Identification of pluripotent cells in bovine uterus: in situ and in vitro studies

Łupicka, M.; Bodek, Gabriel; Shpigel, Nahum Y.; Elnekave, Ehud; Korzekwa, Anna

doi:10.1530/rep-14-0348

Cited by 24 publications

(26 citation statements)

References 35 publications

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“…Differentiation assays of adipogenic and osteogenic lineages of bovine eMSCs were confirmed in all samples with excellent response after staining (Figure ). We reinforce the results of Łupicka et al (,) which showed that uterine stromal cells differentiated to adipogenic, chondrogenic, and osteogenic lineage upon specific condition, confirming the wide plasticity of these source. We reached great response to differentiation once Donofrio et al () demonstrated the osteogenic potential of endometrial stromal cells and Cabezas et al () the osteogenic and chondrogenic potential of endometrial putative mesenchymal progenitor cells from late luteal phase.…”

Section: Resultssupporting

confidence: 92%

Bovine endometrial cells: a source of mesenchymal stem/progenitor cells

Moraes

Maia

Dias

et al. 2016

Cell Biology International

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Endometrial mesenchymal stem/progenitor cells (eMSCs) are multipotent cells known to modulate the immune system and have clinical application for human and animal health. This makes these bovines cells attractive for dual use as cellular therapy and experimental model. The aim of this study was to isolate, evaluate the differentiation potential, immunophenotypic and immunocytochemistry characteristics, chromosomal stability, cloning efficiency, and cryopreservation response of bovine eMSCs collected in two phases of the estrous cycle. For this, cells were isolated and submitted to differentiation for adipogenic and osteogenic lineage. The cells were then characterized by flow cytometer (FC) (vimentin, CD29, CD44, MHC-II, CD34) and immunocytochemistry (vimentin, pan-cytokeratin, CD44) and submitted to cytogenetic and cloning efficiency assay. The cells were also cryopreserved using two different medium of cryopreservation and analyzed by FC for viability, necrosis, late-apoptosis + necrosis, and initial apoptosis rates before and after cryopreservation. We obtained homogeneous cell populations which have fibroblastic morphology and adherence to plastic. These cells expressed high levels of markers CD29, CD44, and vimentin, low expression levels for CD34 and no MHC-II. The cells were chromosomally stable (2n = 60) with high cloning efficiency and no difference (P > 0.05) between medium of cryopreservation or phase was observed after thawing. We showed the presence and differentiation potential of bovine eMSCs, with chromosomal stability and great response to cryopreservation with both medium, which has implications for build biobanks or development of new therapeutic approaches to combat uterine diseases or to study.

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Section: Resultssupporting

confidence: 92%

Bovine endometrial cells: a source of mesenchymal stem/progenitor cells

Moraes

Maia

Dias

et al. 2016

Cell Biology International

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“…There are reports of the joint expression of OCT4 with SOX2 and NANOG in human endometrium, myometrium and endometriotic tissues (Götte et al., ), and in cultured stromal cells of porcine endometrium (Bodek et al., ). In bovine, only one study has shown Nanog expression associated with Oct4 and Sox2, in epithelial, stromal and myometrial cells between days 8 and 10 of the oestrous cycle, being highest mRNAs in the ipsilateral horn, suggesting the influence of ovarian hormones, and also revealing a decrease in pluripotent transcription factor expression in older cows (Łupicka et al., ).…”

Section: Discussionmentioning

confidence: 99%

Characterization of mesenchymal stem cells in bovine endometrium during follicular phase of oestrous cycle

