn ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Cell Culture ◽  
Muscle Dysfunction ◽  
Cell Culture Model ◽  
Premature Ageing ◽  
Skeletal Muscle Dysfunction ◽  
Copd Patients ◽  
Culture Model ◽  
A Cell

scholarly journals Adipose-derived stem cells and keratinocytes in a chronic wound cell culture model: the role of hydroxyectoine

2013 ◽  
Vol 12 (4) ◽  
pp. 387-396 ◽  
Author(s):  
Oliver C Thamm ◽  
Panagiotis Theodorou ◽  
Ewa Stuermer ◽  
Max J Zinser ◽  
Edmund A Neugebauer ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Chronic Wound ◽  
Adipose Derived Stem Cells ◽  
Cell Culture Model ◽  
Culture Model

Insulin-like Growth Factors (IGF) I and II Utilize Different Calcium Signaling Pathways in a Primary Human Parathyroid Cell Culture Model

2006 ◽  
Vol 30 (3) ◽  
pp. 333-345 ◽  
Author(s):  
C. K. M. Wong ◽  
T. Lai ◽  
J. M. P. Holly ◽  
M. H. Wheeler ◽  
C. E. H. Stewart ◽  
...  
Keyword(s):  
Cell Culture ◽  
Growth Factors ◽  
Calcium Signaling ◽  
Signaling Pathways ◽  
Parathyroid Cell ◽  
Cell Culture Model ◽  
Culture Model ◽  
Igf I ◽  
Insulin Like Growth Factors

scholarly journals Functional Analysis of Hepatitis C Virus (HCV) Envelope Protein E1 Using a trans-Complementation System Reveals a Dual Role of a Putative Fusion Peptide of E1 in both HCV Entry and Morphogenesis

2017 ◽  
Vol 91 (7) ◽  
Author(s):  
Yimin Tong ◽  
Xiaojing Chi ◽  
Wei Yang ◽  
Jin Zhong
Keyword(s):  
Cell Culture ◽  
Life Cycle ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Ectopic Expression ◽  
Fusion Peptide ◽  
Hcv Infection ◽  
Cell Culture Model ◽  
Culture Model

ABSTRACT Hepatitis C virus (HCV) is an enveloped RNA virus belonging to the Flaviviridae family. It infects mainly human hepatocytes and causes chronic liver diseases, including cirrhosis and cancer. HCV encodes two envelope proteins, E1 and E2, that form a heterodimer and mediate virus entry. While E2 has been extensively studied, less has been done so for E1, and its role in the HCV life cycle still needs to be elucidated. Here we developed a new cell culture model for HCV infection based on the trans-complementation of E1. Virus production of the HCV genome lacking the E1-encoding sequence can be efficiently rescued by the ectopic expression of E1 in trans. The resulting virus, designated HCVΔE1, can propagate in packaging cells expressing E1 but results in only single-cycle infection in naive cells. By using the HCVΔE1 system, we explored the role of a putative fusion peptide (FP) of E1 in HCV infection. Interestingly, we found that the FP not only contributes to HCV entry, as previously reported, but also may be involved in virus morphogenesis. Finally, we identified amino acid residues in FP that are critical for biological functions of E1. In summary, our work not only provides a new cell culture model for studying HCV but also provides some insights into understanding the role of E1 in the HCV life cycle. IMPORTANCE Hepatitis C virus (HCV), an enveloped RNA virus, encodes two envelope proteins, E1 and E2, that form a heterodimeric complex to mediate virus entry. Compared to E2, the biological functions of E1 in the virus life cycle are not adequately investigated. Here we developed a new cell culture model for single-cycle HCV infection based on the trans-complementation of E1. The HCV genome lacking the E1-encoding sequence can be efficiently rescued for virus production by the ectopic expression of E1 in trans. This new model renders a unique system to dissect functional domains and motifs in E1. Using this system, we found that a putative fusion peptide in E1 is a multifunctional structural element contributing to both HCV entry and morphogenesis. Our work has provided a new cell culture model to study HCV and provides insights into understanding the biological roles of E1 in the HCV life cycle.

