Immortalization of epithelial cells

1996 ◽  
Vol 270 (1) ◽  
pp. C1-C11 ◽  
Author(s):  
U. Hopfer ◽  
J. W. Jacobberger ◽  
D. C. Gruenert ◽  
R. L. Eckert ◽  
P. S. Jat ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Cell Lines ◽  
Cell Biology ◽  
Transgenic Animals ◽  
Early Passage ◽  
Tight Junction Formation ◽  
Genetic Abnormalities ◽  
Junction Formation ◽  
Polar Orientation ◽  
Passage Cells

The methodologies for isolating cell lines have become very powerful, particularly in terms of retaining differentiated features of the parent cells. Cell lines can be developed from primary or early passage cells as well as from transgenic animals that carry an immortalizing gene. Cell lines from epithelia have been selected for their polar orientation, tight junction formation, and expression of differentiated markers or functions. These cell lines provide useful models for studying cell biology of specific tissues, tumorigenicity, genetic abnormalities, or to help screen for effective methods of gene therapy.

scholarly journals 423. The Combination of p53 Dendritic Cell Vaccine and Ad-p53 Gene Therapy Against p53 Overexpressing and p53 Deleted Tumor Cell Lines

2015 ◽  
Vol 23 ◽  
pp. S167
Author(s):  
Hiroki Saito ◽  
Koichi Kitagawa ◽  
Risa Yamasaki ◽  
Nami Katai ◽  
Naoya Morishita ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Dendritic Cell ◽  
Tumor Cell ◽  
Cell Lines ◽  
P53 Gene ◽  
Dendritic Cell Vaccine ◽  
Cell Vaccine ◽  
Tumor Cell Lines

scholarly journals Conditionally immortalised leukaemia initiating cells co-expressing Hoxa9/Meis1 demonstrate microenvironmental adaptation properties ex vivo while maintaining myelomonocytic memory

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Maike Stahlhut ◽  
Teng Cheong Ha ◽  
Ekaterina Takmakova ◽  
Michael A. Morgan ◽  
Adrian Schwarzer ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Fate ◽  
Cell Lines ◽  
Cell Biology ◽  
Ex Vivo ◽  
Haematopoietic Stem Cell ◽  
Transcriptional Dysregulation ◽  
Conditional Gene Expression ◽  
Exponential Expansion ◽  
Drug Dependent

AbstractRegulation of haematopoietic stem cell fate through conditional gene expression could improve understanding of healthy haematopoietic and leukaemia initiating cell (LIC) biology. We established conditionally immortalised myeloid progenitor cell lines co-expressing constitutive Hoxa9.EGFP and inducible Meis1.dTomato (H9M-ciMP) to study growth behaviour, immunophenotype and morphology under different cytokine/microenvironmental conditions ex vivo upon doxycycline (DOX) induction or removal. The vector design and drug-dependent selection approach identified new retroviral insertion (RVI) sites that potentially collaborate with Meis1/Hoxa9 and define H9M-ciMP fate. For most cell lines, myelomonocytic conditions supported reversible H9M-ciMP differentiation into neutrophils and macrophages with DOX-dependent modulation of Hoxa9/Meis1 and CD11b/Gr-1 expression. Here, up-regulation of Meis1/Hoxa9 promoted reconstitution of exponential expansion of immature H9M-ciMPs after DOX reapplication. Stem cell maintaining conditions supported selective H9M-ciMP exponential growth. H9M-ciMPs that had Ninj2 RVI and were cultured under myelomonocytic or stem cell maintaining conditions revealed the development of DOX-dependent acute myeloid leukaemia in a murine transplantation model. Transcriptional dysregulation of Ninj2 and distal genes surrounding RVI (Rad52, Kdm5a) was detected. All studied H9M-ciMPs demonstrated adaptation to T-lymphoid microenvironmental conditions while maintaining immature myelomonocytic features. Thus, the established system is relevant to leukaemia and stem cell biology.

scholarly journals Participation of plasma membrane proteins in the formation of tight junction by cultured epithelial cells

1983 ◽  
Vol 96 (3) ◽  
pp. 693-702 ◽  
Author(s):  
EB Griepp ◽  
WJ Dolan ◽  
ES Robbins ◽  
DD Sabatini
Keyword(s):  
Protein Synthesis ◽  
Steady State ◽  
Tight Junction ◽  
Messenger Rna ◽  
Freeze Fracture ◽  
Epithelial Cell Line ◽  
Experimental Conditions ◽  
Tight Junction Formation ◽  
Junction Formation ◽  
Spinner Culture

