Fractionation of human spermatogenic cells using STA-PUT gravity sedimentation and their miRNA profiling

Liu, Yun; Niu, Minghui; Yao, Chencheng; Hai, Yanan; Yuan, Quan; Liu, Yang; Guo, Ying; Li, Zheng; He, Zuping

doi:10.1038/srep08084

Cited by 85 publications

(75 citation statements)

References 45 publications

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“…Gene expression and chromatin state in male germ cells 59 We collected H3K4me3 and H3K27me3 ChIP-seq data, as well as RNA-seq data, in sorted male 60 germ cells from five species spanning 175 million years of evolutionary divergence in the 61 mammalian lineage: human, rhesus macaque, mouse, bull, and opossum ( Table 1). We obtained data from cells at two time points during male 63 germ cell development: prophase of meiosis I (pachytene spermatocytes), and following 64 completion of meiosis (round spermatids).…”

mentioning

confidence: 99%

Parallel evolution of male germline epigenetic poising and somatic development in animals

et al. 2016

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confidence: 99%

Parallel evolution of male germline epigenetic poising and somatic development in animals

et al. 2016

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“…Nevertheless, it remains to be defined whether miRNAs are required for spermatogenesis in humans. Recently, we have identified 559 miRNAs distinctively expressed by human spermatogonia, pachytene spermatocytes and round spermatids (Liu et al 2015). Using miRNA microarray, we have identified 144 miRNAs that are significantly up-regulated while 29 miRNAs are downregulated in pachytene spermatocytes vs round spermatids (Liu et al 2015), reflecting essential roles of these miRNAs in mediating spermiogenesis.…”

Section: Functions Of Mirnas In Meiosis and Spermiogenesismentioning

confidence: 99%

“…Recently, we have identified 559 miRNAs distinctively expressed by human spermatogonia, pachytene spermatocytes and round spermatids (Liu et al 2015). Using miRNA microarray, we have identified 144 miRNAs that are significantly up-regulated while 29 miRNAs are downregulated in pachytene spermatocytes vs round spermatids (Liu et al 2015), reflecting essential roles of these miRNAs in mediating spermiogenesis. A number of novel binding targets of the differentially expressed miRNAs have been identified using various bioinformatics software and verified by real time PCR (Liu et al 2015).…”

Section: Functions Of Mirnas In Meiosis and Spermiogenesismentioning

confidence: 99%

“…SSCs, spermatogonial stem cells; Spc, spermatocytes; RS, round spermatids; ES, elongating spermatids. (Liu et al 2015), suggesting that these miRNAs are involved in mitosis and meiosis.…”

Section: The Roles Of Mirnas In Ssc Self-renewal and Differentiationmentioning

confidence: 99%

“…Using miRNA microarray, we have identified 144 miRNAs that are significantly up-regulated while 29 miRNAs are downregulated in pachytene spermatocytes vs round spermatids (Liu et al 2015), reflecting essential roles of these miRNAs in mediating spermiogenesis. A number of novel binding targets of the differentially expressed miRNAs have been identified using various bioinformatics software and verified by real time PCR (Liu et al 2015). Significantly, our ability to unveil the global distinct miRNA signatures and binding targets of human spermatogonia, pachytene spermatocytes and round spermatids could provide novel small RNA regulatory mechanisms mediating three stages of human spermatogenesis and offers new targets for treatment of male infertility.…”

Section: Functions Of Mirnas In Meiosis and Spermiogenesismentioning

confidence: 99%

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MicroRNAs and DNA methylation as epigenetic regulators of mitosis, meiosis and spermiogenesis

Yao¹,

Liu²,

Sun³

et al. 2015

Reproduction

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Spermatogenesis is composed of three distinctive phases, which include self-renewal of spermatogonia via mitosis, spermatocytes undergoing meiosis I/II and post-meiotic development of haploid spermatids via spermiogenesis. Spermatogenesis also involves condensation of chromatin in the spermatid head before transformation of spermatids to spermatozoa. Epigenetic regulation refers to changes of heritably cellular and physiological traits not caused by modifications in the DNA sequences of the chromatin such as mutations. Major advances have been made in the epigenetic regulation of spermatogenesis. In this review, we address the roles and mechanisms of epigenetic regulators, with a focus on the role of microRNAs and DNA methylation during mitosis, meiosis and spermiogenesis. We also highlight issues that deserve attention for further investigation on the epigenetic regulation of spermatogenesis. More importantly, a thorough understanding of the epigenetic regulation in spermatogenesis will provide insightful information into the etiology of some unexplained infertility, offering new approaches for the treatment of male infertility.Reproduction (2015) 150 R25-R34

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Simultaneous Purification of Round and Elongated Spermatids from Testis Tissue Using a FACS‐Based DNA Ploidy Assay

et al. 2018

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Spermiogenesis is the final phase of spermatogenesis during which post‐meiotic haploid round spermatids (rSpt) differentiate into elongated spermatozoa and includes several critical cell‐specific processes like DNA condensation, formation of the acrosome, and production of the flagellum. Disturbances in this process will lead to complications in sperm development and subsequently cause infertility. As such, studying spermiogenesis has clinical relevance in investigating the etiology of male infertility and will improve our scientific understanding of male germ cell formation. Here, we were able to purify round spermatid and elongated spermatid fractions from a single cryopreserved human testicular tissues sample with an efficiency of 85.4% ± 4.9% and 97.6% ± 0.6%, respectively. We confirmed the cell types by morphology and immunohistochemistry for histone H4 and PNA protein expression. The purity was measured by manual counting of histone H4 positive (round) and negative (elongated) spermatids in both sorted 1 N cell fractions. This method can be applied to both human and rodent studies. Especially in studies with limited access to testicular tissue, this method provides a reliable means to simultaneously isolate these cell types with high purity. Our method allows for further investigation of germ cell development and the process of spermiogenesis in particular, as well as provides a tool to study the etiology of male infertility, including morphological and biochemical assessment of round and elongating spermatids from subfertile men. © 2018 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

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Fractionation of human spermatogenic cells using STA-PUT gravity sedimentation and their miRNA profiling

Cited by 85 publications

References 45 publications

Parallel evolution of male germline epigenetic poising and somatic development in animals

Parallel evolution of male germline epigenetic poising and somatic development in animals

MicroRNAs and DNA methylation as epigenetic regulators of mitosis, meiosis and spermiogenesis

Simultaneous Purification of Round and Elongated Spermatids from Testis Tissue Using a FACS‐Based DNA Ploidy Assay

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