Influence of Prion Gene Downregulation by RNAi on Erythroid Differentiation in Vitro. First Insight.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1338-1338
Author(s):  
Marti N Panigaj ◽  
Hana Glierova ◽  
Karel Holada
Keyword(s):  
Cell Culture ◽  
Cell Survival ◽  
Cell Lines ◽  
Erythroid Differentiation ◽  
Cell Culture Model ◽  
Packaging Cell Line ◽  
Prion Gene ◽  
Qrt Pcr ◽  
Culture Model

Abstract Cellular prion protein (PrPc) has significant medical importance but its physiological role remains unclear. Some reports indicate that PrPc may play role in the cell survival and/or differentiation. Connection between prion pathogenesis and erythropoiesis was suggested by downregulation of alpha hemoglobin stabilizing protein (AHSP) during prion disease. In addition we recently demonstrated the reduction of erythroid cell and erythropoietin production in PrP-null mice in response to acute anemia (Zivny et al. BCMD 40(2008)302–7). Supporting data for possible involvement of PrPc in hematopoiesis come from the study showing that long-term hematopoietic stem cells of PrP-null bone marrow exhibited impaired self-renewal in serial transplantation of lethally irradiated mouse recipients. Pilot studies in cell culture model, represented by murine erythroleukemic (MEL) cells, indicated upregulation of PrPc expression during erythroid differentiation suggesting its importance in the process. In order to study the role of PrPc during the erythroid differentiation we created 3 stable transfected MEL lines infected with retroviruses coding the short interfering RNA in context of miRNA (1 nonsilencing control - LN and 2 RNAs - LP1 and LP2 targeting murine prion gene (Prnp)). Anti Prnp sequences (HP_285770 and HP_288208) were chosen from publicly available database (RNAi codex). As a vector we used MSCV/LTRmiR30-PIG (Open Biosystems) retroviral plasmid which was transfected to packaging cell line HEK293GP2 and medium containing the viruses was used to infect the MEL cells. Upon selection with 0.5 μg/mL of Puromycin we obtained ~95% of positive cells (estimated by eGFP expression). Downregulation of PrPc was verified by western blot and qRT-PCR. Significant inhibition of PrPc extending from ~70 to more than 90 % compared to LN - MEL cells was observable during the 6 day course of the differentiation (Fig.). We analyzed the expression of 3 genes by qRT-PCR: c-myb, AHSP and hemoglobin alpha (HBA). C-myb is required for early cellular expansion and its downregulation allows the terminal differentiation of cells. Interestingly on the start of differentiation we found ~70% upregulation of c-myb in lines with silenced PrPc. After entering of cells to differentiation, all lines downregulated the c-myb, but the decrease in c-myb expression was more pronounced in LP1 and LP2 cells (15% of initial expression after 24 hours) in comparison with LN cells (30%). Analysis of AHSP and HBA, showed similar upregulation in all cell lines. Since many reports suggest cytoprotective role of PrPc in prevention of apoptosis we analyzed potential influence of PrPc on MEL cell survival. Previous study showed that neither physiological upregulation nor enhanced expression by exogenous delivered Prnp cDNA altered percentage of apoptotic MEL cells during differentiation. We wondered if opposite approach- downregulation of PrPc will affect cell survival. Flow cytometry analysis upon cell staining with 7-AAD and Hoechst 33342 showed that inhibition of translation of PrPc during differentiation probably does not play role in sensitization of cells to apoptosis under physiological conditions. We hypothesized that if under normal conditions cell lines do not differ in level of apoptosis, then they could react differentially after exposure to external stress. We promoted the disturbing conditions by several different treatments: oxidative stress by presence of H2O2 (500 μM), Staurosporin (125 nM) and elevated incubation temperature (40°C). We also treated uninduced cells with increasing concentration of Cu2+ or exposed them to serum deprivation. Contrary to belief that absence of PrPc sensitizes the cells against damage, we found that inhibition of PrPc expression did not altered level of apoptosis/necrosis in MEL cells. Finally, we have introduced the new cell culture model for study of PrPc involvement in erythroid differentiation. Our initial observations suggest that direct cytoprotection is not mode of PrPc action in studied process, but it may play role in cell cycle which is a subject of undergoing research. Figure Figure

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