Measurements of the transepithelial electrical resistance correlated with freeze-fracture observations have been used to study the process of tight junction formation under various experimental conditions in monolayers of the canine kidney epithelial cell line MDCK. Cells derived from previously confluent cultures and plated immediately after trypsin- EDTA dissociation develop a resistance that reaches its maximum value of several hundred ohms-cm(2) after approximately 24 h and falls to a steady-state value of 80-150 ohms- cm(2) by 48 h. The rise in resistance and the development of tight junctions can be completely and reversibly prevented by the addition of 10 μg/ml cycloheximide at the time of plating, but not when this inhibitor is added more than 10 h after planting. Thus tight junction formation consists of separable synthetic and assembly phases. These two phases can also be dissociated and the requirement for protein synthesis after plating eliminated if, following trypsinization, the cells are maintained in spinner culture for 24 h before plating. The requirement for protein synthesis is restored, however, if cells maintained in spinner culture are treated with trypsin before plating. Actinomycin D prevents development of resistance only in monolayers formed from cells derived from sparse rather than confluent cultures, but new mRNA synthesis is not required if cells obtained from sparse cultures are maintained for 24 h in spinner culture before plating. Once a steady-state resistance has been reached, its maintenance does not require either mRNA or protein synthesis; in fact, inhibition of protein synthesis causes a rise in the resistance over a 30-h period. Following treatments that disrupt the junctions in steady- state monolayers recovery of resistance also does not require protein synthesis. These observations suggest that proteins are involved in tight junction formation. Such proteins, which do not turn over rapidly under steady-state conditions, are destroyed by trypsinization and can be resynthesized in the absence of stable cell-cell or cell-substratum contact. Messenger RNA coding for proteins involved in tight junction formation is stable except when cells are sparsely plated, and can also be synthesized without intercellular contacts or cell-substratum attachment.

scholarly journals Safety and Efficacy of Suicide Gene Therapy with Adenosine Deaminase 5-Fluorocytosine Silmutaneously in in Vitro Cultures of Melanoma and Retinal Cell Lines

2014 ◽  
Vol 5 (5) ◽  
pp. 368-381 ◽  
Author(s):  
Antonios Sakkas ◽  
Paul Zarogoulidis ◽  
Kalliopi Domvri ◽  
Wolfgang Hohenforst-Schmidt ◽  
Dimitris Bougiouklis ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Adenosine Deaminase ◽  
Cell Lines ◽  
Suicide Gene ◽  
In Vitro Cultures ◽  
Suicide Gene Therapy ◽  
Retinal Cell ◽  
Safety And Efficacy

scholarly journals Biological cell-based screening for scientific membranal and cytoplasmatic markers using dielectric spectroscopy

2008 ◽  
Vol 2 (2) ◽  
pp. 111-121
Author(s):  
Ragini Raj Singh ◽  
◽  
Amit Ron ◽  
Nick Fishelson ◽  
Irena Shur ◽  
...  
Keyword(s):  
Cell Lines ◽  
Dielectric Spectroscopy ◽  
Cell Biology ◽  
Screening Tool ◽  
Excitation Frequency ◽  
Cell Physiology ◽  
Biological Cell ◽  
Non Invasive ◽  
Native Cell ◽  
Biology Research

Dielectric spectroscopy (DS) of living biological cells is based on the analysis of cells suspended in a physiological medium. It provides knowledge of the polarization-relaxation response of the cells to external electric field as function of the excitation frequency. This response is strongly affected by both structural and molecular properties of the cells and, therefore, can reveal rare insights into cell physiology and behaviour. This study demonstrates the mapping potential of DS after cytoplasmic and membranal markers for cell-based screening analysis. The effect of membrane permittivity and cytoplasm conductivity was examined using tagged MBA and MDCK cell lines respectively. The comparison of the dielectric spectra of tagged and native cell lines reveals clear differences between the cells. In addition, the differences in the matching dielectric properties of the cells were discovered. Those findings support the high distinction resolution and sensitivity of DS after fine molecular and cellular changes, and hence, highlight the high potential of DS as non invasive screening tool in cell biology research.

scholarly journals Design of time-delayed safety switches for CRISPR gene therapy

2021 ◽  
Author(s):  
Dashan Sun
Keyword(s):  
Gene Therapy ◽  
Immune Response ◽  
Time Delay ◽  
Cell Lines ◽  
Gene Editing ◽  
Drug Accumulation ◽  
Work Time ◽  
Crispr System ◽  
Efficient Gene ◽  
Target Effect

CRISPR system is a powerful gene editing tool which has already been reported to address a variety of gene relevant diseases in different cell lines. However, off-target effect and immune response caused by Cas9 remain two fundamental problems. In our work, time-delayed safety switches are designed based on either artificial ultrasensitivity transmission module or intrinsic time delay in biomolecular activities. By addressing gene therapy efficiency, off-target effect, immune response and drug accumulation, we hope our safety switches may offer inspiration in realizing safe and efficient gene therapy in humans.

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