Isolation of novel developmental genes from human germ cell, oocyte and embryo cDNA by differential display

2001 ◽  
Vol 13 (1) ◽  
pp. 51 ◽  
Author(s):  
Marilyn Monk ◽  
Cathy Holding ◽  
Tetsuya Goto
Keyword(s):  
Stem Cell ◽  
Early Development ◽  
Germ Cells ◽  
Differential Display ◽  
Primordial Germ Cells ◽  
Stem Cell Differentiation ◽  
Human Reproduction ◽  
Human Embryos ◽  
Embryonic Cells ◽  
Developmental Genes

Due to the difficulties inherent in research on human embryos, almost nothing is known about genes active in human early development. Although the human genome project will provide resources that theoretically provide access to every human gene, those genes specific to human early development may be difficult to define. Also, by definition, genes specific to early development will not be represented in cDNA databases derived from human somatic cells. Yet these unknown human developmental genes are likely to be of key importance for several areas of human health, including assisted reproduction and contraception, embryo stem cell research and tissue trans-plantation, ageing and cancer. In order to identify and isolate these human developmental genes, we have prepared amplified cDNA from human primordial germ cells, oocytes and embryos, and used differential display to compare patterns of gene expression in these embryonic cells and in the cells of somatic tissues of a 10-week human fetus. This paper reviews the highly sensitive procedures used to create amplified cDNA representing expressed genes in a single cell and the use of differential display to identify developmental genes. Several such genes have been isolated, but their full-length sequences and function are yet to be elucidated. Genes active in human early development are expected to play key roles in the maintenance of the archetypal stem cell state, potential immortality and the invasiveness of trophectoderm and primordial germ cells. They represent candidate genes regulating these functions for targeting in clinical research in human reproduction, stem cell differentiation and cancer.

scholarly journals Development and Conservation of Gonadal Primordial Germ Cells for Preservation of Local Chicken in Indonesia

2017 ◽  
Vol 26 (3) ◽  
pp. 125 ◽  
Author(s):  
Tatan Kostaman ◽  
S Sopiyana
Keyword(s):  
Germ Cell ◽  
Early Development ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Primordial Germ Cell ◽  
Germplasm Conservation ◽  
Ex Situ ◽  
Management Systems ◽  
Embryonic Cells ◽  
Unique Nature

One of the <em>ex situ</em> conservation techniques for poultry that recently developed was to collect primordial germ cell (PGC) or gonadal primordial germ cell (gPGC) that isolated from embryo development. Primordial germ cells (PGC) are embryonic cells that migrate to the gonads and form the precursors of gametes. The unique nature and accessibility of PGC during the early development provides an opportunity to manipulate the poultry germplasm, for example by forming germline chimeras. There are some stages that must be done through isolation and collection of PGC from its resources i.e. blastoderm, embryonic circulation blood and gonad. PGC collection originating from the gonads is one of existing PGC resources and technologies. gonadal PGC have advantages compared with other sources, namely (1) A large number of gonadal PGC can be taken from an embryo; and (2) A collection of gonadal PGC can be used in developing management systems of local avian germplasm conservation. This review is intended to describe the usefulness of isolation and collection technology of gonadal PGC as the local poultry germplasm conservation in Indonesia.

Stem cell factor protects germ cells from apoptosis in vitro

2000 ◽  
Vol 113 (1) ◽  
pp. 161-168 ◽  
Author(s):  
W. Yan ◽  
J. Suominen ◽  
J. Toppari
Keyword(s):  
Stem Cell ◽  
Germ Cells ◽  
Stem Cell Factor ◽  
Primordial Germ Cells ◽  
Survival Function ◽  
Seminiferous Tubules ◽  
Testicular Development ◽  
Dna Laddering ◽  
Survival Effect

Stem cell factor (SCF) plays an important role in migration, adhesion, proliferation, and survival of primordial germ cells and spermatogonia during testicular development. However, the function of SCF in the adult testis is poorly described. We have previously shown that, in the presence of SCF, there were more type A spermatogonia incorporating thymidine at stage XII of rat seminiferous tubules cultured in vitro than in the absence of SCF, implying that the increased DNA synthesis might result from enhanced survival of spermatogonia. To explore the potential pro-survival function of SCF during spermatogenesis, the seminiferous tubules from stage XII were cultured in the presence or absence of SCF (100 ng/ml) for 8, 24, 48, and 72 hours, respectively, and apoptosis was analyzed by DNA laddering and in situ 3′-end labeling (ISEL) staining. Surprisingly, not only spermatogonia, but also spermatocytes and spermatids, were protected from apoptosis in the presence of SCF. Apoptosis took place much later and was less severe in the SCF-treated tubules than in the controls. Based on previous studies showing that FSH prevents germ cells from undergoing apoptosis in vitro, and that SCF level is increased dramatically in response to FSH stimulation, we also tested if the pro-survival effect of FSH is mediated through SCF by using a function-blocking monoclonal antibody, ACK-2, to block SCF/c-kit interaction. After 24 hours of blockade, the protective effect of FSH was partially abolished, as manifested by DNA laddering and ISEL analyses. The present study demonstrates that SCF acts as an important survival factor for germ cells in the adult rat testis and FSH pro-survival effect on germ cells is mediated partially through the SCF/c-kit pathway.

