Conversion of Primordial Germ Cells to Pluripotent Stem Cells: Methods for Cell Tracking and Culture Conditions

2013 ◽  
pp. 49-56 ◽  
Author(s):  
Go Nagamatsu ◽  
Toshio Suda
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Cell Tracking ◽  
Primordial Germ Cells ◽  
Culture Conditions

scholarly journals Formative pluripotent stem cells show features of epiblast cells poised for gastrulation

Cell Research ◽  
2021 ◽  
Author(s):  
Xiaoxiao Wang ◽  
Yunlong Xiang ◽  
Yang Yu ◽  
Ran Wang ◽  
Yu Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Primordial Germ Cells ◽  
Embryonic Stem ◽  
Large Set ◽  
Mouse Escs ◽  
Epiblast Stem Cells ◽  
Early Gastrula

AbstractThe pluripotency of mammalian early and late epiblast could be recapitulated by naïve embryonic stem cells (ESCs) and primed epiblast stem cells (EpiSCs), respectively. However, these two states of pluripotency may not be sufficient to reflect the full complexity and developmental potency of the epiblast during mammalian early development. Here we report the establishment of self-renewing formative pluripotent stem cells (fPSCs) which manifest features of epiblast cells poised for gastrulation. fPSCs can be established from different mouse ESCs, pre-/early-gastrula epiblasts and induced PSCs. Similar to pre-/early-gastrula epiblasts, fPSCs show the transcriptomic features of formative pluripotency, which are distinct from naïve ESCs and primed EpiSCs. fPSCs show the unique epigenetic states of E6.5 epiblast, including the super-bivalency of a large set of developmental genes. Just like epiblast cells immediately before gastrulation, fPSCs can efficiently differentiate into three germ layers and primordial germ cells (PGCs) in vitro. Thus, fPSCs highlight the feasibility of using PSCs to explore the development of mammalian epiblast.

BMP4 can generate primordial germ cells from bone-marrow-derived pluripotent stem cells

2012 ◽  
Vol 36 (12) ◽  
pp. 1185-1193 ◽  
Author(s):  
Reza Shirazi ◽  
Amir Hassan Zarnani ◽  
Masoud Soleimani ◽  
Mir Abbas Abdolvahabi ◽  
Karim Nayernia ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Primordial Germ Cells

scholarly journals 178 ESTABLISHMENT OF MOUSE PLURIPOTENT STEM CELLS GENERATED FROM PRIMORDIAL GERM CELLS

2005 ◽  
Vol 17 (2) ◽  
pp. 239
Author(s):  
S.W. Shim ◽  
S.J. Song ◽  
H.S. Shim ◽  
S.J. Uhm ◽  
B.Y. Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Lines ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Cultured Cells ◽  
Cell Layer ◽  
Es Cells ◽  
Primordial Germ Cells ◽  
Pcr Analysis ◽  
Feeder Cell

Pluripotent stem cells have been generated from two embryonic sources: ES cells generated from ICM of blastocyst stage embryos, and embryonic germ (EG) cells generated from primordial germ cells (PGCs). Both ES and EG cells are pluripotent and exhibit important characteristics such as high alkaline phosphatase (AP) activity, multicellular colony formation, normal and stable karyotype, continuous passaging ability, and capacity to differentiate into three embryonic germ layers. This study was performed to establish the culture system for mouse EG cells derived from mouse PGCs. PGCs collected from the genital ridge of Day 11.5, 12.5, and 13.5 mouse embryos (C57BL/6 × DBA/2) were cultured and subsequently passaged on mitotically inactivated STO feeder cell layer. Cells were grown in Dulbecco's modified eagle medium (DMEM) supplemented with 15% fetal bovine serum (FBS), 0.1 mM nonessential amino acids, 0.1 mM 2-mercaptoethanol, 2 mM glutamine, 100 IU/mL of penicillin, 100 μg/mL of streptomycin, 1,000 units/mL of leukemia inhibiting factor (LIF), 6 ng/mL of SCF, and 10 ng/mL of bFGF at culture conditions of 5% CO2 in air, 95% relative humidity, 37°C temperature. Cells were routinely passaged every 3–4 days. Over a period of 7–10 days in primary culture, PGCs proliferated to form small, densely packed, multicellular colonies consisting of AP-positive cells that morphologically resembled undifferentiated ES cells. RT-PCR analysis confirmed mRNA expression of transcription factors Oct-4 and Nanog in these cells. Cultured cells could be maintained on the feeder cell layer for at least 10 passages and still retain normal karyotype. These results suggest that cell lines derived from mouse primordial germ cells are presumably EG cell lines and could be useful for transgenic animal production and ES cell study.

scholarly journals Generation of Nanog reporter mice that distinguish pluripotent stem cells from unipotent primordial germ cells

genesis ◽  
2019 ◽  
Vol 57 (11-12) ◽  
Author(s):  
Maiko Terada ◽  
Masaki Kawamata ◽  
Ryota Kimura ◽  
Sayaka Sekiya ◽  
Go Nagamatsu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Primordial Germ Cells ◽  
Reporter Mice

An Evidence That Murine Marrow-Derived CXCR4+ SSEA-1+ Oct-4+ Very Small Embryonic-Like (VSEL) Stem Cells Are Pluripotent and Express Several Primordial Germ Cell (PGC) Markers - Hypotesis for Developmental Deposition of PGC in Various Organs.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1676-1676 ◽  
Author(s):  
Magda Kucia ◽  
Ewa Zuba-Surma ◽  
Ryan Reca ◽  
Janina Ratajczak ◽  
Mariusz Ratajczak
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Germ Cell ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Fetal Liver ◽  
Primordial Germ Cells ◽  
Embryonic Stem ◽  
Embryoid Bodies ◽  
C2c12 Cells

