Nanostructure and β1-integrin distribution analysis of pig's spermatogonial stem cell by atomic force microscopy

Li, Shengpu; Shi, Ruyi; Wang, Qiulan; Cai, Jiye; Zhang, Shouquan

doi:10.1016/j.gene.2011.12.044

Cited by 11 publications

(11 citation statements)

References 35 publications

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“…The structural information and mechanical properties of cell surface membranes are very important indicators for elucidating differentiation processes (Evans & Calderwood, ). We had tested the cellular membrane roughness of PSSCs in previous experiment (Li et al ., ), but without any other force index, such as stiffness force, adhesive force and Young's modulus which could reflect characteristics of PSSCs more completely. In this study, AFM were very useful in comparing the 3D morphology and other force features of undifferentiated and differentiated PSSCs.…”

Section: Discussionmentioning

confidence: 99%

“…Atomic force microscopy was performed as described previously to reveal the cellular ultrastructure and measures several cellular mechanical properties of the PSSCs (Li et al ., ). In brief, the cells were placed in an incubation solution (DMEM/F12 with 10% FBS, 1000 IU ml −1 LIF and 20 ng ml −1 GDNF, 10 ml) and then seeded onto clean coverslips for 3–5 min to allow for adherence; the coverslips were fixed with 2.5% glutaraldehyde for 15 min before use, washed with pure water for three times and air‐dried.…”

Section: Methodsmentioning

confidence: 97%

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Characteristics of spermatogonial stem cells derived from neonatal porcine testis

Shi

Bai

et al. 2014

Andrologia

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The aim of this study was to isolate and characterise porcine spermatogonial stem cells (PSSCs). The putative porcine germline stem cells from testis were isolated successfully by an improving way of enrichment with lymphocyte separation medium (LSM). Results from RT-PCR analyses showed that PSSCs were positive for OCT4, SOX2, NANOG, PGP9.5, c-MYC, KEL4 and PRDM-14 which are multipotent stem cell markers. At the protein level, the results of immunofluorescence analyses showed that PSSCs were positive for OCT4, PGP9.5, SOX2 and CD29. We successfully differentiated these PSSCs into adipocytes and muscle cells and then defined their characteristics, including morphology, surface stem cell markers, and mechanical properties. But the experiment of teratoma formation was negative. The results indicated the PSSCs could be multipotent. Atomic force microscopy was used to characterise the morphological and mechanical properties of undifferentiated PSSCs, as well as the differentiated adipocytes and muscle cells, which could be potentially useful for distinguishing PSSCs from differentiated cells.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 97%

Characteristics of spermatogonial stem cells derived from neonatal porcine testis

Shi

Bai

et al. 2014

Andrologia

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“…Oxidative stress related to the generation of reactive oxygen species (ROS) has also been associated with cell proliferation, differentiation and apoptosis in various systems (Dragin et al, 2006;Abrahan et al, 2009;Guo et al, 2010). Se is the catalytic center of several antioxidant enzymes and proteins such as glutathione peroxidase (GSH-Px) and selenoprotein P, some of which are involved in the regulation of oxidative microenvironment (Rotruck et al, 1973;Carlson et al, 2009;Wayne and Zeynep, 2010;Li et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Effects of different levels of dietary selenium on the proliferation of spermatogonial stem cells and antioxidant status in testis of roosters

Shi

Zhao

Ren

et al. 2014

Animal Reproduction Science

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“…These shortcomings could be addressed by identification of SSC markers and also by examining the factors that regulate the fate of SSCs during in vitro culture. Although putative SSC markers, such as GDNF family receptor α-1 (GFRα-1), α 6 β 1 -integrins, epithelial cell adhesion molecule (EpCAM), promyelocytic leukemia zinc finger (PLZF; ZBTB16), thymus cell antigen-1 (Thy-1, CD90), LIN28, E-cadherin type 1 (CDH1), POU domain class 5 homeobox 1 (POU5F1) and Nanos 2 and 3 are promising candidates for purification and characterization of SSCs in a number of species [9,10,11,12,13,14,15,16,17,18,19], the definite characterization of “true” SSCs is confirmed if these cells can colonize and produce sperm following transplantation into the recipient's seminiferous tubules [5]. To date, GFRα-1 receptor is mostly used as a consensus marker for SSC identification in several species, including rodents [20].…”

mentioning

confidence: 99%

Characterization and <i>In Vitro</i> Culture of Putative Spermatogonial Stem Cells Derived from Feline Testicular Tissue

et al. 2013

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Spermatogonial stem cells (SSCs) function to regulate the balance of self-renewal and differentiation of male gametes. SSCs have been successfully isolated and cultured in vitro in several species, but not in feline. Therefore, in this study, we aimed to culture and characterize feline SSCs. In experiment 1, testes (n=5) from different pubertal domestic cats were cryosectioned and fluorescently immunolabeled to examine the expression of SSC (GFRα-1), differentiated spermatogonium (c-kit) and germ cell (DDX-4) markers. In experiments 2 and 3, testicular cells were digested and subsequently cultured in vitro. The resultant presumptive SSC colonies were then collected for SSC identification (experiment 2), or further cultured in vitro on feeder cells (experiment 3). Morphology, gene expression and immunofluorescence were used to identify the SSCs. Experiment 1 demonstrated that varying types of spermatogenic cells existed and expressed different germ cell/SSC markers. A rare population of putative SSCs located at the basement membrane of the seminiferous tubules was specifically identified by co-expression of GFRα-1 and DDX-4. Following enzymatic digestion, grape-like colonies formed by 13-15 days of culture. These colonies expressed GFRA1 and ZBTB16, but did not express KIT. Although we successfully isolated and cultured feline SSCs in vitro, the SSCs could only be maintained for 57 days. In conclusion, this study demonstrates, for the first time, that putative SSCs from testes of pubertal domestic cats can be isolated and cultured in vitro. These cells exhibited SSC morphology and expressed SSC-specific genes. However, long-term culture of these putative SSCs was compromised.

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Nanostructure and β1-integrin distribution analysis of pig's spermatogonial stem cell by atomic force microscopy

Cited by 11 publications

References 35 publications

Characteristics of spermatogonial stem cells derived from neonatal porcine testis

Characteristics of spermatogonial stem cells derived from neonatal porcine testis

Effects of different levels of dietary selenium on the proliferation of spermatogonial stem cells and antioxidant status in testis of roosters

Characterization and <i>In Vitro</i> Culture of Putative Spermatogonial Stem Cells Derived from Feline Testicular Tissue

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