Immortalization by large T-antigen of the adult epididymal duct epithelium

Kirchhoff, Christiane; Araki, Yoshihiko; Huhtaniemi, Ilpo; Matusik, Robert J.; Osterhoff, Caroline; Poutanen, Matti; Samalecos, Annemarie; Sipilä, Petra; Suzuki, Kichiya; Orgebin‐Crist, Marie‐Claire

doi:10.1016/j.mce.2003.10.073

Cited by 29 publications

(20 citation statements)

References 88 publications

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“…Thus, it seems plausible that similar interactions occur in other reproductive tissues, including the epididymis; however, this remains to be examined. The characterization of recently established epididymal cell lines should facilitate further investigation into the interactions between androgen and growth factor signaling pathways in the epididymis (Kirchhoff et al 2004, Tabuchi et al 2005.…”

Section: Growth Factor Systems In the Epididymismentioning

confidence: 99%

Region-specific expression of androgen and growth factor pathway genes in the rat epididymis and the effects of dual 5α-reductase inhibition

Henderson¹,

Cooke²,

Robaire³

2006

Journal of Endocrinology

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Dihydrotestosterone (DHT) is the primary androgen acting in the epididymis, the site of sperm maturation. Previously, we showed that the treatment of male rats with PNU157706, an inhibitor that acts on both isoforms of 5a-reductase to prevent DHT formation, has effects on the expression of genes implicated in processes that create the optimal luminal microenvironment required for sperm maturation, and on sperm maturation itself. However, signaling pathways involved in regulating or mediating DHT actions in the epididymis remain largely unknown. The goals of this study were to determine the expression profiles of potential signaling systems in the epididymis and assess their DHTdependence using two different dual 5a-reductase inhibitors. Rats were untreated or gavaged with vehicle, 10 mg/kg per day PNU157706 or 32 mg/kg per day FK143 for 28 days and epididymal gene expression was analyzed. Gene array analysis revealed analogous effects of FK143 on overall epididymal gene expression when compared with previous PNU157706 studies. Quantitative RT-PCR analysis of the expression of the 5a-reductase isozymes, androgen receptor, and members of the IGF, FGF, TGF, and VEGF families revealed novel region-specific expression profiles in the epididymis that were differentially affected by 5a-reductase inhibition; the two inhibitors had parallel effects. Specifically, in proximal regions, 5a-reductase 1, androgen receptor, and TGF-b1 expression increased after treatment, while in distal regions expression of IGF-I, IGFBP-5, IGFBP-6, and FGF-10 decreased. These results provide insight into epididymal signaling mechanisms and indicate potential candidates acting either upstream or downstream of DHT to regulate and/or mediate its actions in the epididymis.

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Section: Growth Factor Systems In the Epididymismentioning

confidence: 99%

Region-specific expression of androgen and growth factor pathway genes in the rat epididymis and the effects of dual 5α-reductase inhibition

Henderson¹,

Cooke²,

Robaire³

2006

Journal of Endocrinology

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“…The identification of SV40LT for cell line construction has been regarded a research highlight in the tissue engineering field since Gould first proposed the concept of "cell immortalization" [5,17,18].…”

Section: Discussionmentioning

confidence: 99%

“…Transfection of cells with SV40LT alone can lead to cell immortalization and this method has already been widely used to establish tissue cell lines [5]. However, immortalization efficiency using this approach is low [6] and the method has not been used to establish amniocyte lines with chromosome abnormalities.…”

Section: Introductionmentioning

confidence: 99%

An efficient method for immortalization of amniocytes with chromosome abnormalities for renewable application of rare disease sources

Weng

Shen

Chen

et al. 2019

Preprint

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Objective: We describe a new method of immortalizing amniocytes with chromosome abnormalities to generate renewable resources for studying rare diseases. Methods: Three methods were investigated by randomly dividing 180 cases of adherent amniocytes into groups A, B, and C. Group A cells were digested with 0.25% trypsin for 10, 20, 30, 60, 90, and 120 mins. Group B and C cells were digested with 0.25% trypsin for 3–5 minutes. Group A and B cells were then transfected with PT67 cell-produced SV40LT. Group C digested cells underwent liposome-mediated SV40LT-transfection. Results: The percentage of clones immortalized by SV40LT transfection in groups A, B, and C were 18.3, 5.0, and 16.7%, respectively, after G418 screening. Group A produced a higher percentage of immortalized clones than did groups B and C, but only significantly differed from group B. The karyotypes and chromosome 13, 18, 21, X, and Y fluorescence signals in transfected cells of all groups were identical to those of the primary cells after passaging for 10–15 generations. Conclusions: The trypsin-mediated SV40LT transfection used in group A can be applied for routine establishment of amniocyte lines to obtain a renewable resource for the study of rare diseases and as quality control materials for prenatal diagnosis.

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“…Immortalized cells or established cell lineages are cells that can grow in vitro indefinitely due to natural or induced transformation (e.g., embryonic stem cells or viral transformation) [34]. Immortalized cells have several advantages over primary cells, such as profitability, easy manipulation, unlimited supply of material, and lack of ethical concerns associated with the use of animal and human tissues [35]. Immortalized cells also provide a uniform and homogeneous population of cells [36].…”

Section: Stem Cell Culture Methodsmentioning

confidence: 99%

Traditional and Advanced Cell Cultures in Hematopoietic Stem Cell Studies

Ribeiro-Filho

Levy

Ruiz

et al. 2019

Cells

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Hematopoiesis is the main function of bone marrow. Human hematopoietic stem and progenitor cells reside in the bone marrow microenvironment, making it a hotspot for the development of hematopoietic diseases. Numerous alterations that correspond to disease progression have been identified in the bone marrow stem cell niche. Complex interactions between the bone marrow microenvironment and hematopoietic stem cells determine the balance between the proliferation, differentiation and homeostasis of the stem cell compartment. Changes in this tightly regulated network can provoke malignant transformation. However, our understanding of human hematopoiesis and the associated niche biology remains limited due to accessibility to human material and the limits of in vitro culture models. Traditional culture systems for human hematopoietic studies lack microenvironment niches, spatial marrow gradients, and dense cellularity, rendering them incapable of effectively translating marrow physiology ex vivo. This review will discuss the importance of 2D and 3D culture as a physiologically relevant system for understanding normal and abnormal hematopoiesis.

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Immortalization by large T-antigen of the adult epididymal duct epithelium

Cited by 29 publications

References 88 publications

Region-specific expression of androgen and growth factor pathway genes in the rat epididymis and the effects of dual 5α-reductase inhibition

Region-specific expression of androgen and growth factor pathway genes in the rat epididymis and the effects of dual 5α-reductase inhibition

An efficient method for immortalization of amniocytes with chromosome abnormalities for renewable application of rare disease sources

Traditional and Advanced Cell Cultures in Hematopoietic Stem Cell Studies

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