Abstract Recently we identified in murine BM a homogenous population of rare (~0.01% of BMMNC) Sca-1+ lin− CD45− cells that express by RQ-PCR and immunhistochemistry markers of pluripotent stem cells (PSC) such as SSEA-1, Oct-4, Nanog and Rex-1 and highly express Rif-1 telomerase protein (Leukemia2006;20,857–869). Direct electronmicroscopical analysis revealed that these cells display several features typical for embryonic stem cells such as i) small size (2–4 um in diameter), ii) large nuclei surrounded by a narrow rim of cytoplasm, and iii) open-type chromatin (euchromatin). We also found that VSELs may be released from BM and circulate in peripheral blood during tissue/organ injuries (e.g., heart infarct, stroke). Recently we noticed that ~5–10% of purified VSELs if plated over a C2C12 murine sarcoma cell feeder layer are able to form spheres that resemble embryoid bodies. Cells from these VSEL-derived spheres (VSEL-DS) are composed of immature cells with large nuclei containing euchromatin, and similarly as purified VSELs are CXCR4+SSEA-1+Oct-4+. Furthermore, VSEL-DS after replating over C2C12 cells may again (up to 5–7 passages) grow new spheres or if plated into cultures promoting tissue differentiation expand into cells from all three germ-cell layers. The formation of VSEL-DS was observed in a presence of C2C12 cells obtained from different sources. Furthermore, VSELs isolated from GFP+ mice grew GFP+ VSEL-DS which show a diploid content of DNA. This suggests that VSEL-DS are in fact derived from VSELs and not from the supportive C2C12 cell line as well as excludes the possibility of cell fusion to the observed phenomenon. Similar spheres were also formed by VSELs isolated from murine fetal liver, spleen and thymus. Interestingly formation of VSEL-DS was associated with a young age, and no VSEL-DS were observed by cells isolated from old mice (> 2 years). We also found that cells isolated from VSEL-DS similarly as embryonic stem cells grow tumors after injection into immunodeficient NOD/SCID mice (51/52 inoculated mice). Since VSELs isolated by us express several markers of primordial germ cells (fetal-type alkaline phosphatase, Oct-4, SSEA-1, CXCR4, Mvh, Stella, Fragilis, Nobox, Hdac6) we hypothesize that VSELs are closely related to a population of primordial germ cells. These cells are specified during early gastrulation in the proximal epiblast and subsequently migrate in a CXCR4-SDF-1 dependent manner through the embryo proper to their final destination in genital ridges. It is possible that some of these cells or a population of cells closely related to them migrate astray being chemoattracted by SDF-1 to fetal liver and subsequently, during the third trimester of gestation seed together with hematopoietic stem cells in bone marrow and perhaps other organs as well. In conclusion, we postulate that VSELs identified by us and purified at the single cell level could become an important source of pluripotent stem cells for regeneration.

scholarly journals Induction of Pluripotent Stem Cells from Primordial Germ Cells by Single Reprogramming Factors

Stem Cells ◽  
2013 ◽  
Vol 31 (3) ◽  
pp. 479-487 ◽  
Author(s):  
Go Nagamatsu ◽  
Takeo Kosaka ◽  
Shigeru Saito ◽  
Hiroaki Honda ◽  
Keiyo Takubo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Primordial Germ Cells ◽  
Reprogramming Factors

scholarly journals Generation of primordial germ cells from pluripotent stem cells

2009 ◽  
Vol 78 (2-3) ◽  
pp. 116-123 ◽  
Author(s):  
Cristina Eguizabal ◽  
Tanya C. Shovlin ◽  
Gabriela Durcova-Hills ◽  
Azim Surani ◽  
Anne McLaren
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Primordial Germ Cells

scholarly journals Distinct requirements for energy metabolism in mouse primordial germ cells and their reprogramming to embryonic germ cells

2017 ◽  
Vol 114 (31) ◽  
pp. 8289-8294 ◽  
Author(s):  
Yohei Hayashi ◽  
Kei Otsuka ◽  
Masayuki Ebina ◽  
Kaori Igarashi ◽  
Asuka Takehara ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Energy Metabolism ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Cell Function ◽  
Primordial Germ Cells ◽  
Embryonic Stem ◽  
Embryonic Germ Cells ◽  
Protein Levels

Primordial germ cells (PGCs), undifferentiated embryonic germ cells, are the only cells that have the ability to become gametes and to reacquire totipotency upon fertilization. It is generally understood that the development of PGCs proceeds through the expression of germ cell-specific transcription factors and characteristic epigenomic changes. However, little is known about the properties of PGCs at the metabolite and protein levels, which are directly responsible for the control of cell function. Here, we report the distinct energy metabolism of PGCs compared with that of embryonic stem cells. Specifically, we observed remarkably enhanced oxidative phosphorylation (OXPHOS) and decreased glycolysis in embryonic day 13.5 (E13.5) PGCs, a pattern that was gradually established during PGC differentiation. We also demonstrate that glycolysis and OXPHOS are important for the control of PGC reprogramming and specification of pluripotent stem cells (PSCs) into PGCs in culture. Our findings about the unique metabolic property of PGCs provide insights into our understanding of the importance of distinct facets of energy metabolism for switching PGC and PSC status.

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