Lara

Rivera

Rojas

et al. 2017

Reprod Domestic Animals

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Stem cells have been postulated as responsible for cell regeneration in highly and continuously regenerative tissues such as the endometrium. Few studies in cattle have identified and specified the presence of stem cells in the endometrium during the oestrous cycle. The aim of this study was to investigate the presence of mesenchymal stem cells (MSCs) in the bovine endometrium during the follicular phase (FP) of the oestrous cycle. Uterine tissue was collected in the time-frame comprising day 18 of the cycle and ovulation (day 0). We isolated, cultured and expanded four primary cell lines from endometrium and identified byRT-qPCR the expression of OCT4, SOX2 but not NANOG (undifferentiated/embryonic markers), CD44 (MSCs marker) and c-KIT (stem cell marker) genes; and the encoded Oct4, Sox2 and Cd44 proteins by Western blot or immunostaining of paraffin-embedded tissue in endometrium. We demonstrated that cells isolated from bovine endometrium displayed essentially the same gene expression pattern; however, at the protein level, Oct4 and Cd44 were not detected. Besides, they showed typical functional characteristics of MSCs such as fibroblast-like morphology, plastic adherence, high proliferative capacity, clone formation in vitro and the ability to differentiate into chondrogenic, osteogenic and adipogenic lineages. We obtained for the first time an extensive characterization of undifferentiated cells populations contained in the bovine endometrium during the FP of the oestrous cycle.

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“…Stem cells have been reported in human myometrial side population by others also [15] as a relatively quiescent population with majority of them being in G0 phase with low expression of ER & PR, grow under hypoxic conditions and abundant expression of OCT-4 & NANOG. Lupicka et al [16, 17] have reported presence of pluripotent cells expressing OCT-4, NANOG and SOX2 in all the compartment of bovine uterus including myometrium and also in bovine uterus with adenomyosis. Label retaining cells have been reported in beta-catenin deficient mouse uterine myometrium (degenerative uterus with myometrium replaced by adipose tissue) and they also respond to gonadotropins [18].…”

Section: Discussionmentioning

confidence: 99%

Very small embryonic-like stem cells (VSELs) in adult mouse uterine perimetrium and myometrium

Bhartiya

James

2017

J Ovarian Res

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We have earlier reported the presence of very small embryonic-like stem cells (VSELs) in adult mouse uterus along with slightly bigger progenitors termed endometrial stem cells (EnSCs) and their regulation by ovarian hormones thus demonstrating a crucial role played by them during proliferation, differentiation and remodeling of the endometrium. Present study is a brief communication wherein we have examined the effect of higher dose of estrogen (E, 2 μg/day), progesterone (P, 1 mg/day) and follicle stimulating hormone (FSH, 5 IU/day for 5 days) specifically on the myometrium and perimetrium surrounding the endometrium in bilaterally ovariectomized mice. Similar treatment with E & P was recently used in a study published in the journal Nature to study the effect of steroid hormones on hematopoietic stem cells and this treatment regimen helps achieve hormone levels observed during pregnancy. Quiescent spherical stem cells (lacking PCNA expression) with high nucleo-cytoplasmic ratio and nuclear OCT-4A were detected in the perimetrium of atrophied (bilaterally ovariectomized) uterus. PCNA expression was observed after treatment and cells with cytoplasmic OCT-4B were invariably observed in the myometrium. VSELs were clearly visualized after treatment and the effect of P and FSH was more prominent compared to E on the development of myometrium. It is speculated that stem cells with nuclear OCT-4A located in the perimetrium differentiate to give rise to endothelial and myometrial cells with cytoplasmic OCT-4B. Based on the results of present study and published reports showing the presence of pluripotent markers (OCT-4, NANOG and SOX2) in human myometrial side population and expression of particularly OCT-4A in human leiomyomas, we speculate that these nuclear OCT-4 positive stem cells located in the perimetrium are the possible tumor initiating cells leading to the development of leiomyomas rather than the mesenchymal cells which express cytoplasmic OCT-4B.

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Identification of pluripotent cells in bovine uterus: in situ and in vitro studies

Cited by 24 publications

References 35 publications

Bovine endometrial cells: a source of mesenchymal stem/progenitor cells

Bovine endometrial cells: a source of mesenchymal stem/progenitor cells

Characterization of mesenchymal stem cells in bovine endometrium during follicular phase of oestrous cycle

Very small embryonic-like stem cells (VSELs) in adult mouse uterine perimetrium and myometrium

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