Isolation and characterization of a spontaneously immortalized bovine retinal pigmented epithelial cell line

Liggett, Thomas; Griffiths, Tony; Gaillard, Elizabeth R.

doi:10.1186/1471-2121-10-33

Cited by 11 publications

(9 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In our study, over 90% of canine RPE cells in passage 1 expressed the cytoplasmic RPE65 protein, as shown by both flow cytometry and immunocytochemistry. Furthermore, a decreasing expression of PRE65 and decreasing pigment production was seen in the following passages in longitudinal culture, which is in line with studies of human and bovine RPE cells (Alge et al 2003;Hunt and Davis 1990;Liggett et al 2009;Tamiya Fig. 6 Leucinostatin co-induced Hsp70 expression in dog primary RPE cells (passages 2-4).…”

Section: Discussionsupporting

confidence: 85%

Leucinostatin acts as a co-inducer for heat shock protein 70 in cultured canine retinal pigment epithelial cells

Lyu

Ludwig

Kooten

et al. 2020

Cell Stress and Chaperones

View full text Add to dashboard Cite

Dysregulation of retinal pigment epithelium (RPE) cells is the main cause of a variety of ocular diseases. Potentially heat shock proteins, by preventing molecular and cellular damage and modulating inflammatory disease, may exert a protective role in eye disease. In particular, the inducible form of heat shock protein 70 (Hsp70) is widely upregulated in inflamed tissues, and in vivo upregulation of Hsp70 expression by HSP co-inducing compounds has been shown to be a potential therapeutic strategy for inflammatory diseases. In order to gain further understanding of the potential protective effects of Hsp70 in RPE cells, we developed a method for isolation and culture of canine RPE cells. Identity of RPE cells was confirmed by detection of its specific marker, RPE65, in qPCR, flow cytometry, and immunocytochemistry analysis. The ability of RPE cells to express Hsp70 upon experimental induction of cell stress, by arsenite, was analyzed by flow cytometry. Finally, in search of a potential Hsp70 coinducer, we investigated whether the compound leucinostatin could enhance Hsp70 expression in stressed RPE cells. Canine RPE cells were isolated and cultured successfully. Purity of cells that strongly expressed RPE65 was over 90%. Arsenite-induced stress led to a time-and dose-dependent increase in Hsp70 expression in canine RPE cells in vitro. In addition, leucinostatin, which enhanced heat shock factor-1-induced transcription from the heat shock promoter in DNAJB1-luc-O23 reporter cell line, also enhanced Hsp70 expression in arsenite-stressed RPE cells, in a dose-dependent fashion. These findings demonstrate that leucinostatin can boost Hsp70 expression in canine RPE cells, most likely by activating heat shock factor-1, suggesting that leucinostatin might be applied as a new co-inducer for Hsp70 expression.

show abstract

Section: Discussionsupporting

confidence: 85%

Leucinostatin acts as a co-inducer for heat shock protein 70 in cultured canine retinal pigment epithelial cells

Lyu

Ludwig

Kooten

et al. 2020

Cell Stress and Chaperones

View full text Add to dashboard Cite

show abstract

“…Unfortunately, primary cells of RPE lose many of their in vivo state characteristics within the first few passages in culture. These changes include depigmentation and alteration in metabolic functions, gene expression and morphology (Alge et al, ; Liggett et al, ). Because of the disadvantages of short‐term cultures and the lack of sufficient donor eye tissue, immortalized cell lines would be readily available source of RPE cells for long‐term studies.…”

Section: Discussionmentioning

confidence: 99%

“…They lose many of their in vivo state characteristics within the first few passages. These changes include depigmentation and an alteration in metabolic function, gene expression, and morphology (Hunt and Davis, ; Rizzolo, ; Huotari et al, ; Alge et al, ; Liggett et al, ). They rarely survive more than eight to ten passages.…”

mentioning

confidence: 99%

Isolation, Characterization, and Establishment of Spontaneously Immortalized Cell Line HRPE‐2S With Stem Cell Properties

