Testicular organoid generation by a novel in vitro three-layer gradient system

Alves-Lopes, João Pedro; Söder, Olle; Stukenborg, Jan-Bernd

doi:10.1016/j.biomaterials.2017.03.025

Cited by 128 publications

(136 citation statements)

References 35 publications

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“…Studies by Clever and collaborators, in which single intestinal stem cells were cultured in 3D, led to the demonstration that these cells could proliferate and differentiate into crypt–villus structures composed of four types of mature cells, thus reproducing the in vivo structure of the intestinal epithelium (Sato et al ., ). These structures were named organoids, or mini‐organs, and have since been developed using adult stem cells from the stomach, liver, brain, prostate, mammary gland, testis, endometrium, and fallopian tube, as well as other tissues (Barker et al ., ; Lukacs et al ., ; Hu et al ., ; Ewald, ; Huch et al ., ; Lancaster et al ., ; Kessler et al ., ; Alves‐Lopes et al ., ; Boretto et al ., ). Organoids can also be formed by 3D cell culture of embryonic stem cells, which can be differentiated into specific organ‐like structures by modifying the differentiation factors in the cell culture medium (Kim & Dressler, ).…”

Section: Epididymal Organoidsmentioning

confidence: 98%

Tissue regeneration and the epididymal stem cell

2019

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Background: In most pseudostratified epithelia, basal cells represent a multipotent adult stem cell population. These cells generally remain in a quiescent state, until they are stimulated to respond to tissue damage by initiating epithelial regeneration. In the epididymis, cell proliferation occurs at a relatively slow rate under normal physiological conditions. Epididymal basal cells have been shown to share common properties with multipotent adult stem cells. The development of organoids from stem cells represents a novel approach for understanding cellular differentiation and characterization of stem cells. Objective: To review the literature on tissue regeneration in the epididymis and demonstrate the presence of an epididymal stem cell population. Methods: PubMed database was searched for studies reporting on cell proliferation, regeneration, and stem cells in the epididymis. Three-dimensional cell culture of epididymal cells was used to determine whether these can develop into organoids in a similar fashion to stem cells from other tissues. Results: The epididymal epithelium can rapidly regenerate following orchidectomy or efferent duct ligation, in order to maintain epithelial integrity. Studies have isolated a highly purified fraction of rat epididymal basal cells and reported that these cells displayed properties similar to those of multipotent adult stem cells. In two-dimensional cell culture conditions, these cells differentiated into cells which expressed connexin 26, a marker of columnar cells, and cytokeratin 8. Furthermore, three-dimensional cell culture of epididymal cells resulted in the formation of organoids, a phenomenon associated with the proliferation and differentiation of stem cells in vitro. Conclusions: The rapid proliferation and tissue regeneration of the epididymal epithelium to preserve its integrity following tissue damage as well as the ability of cells to differentiate into organoids in vitro support the notion of a resident progenitor/stem cell population in the adult epididymis. Figure 7 (A) 3D cell culture of dispersed single epididymal cells. Bright-field images of epididymal spheres and organoids differentiated under 3D culture in Matrigel and a commercial cell culture medium (Prostacult, Stem cell Technologies, Vancouver, BC) supplemented with EGF, bFGF, and heparin. After 2 days of culture, two types of structures of <100 lm can be seen: 1. structures with irregular shape and; 2. spheres with a smooth outline. These structures then become more regular and can reach or exceed 100 lm in diameter after 4-6 days in culture. Scale bar, 100 lm. (B) Immunolocalization of Cx43 in a typical organoid following 11 days of culture. Cx43 is normally expressed in basal cells, and is shown to localize in the cells at the base of the organoid thereby indicating cell sorting and self-organization of the organoid. ANDROLOGY

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Section: Epididymal Organoidsmentioning

confidence: 98%

Tissue regeneration and the epididymal stem cell

2019

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“…In this approach, rat testis cells were suspended in Matrigel and placed between two cell-free layers of Matrigel, which after 7 days led to the formation of testicular organoids. Additionally, self-organization of testicular cells also led to eBTB formation and Sertoli cell epithelization; however, complete differentiation of germ cells was not observed [120]. These results collective suggest that maintaining the microenvironment of testicular cells even in the form of a 3D-culture system is crucial in achieving spermatogenesis ex vivo.…”

Section: Three-dimensional Culture Of Testis Cell Suspensionsmentioning

confidence: 87%

Spermatogonial Stem Cells for In Vitro Spermatogenesis and In Vivo Restoration of Fertility

