Transcriptome profile of bovine iPSCs derived from Sertoli Cells

Jiang, Yu; An, Xing-Lan; Yu, Hao; Cai, Ningning; Zhai, Yanhui; Li, Qi; Cheng, Hui; Zhang, Sheng; Tang, Bo; Li, Ziyi; Zhang, Xueming

doi:10.1016/j.theriogenology.2019.11.022

Cited by 14 publications

(20 citation statements)

References 56 publications

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“…Animal procedures were approved by the Institutional Animal Care and Use Committee of Jilin University (SY201903002). The testes were processed as described (Cai, Tang, et al., 2016; Jiang, An, et al., 2020; Jiang, Zhu, et al., 2020; Wu et al., 2011). Briefly, the testicular parenchyma was cut into 0.3 to 0.5 cm 3 fragments and transferred into 5 ml cryovials containing freezing medium (10% DMSO + 2.5% newborn calf serum in MEM) after decapsulation.…”

Section: Methodsmentioning

confidence: 99%

“…To collect the seminiferous cords, the cryovials containing the cryopreserved tissues were swiftly transferred in a 37°C water bath and swirled for three minutes. The testicular tissues were processed as described (Cai, Tang, et al., 2016; Izadyar et al., 2002; Jiang, An, et al., 2020; Jiang, Zhu, et al., 2020; Wu et al., 2011). Briefly, the testicular fragments were washed with M 1 (MEM supplemented with 10% NCS + 2 mM glutamine + 100 IU/ml penicillin‐streptomycin + 40 µg/ml gentamycin + 15 mM HEPES), pipetted violently and allowed to sediment for three minutes.…”

Section: Methodsmentioning

confidence: 99%

“…To prepare feeder layer, bovine Sertoli cells were enriched and cultured in M 3 (DMEM supplemented with 10% FBS, 1% penicillin‐streptomycin and 1% NEAA) at 6 × 10 4 /cm 2 density in Φ 35 mm Petri dishes coated with 1% gelatin (Jiang, An, et al., 2020; Jiang, Zhu, et al., 2020). When Sertoli cells reach 50%–60% confluence, mitomycin C of 10 μg/ml was added into the medium.…”

Section: Methodsmentioning

confidence: 99%

“…Several markers have been reported in rodent and human undifferentiated spermatogonia (de Rooij, 2017; Guo et al., 2014; Kubota & Brinster, 2018); however, characterisation of bovine corresponding markers remains difficult due to lack of specific commercial antibodies. Recently, progresses have been made in this respect (Cai, Tang, et al., 2016; Goissis et al., 2018; Jiang, An, et al., 2020; Jiang, Zhu, et al., 2020; Kim et al., 2019). During the last decade, efforts have been made to optimise the culture system of bovine spermatogonia from different aspects including media and serum (Izadyar et al., 2003), feeder cells (Nasiri et al., 2012; Oatley et al., 2004), extracellular matrix (Akbarinejad et al., 2015; Kim et al., 2014), hormones (Narenji Sani et al., 2013) and growth factors (Aponte et al., 2006, 2008; Sahare et al., 2016).…”

Section: Introductionmentioning

confidence: 99%

“…Some small molecules were demonstrated to be effective for pluripotency maintenance and expansion of embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs) and mesenchymal stem cells (MSCs) (Hou et al., 2013; Jiang, An, et al, 2020; Zhao et al., 2018). The inhibitors of Mek1/2 (PD0325901) and Gsk3β (CHIR99021), known as 2i, have been used to culture ESCs and iPSCs (Jiang et al, 2019; Ware et al., 2014; Ying et al., 2008).…”

Section: Introductionmentioning

confidence: 99%

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Culture bovine prospermatogonia with 2i medium

et al. 2021

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Germplasm cryopreservation and expansion of gonocytes/prospermatogonia or spermatogonial stem cells (SSCs) are important; however, it's difficult in cattle. Since inhibitors of Mek1/2 and Gsk3β (2i) can enhance pluripotency maintenance, effects of 2i‐based medium on the cultivation of bovine prospermatogonia from the cryopreserved tissues were examined. The testicular tissues of newborn bulls were well cryopreserved. High mRNA levels of prospermatogonium/SSC markers (PLZF, GFRα‐1) and pluripotency markers (Oct4/Pouf5, Sox2, Nanog) were detected and the PLZF+/GFRα‐1+ prospermatogonia were consistently identified immunohistochemically in the seminiferous cords. Using differential plating and Percoll‐based centrifugation, 41.59% prospermatogonia were enriched and they proliferated robustly in 2i medium. The 2i medium boosted mRNA abundances of Pouf5, Sox2, Nanog, GFRα‐1, PLZF, anti‐apoptosis gene Bcl2, LIF receptor gene LIFR and enhanced PLZF protein expression, but suppressed mRNA expressions of spermatogonial differentiation marker c‐kit and pro‐apoptotic gene Bax, in the cultured prospermatogonia. It also alleviated H2O2‐induced apoptosis of the enriched cells and decreased histone H3 lysine (K9) trimethylation (H3K9me3) and its methylase Suv39h1/2 mRNA level in the cultured seminiferous cords. Overall, 2i medium improves the cultivation of bovine prospermatogonia isolated from the cryopreserved testes, by inhibiting Suv39h1/2‐mediated H3K9me3 through Mek1/2 and Gsk3β signalling, evidencing successful cryopreservation and expansion of bovine germplasm.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Culture bovine prospermatogonia with 2i medium

et al. 2021

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Comparative analysis of porcine iPSCs derived from Sertoli cells and fibroblasts

Zhu

Shen

et al. 2022

Journal Cellular Physiology

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Porcine embryonic fibroblasts (PEFs) can be directly reprogrammed into porcine induced pluripotent stem cells (piPSCs). However, the reprogramming process is generally lengthy and inefficient. Here, we established a fast and efficient induction system of piPSCs from porcine Sertoli cells (SCs) via forced expression of pig Yamanaka factors. The alkaline phosphatase (AP)-positive colonies from SCs developed on Day 3 after lentivirus infection, and were expanded and then picked up on Day 7, whereas reprogramming process from PEFs did not show any colonies in the same period. The picked piPSCs strongly expressed pluripotent genes, had the differentiation capacity to three germ layers, and could be also induced into primordial germ cell-like cells. Screening for transcription factor combinations showed that POU class 5 homeobox 1 (OCT4) is the core factor for AP-positive colony formation, and two factors (OCT4 and c-MYC) could successfully reprogram SCs into piPSCs. We then compared the RNA-sequencing data of piPSCs derived from SCs and PEFs, and found that the most significant difference was the activation of Transforming Growth Factor β signaling pathway. We also compared the RNA levels of SCs and PEFs, and found that SCs exhibited higher Wnt signaling activity and Bone Morphogenetic Protein 4 expression than PEFs, which might be correlated with higher cell proliferation rate and reprogramming efficiency. In summary, the data demonstrated that starting cell sources of piPSCs significantly affect reprogramming dynamics and SCs could serve as cell sources for efficient reprogramming.

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