Najafabadi

Soheili

Samiei

et al. 2017

Journal Cellular Physiology

View full text Add to dashboard Cite

The retinal pigment epithelium is a monolayer of highly specialized pigmented cells located between the neural retina and the Bruch's membrane of the choroid. RPE cells play a crucial role in the maintenance and function of the underlying photoreceptors. This study introduces a spontaneously arising human retinal pigment epithelial cell line, HRPE-2S, which was isolated from primary RPE cell culture of 2 days old male donor. We characterized morphology and functional properties of the new cell line. The immortalized cell line was maintained in culture for more than 70 passages and 240 divisions. The average doubling time of the cells was approximately 22 h and got freezed at 26th passage. The cell line expressed RPE-specific markers RPE65 and cell junction protein ZO1 as an epithelial cell marker. It also expressed CHX10, PAX6, Nestin, SOX2 as stem and retinal progenitor cell markers. Ki67 as a marker of cell proliferation was expressed in all HRPE-2S cells. It represented typical epithelial cobblestone morphology and did not phenotypically change through several passages. Stem cell-like aggregations (neurospheres) were observed in SEM microscopy. The cells represented high mitotic index. They could be viable under hypoxic conditions and serum deprivation. According to functional studies, the cell line exhibited stem cell-like behaviors with particular emphasis on its self-renewal capacity. LDH isoenzymes expression pattern confirmed the same cellular source for both of the HRPE-2S cells and primary RPE cells. Characteristics of HRPE-2S cells promise it as an in vitro model for RPE stem cell-based researches. J. Cell. Physiol. 232: 2626-2640, 2017. © 2016 Wiley Periodicals, Inc.

show abstract

“…Immunocytochemical staining can provide a comprehensive characterization of the cell types of mammary glands (Li et al, ). Cytokeratins are proteins of keratin‐containing intermediate filaments found in the intracytoplasmic cytoskeleton of epithelial tissue most exclusively, while vimentin is usually used as a marker for the determination of cell types of from mesenchymal origin (Liggett et al, ). Both of these proteins are frequently coexpressed in many epithelial cells.…”

Section: Discussionmentioning

confidence: 99%

“…Although morphology and tumorigenicity characteristics never changed, the variation of chromosome numbers and cell proliferation was relevant to the stable maintenance of chromosomes of CMEC-H as an immortalized cell line.Immunocytochemical staining can provide a comprehensive characterization of the cell types of mammary glands (Li et al, 1999). Cytokeratins are proteins of keratin-containing intermediate filaments found in the intracytoplasmic cytoskeleton of epithelial tissue most exclusively, while vimentin is usually used as a marker for the determination of cell types of from mesenchymal origin (Liggett et al, 2009). Both of these proteins are frequently coexpressed in many epithelial cells.…”

Section: Discussionmentioning

confidence: 99%

Immortalized bovine mammary epithelial cells express stem cell markers and differentiate in vitro

Zheng

Gao

et al. 2016

Cell Biology International

View full text Add to dashboard Cite

The bovine mammary epithelial cell is a secretory cell, and its cell number and secretory activity determine milk production. In this study, we immortalized a bovine mammary epithelial cell line by SV40 large T antigen gene using a retrovirus based on Chinese Holstein primary mammary epithelial cells (CMEC) cultured in vitro. An immortalized bovine mammary epithelial cell line surpassed the 50-passage mark and was designated the CMEC-H. The immortalized mammary epithelial cells grew in close contact with each other and exhibited the typical cobblestone morphology characteristic with obvious boundaries. The telomerase expression of CMEC-H has consistently demonstrated the presence of telomerase activity as an immortalized cell line, but the cell line never induced tumor formation in nude mice. CMEC-H expressed epithelial (cytokeratins CK7, CK8, CK18, and CK19), mesenchymal (vimentin), and stem/progenitor (CD44 and p63) cell markers. The induced expression of milk proteins, αS1 -casein, β-casein, κ-casein, and butyrophilin, indicated that CMEC-H maintained the synthesis function of the mammary epithelial cells. The established immortalized bovine mammary epithelial cell line CMEC-H is capable of self-renewal and differentiation and can serve as a valuable reagent for studying the physiological mechanism of the mammary gland.

show abstract

Isolation and characterization of a spontaneously immortalized bovine retinal pigmented epithelial cell line

Cited by 11 publications

References 59 publications

Leucinostatin acts as a co-inducer for heat shock protein 70 in cultured canine retinal pigment epithelial cells

Leucinostatin acts as a co-inducer for heat shock protein 70 in cultured canine retinal pigment epithelial cells

Isolation, Characterization, and Establishment of Spontaneously Immortalized Cell Line HRPE‐2S With Stem Cell Properties

Immortalized bovine mammary epithelial cells express stem cell markers and differentiate in vitro

Contact Info

Product

Resources

About