The use of KnockOut serum replacement (KSR) in three dimensional rat testicular cells co-culture model: An improved male reproductive toxicity testing system

Zhang, Xiaofang; Wang, Lei; Zhang, Xiao Dong; Ren, Lijun; Shi, Wenjing; Tian, Yijun; Zhu, Jie; Zhang, Tianbao

doi:10.1016/j.fct.2017.05.001

Cited by 25 publications

(12 citation statements)

References 25 publications

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“…Later, Yokonishi et al (2013) used KSR supplementation in murine testis organoid system, which resulted in the formation of a compartmentalized testis organoid with tubular structures and partial spermatogenic differentiation [38]. Most testis organoid systems have used KSR as a media supplement to maintain spermatogonial proliferation in organoids [1,17], or to support spermatogenic differentiation from spermatogonia up to spermatocytes [49][50][51], spermatids [52], or even elongated spermatids [53]. Zhang et al (2014) also showed that de novo testis tubulogenesis and the formation of compartmentalized seminiferous tubule-like structures only occurred in KSR supplemented groups [49].…”

Section: Discussionmentioning

confidence: 99%

Generation of a Highly Biomimetic Organoid, Including Vasculature, Resembling the Native Immature Testis Tissue

Cham

Ibtisham

Fayaz

et al. 2021

Cells

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The creation of a testis organoid (artificial testis tissue) with sufficient resemblance to the complex form and function of the innate testis remains challenging, especially using non-rodent donor cells. Here, we report the generation of an organoid culture system with striking biomimicry of the native immature testis tissue, including vasculature. Using piglet testis cells as starting material, we optimized conditions for the formation of cell spheroids, followed by long-term culture in an air–liquid interface system. Both fresh and frozen-thawed cells were fully capable of self-reassembly into stable testis organoids consisting of tubular and interstitial compartments, with all major cell types and structural details expected in normal testis tissue. Surprisingly, our organoids also developed vascular structures; a phenomenon that has not been reported in any other culture system. In addition, germ cells do not decline over time, and Leydig cells release testosterone, hence providing a robust, tunable system for diverse basic and applied applications.

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Section: Discussionmentioning

confidence: 99%

Generation of a Highly Biomimetic Organoid, Including Vasculature, Resembling the Native Immature Testis Tissue

Cham

Ibtisham

Fayaz

et al. 2021

Cells

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“…As a substitution of serum, KSR was first used to maintain mouse embryonic stem cells (mESCs). Recently, researchers observed that KSR can promote the proliferation and differentiation of adipose-derived MSCs in monolayer cultures [31, 32] and facilitate the formation of 3D rat testicular culture, indicating that KSR seems to be a suitable substitution of FBS for 2D and 3D cell cultures [33]. To test the effect of KSR-containing medium on hMSC culture, we performed a systematical comparison of different hMSC culture media, including L-FBS, KSR containing MiPS, and optimized L-KSR (Figure 1(a)).…”

Section: Resultsmentioning

confidence: 99%

“…In fact, KSR is not only necessary but also sufficient to promote hMSC spheroid formation when added into different basal media (Figures 1(b)–1(d) and ). KSR is a substitute of FBS, and all the components are well defined, consisting of albumin, transferrin, insulin, collagen precursors, amino acids, vitamins, antioxidants, and trace elements [33]. Therefore, KSR is more suitable for clinical-grade MSC production as it eliminates many of the uncertainties encountered when using poorly defined serum supplements.…”

Section: Discussionmentioning

confidence: 99%

Serum-Free Culture System for Spontaneous Human Mesenchymal Stem Cell Spheroid Formation

Dong

Wang

et al. 2019

Stem Cells International

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Human mesenchymal stem cells (hMSCs) are widely used in clinical research because of their multipotential, immunomodulatory, and reparative properties. Previous studies determined that hMSC spheroids from a three-dimensional (3D) culture possess higher therapeutic efficacy than conventional hMSCs from a monolayer (2D) culture. To date, various 3D culture methods have been developed to form hMSC spheroids but most of them used culture medium containing fetal bovine serum (FBS), which is not suitable for further clinical use. Here, we demonstrate that dissociated single MSCs seeded in induced pluripotent stem medium (MiPS) adhere loosely to the dish and spontaneously migrate to form spheroids during day 3 to day 6. Through component deletion screening and complementation experiments, the knockout serum replacement (KSR) was identified as necessary and sufficient for hMSC spheroid formation. Transcriptome analysis showed that the overall expression profiles were highly similar between 2D culture with FBS and KSR-derived spheroids. Interestingly, genes related to inflammatory response, immune response, and angiogenesis were upregulated in spheroids at day 6 and qPCR results further validated the increased expression level of related genes, including STC1, CCL7, HGF, IL24, and TGFB3. When spheroids were replated in normal FBS medium, cells formed a typical spindle-shaped morphology and FACS results showed that the recovered cells retained MSC-specific surface markers, such as CD73, CD90, and CD105. In summary, we developed a practical and convenient method to generate hMSC spheroids for clinical research and therapy.

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“… 87 Sertoli cells, Leydig cells, and SSCs from rats were also co-cultured in the extracellular matrix to establish an in vitro toxicity test system for rat testes. 88 The three-dimensional culture system provides a research model for communication and interactions between cells in the body. However, it cannot sustain the culture of SSCs for a long period and is incapable of recycling all cells.…”

Section: Culture Of Sscsmentioning

confidence: 99%

Recent advances in isolation, identification, and culture of mammalian spermatogonial stem cells

Hao

Ren

et al. 2022

Asian J Androl

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Continuous spermatogenesis depends on the self-renewal and differentiation of spermatogonial stem cells (SSCs). SSCs, the only male reproductive stem cells that transmit genetic material to subsequent generations, possess an inherent self-renewal ability, which allows the maintenance of a steady stem cell pool. SSCs eventually differentiate to produce sperm. However, in an in vitro culture system, SSCs can be induced to differentiate into various types of germ cells. Rodent SSCs are well defined, and a culture system has been successfully established for them. In contrast, available information on the biomolecular markers and a culture system for livestock SSCs is limited. This review summarizes the existing knowledge and research progress regarding mammalian SSCs to determine the mammalian spermatogenic process, the biology and niche of SSCs, the isolation and culture systems of SSCs, and the biomolecular markers and identification of SSCs. This information can be used for the effective utilization of SSCs in reproductive technologies for large livestock animals, enhancement of human male fertility, reproductive medicine, and protection of endangered species.

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The use of KnockOut serum replacement (KSR) in three dimensional rat testicular cells co-culture model: An improved male reproductive toxicity testing system

Cited by 25 publications

References 25 publications

Generation of a Highly Biomimetic Organoid, Including Vasculature, Resembling the Native Immature Testis Tissue

Generation of a Highly Biomimetic Organoid, Including Vasculature, Resembling the Native Immature Testis Tissue

Serum-Free Culture System for Spontaneous Human Mesenchymal Stem Cell Spheroid Formation

Recent advances in isolation, identification, and culture of mammalian spermatogonial stem cells

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