Ibtisham

Honaramooz

2020

Cells

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Spermatogonial stem cells (SSCs) are the only adult stem cells capable of passing genes onto the next generation. SSCs also have the potential to provide important knowledge about stem cells in general and to offer critical in vitro and in vivo applications in assisted reproductive technologies. After century-long research, proof-of-principle culture systems have been introduced to support the in vitro differentiation of SSCs from rodent models into haploid male germ cells. Despite recent progress in organotypic testicular tissue culture and two-dimensional or three-dimensional cell culture systems, to achieve complete in vitro spermatogenesis (IVS) using non-rodent species remains challenging. Successful in vitro production of human haploid male germ cells will foster hopes of preserving the fertility potential of prepubertal cancer patients who frequently face infertility due to the gonadotoxic side-effects of cancer treatment. Moreover, the development of optimal systems for IVS would allow designing experiments that are otherwise difficult or impossible to be performed directly in vivo, such as genetic manipulation of germ cells or correction of genetic disorders. This review outlines the recent progress in the use of SSCs for IVS and potential in vivo applications for the restoration of fertility.Cells 2020, 9, 745 2 of 31 manipulation. Moreover, optimal conditions for the in vitro culture and propagation of SSCs are also needed to help boost the potential therapeutic application of SSCs in fertility preservation or restoration. Long-term culture of rodent SSCs is well demonstrated; however, relatively few in vitro culture conditions have been examined for primate SSCs [2]. Any potential in vivo application of cultured human SSCs, such as in restoration of fertility, requires extensive studies to ensure their safety and efficacy. The establishment of efficient culture conditions for human SSCs also necessitates the availability of proper animal models, for instance, xenotransplantation techniques to assess the quality and quantity of such cultured SSCs; these assays have so far been used sporadically for testing primate SSCs [4].Spermatogenesis is a complex process of germ cell proliferation and differentiation that requires extensive interactions among different cell types, hormones, growth factors, and various other signals, making it difficult to be replicated in vitro. There are several objectives for establishing an optimal and efficient culture system to recapitulate the process of germ cell development in vitro. This includes the study of basic requirements of male germ cell development, proliferation, differentiation, and production of haploid germ cells in a controlled in vitro environment. Additionally, such culture systems could be potentially used to produce haploid male germ cells from undifferentiated germ cells isolated from the testis of infertile adult patients and/or testicular biopsies collected from prepubertal cancer patients before undergoing gonadotoxic treatment. The establishment of ...

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“…While research with organoids mainly has focused on tissue engineering and regeneration, [148, 167] there is also a significant clinical need for biomimetic tumor models to bridge the technological gap between standard 2D cultures, 3D cultures such as spheroids, and in vivo models of cancer generated from established cell lines. However, compared to the large body of work using tumor spheroids, very few studies have attempted to engineer spatio-temporally organized organoid platforms to recapitulate complex tumor microenvironments.…”

Section: Organoid Modelsmentioning

confidence: 99%

Biomaterials‐Based Approaches to Tumor Spheroid and Organoid Modeling

Thakuri

Liu

Luker

et al. 2017

Adv Healthcare Materials

109

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Evolving understanding of structural and biological complexity of tumors has stimulated development of physiologically relevant tumor models for cancer research and drug discovery. A major motivation for developing new tumor models is to recreate the 3D environment of tumors and context-mediated functional regulation of cancer cells. Such models overcome many limitations of standard monolayer cancer cell cultures. Under defined culture conditions, cancer cells self-assemble into 3D constructs known as spheroids. Additionally, cancer cells may recapitulate steps in embryonic development to self-organize into 3D cultures known as organoids. Importantly, spheroids and organoids reproduce morphology and biologic properties of tumors, providing valuable new tools for research, drug discovery, and precision medicine in cancer. This Progress Report discusses uses of both natural and synthetic biomaterials to culture cancer cells as spheroids or organoids, specifically highlighting studies that demonstrate how these models recapitulate key properties of native tumors. The report concludes with the perspectives on the utility of these models and areas of need for future developments to more closely mimic pathologic events in tumors.

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Testicular organoid generation by a novel in vitro three-layer gradient system

Cited by 128 publications

References 35 publications

Tissue regeneration and the epididymal stem cell

Tissue regeneration and the epididymal stem cell

Spermatogonial Stem Cells for In Vitro Spermatogenesis and In Vivo Restoration of Fertility

Biomaterials‐Based Approaches to Tumor Spheroid and Organoid Modeling

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