Early development of primordial germ cells in Pacific bluefin tuna Thunnus orientalis

2019 ◽  
Vol 131 ◽  
pp. 106-112 ◽  
Author(s):  
Kentaro Higuchi ◽  
Rie Goto ◽  
Junpei Konishi ◽  
Yoshiaki Ina ◽  
Yukinori Kazeto ◽  
...  
Keyword(s):  
Early Development ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Bluefin Tuna ◽  
Pacific Bluefin Tuna ◽  
Thunnus Orientalis

Role of stem cell factor in somatic–germ cell interactions during prenatal oogenesis

Zygote ◽  
1996 ◽  
Vol 4 (04) ◽  
pp. 349-351 ◽  
Author(s):  
Massimo De Felici ◽  
Anna Di Carlo ◽  
Maurizio Pesce
Keyword(s):  
Stem Cell ◽  
Germ Cells ◽  
Stem Cell Factor ◽  
Molecular Mechanisms ◽  
Primordial Germ Cells ◽  
Significant Progress ◽  
Proliferation And Differentiation ◽  
Complex Events ◽  
Mechanisms Of Migration

During embryogenesis germ cells originate from primordial germ cells (PGCs). The development of mammalian PGCs involves a number of complex events (formation and segregation of PGC precursors, PGC migration and proliferation) which lead to the differentiation of oocytes or prospermatogonia (for a review see De Feliciet al., 1992). During recent years developments in methods for isolation, purification and culture of mouse PGCs have led to significant progress in the understanding of molecular mechanisms of migration, proliferation and differentiation of these cells (for reviews see De Felici, 1994; and De Felici &amp; Pesce, 1994a). In this paper we describe the key role played by stem cell factor (SCF) in PGC development and early folliculogenesis.

Primary culture of porcine PGCs requires LIF and porcine membrane-bound stem cell factor

Zygote ◽  
1998 ◽  
Vol 6 (3) ◽  
pp. 271-275 ◽  
Author(s):  
Gabriela Durcova-Hills ◽  
Katja Prelle ◽  
Sigrid Müller ◽  
Miodrag Stojkovic ◽  
Jan Motlik ◽  
...  
Keyword(s):  
Stem Cell ◽  
Primary Culture ◽  
Germ Cells ◽  
Stem Cell Factor ◽  
Primordial Germ Cells ◽  
Embryonic Stem ◽  
Leukaemia Inhibitory Factor ◽  
Feeder Cells ◽  
Membrane Bound

We studied the effect of murine leukaemia inhibitory factor (LIF), human basic fibroblast growth factor (bFGF) and porcine stem cell factor (SCF) on the survival and/or proliferation of porcine primordial germ cells (PGCs) obtained from 27-day-old embryos in vitro. PGCs were cultured in embryonic stem cell (ESC) medium supplemented with or without either LIF (1000 IU/ml) alone or LIF together with bFGF (10 ng/ml). They were seeded on mitotically inactivated feeder cells, either STO or transfected STO cells (STO#8), expressing the membrane-bound form of porcine SCF. PGCs were identified by their alkaline phosphatase (AP) activity and counted after 1, 3 and 5 days in culture. After 1 day of culture, PGCs cultured on STO#8 cells showed significantly higher survival than PGCs cultured on STO cells (p < 0.05). The combined effect of SCF and LIF caused a significant increase in PGC number by day 3 of culture when PGCs were cultured on either STO cells (p < 0.01) or STO#8 (p < 0.001). When SCF and LIF were used together with bFGF no increase in the PGC number was observed. Our results suggest that the membrane-bound form of porcine SCF plays a pivotal role in the primary culture of porcine PGCs and that bFGF is not required in vitro.

scholarly journals HIPSTR and thousands of lncRNAs are heterogeneously expressed in human embryos, primordial germ cells and stable cell lines

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Dinar Yunusov ◽  
Leticia Anderson ◽  
Lucas Ferreira DaSilva ◽  
Joanna Wysocka ◽  
Toshihiko Ezashi ◽  
...  
Keyword(s):  
Cell Lines ◽  
Germ Cells ◽  
Expression Patterns ◽  
Primordial Germ Cells ◽  
Population Level ◽  
Human Embryos ◽  
Cell Stage ◽  
Lncrna Gene ◽  
Stable Cell Lines ◽  
The Individual

Abstract Eukaryotic genomes are transcribed into numerous regulatory long non-coding RNAs (lncRNAs). Compared to mRNAs, lncRNAs display higher developmental stage-, tissue-, and cell-subtype-specificity of expression, and are generally less abundant in a population of cells. Despite the progress in single-cell-focused research, the origins of low population-level expression of lncRNAs in homogeneous populations of cells are poorly understood. Here, we identify HIPSTR (Heterogeneously expressed from the Intronic Plus Strand of the TFAP2A-locus RNA), a novel lncRNA gene in the developmentally regulated TFAP2A locus. HIPSTR has evolutionarily conserved expression patterns, its promoter is most active in undifferentiated cells, and depletion of HIPSTR in HEK293 and in pluripotent H1BP cells predominantly affects the genes involved in early organismal development and cell differentiation. Most importantly, we find that HIPSTR is specifically induced and heterogeneously expressed in the 8-cell-stage human embryos during the major wave of embryonic genome activation. We systematically explore the phenomenon of cell-to-cell variation of gene expression and link it to low population-level expression of lncRNAs, showing that, similar to HIPSTR, the expression of thousands of lncRNAs is more highly heterogeneous than the expression of mRNAs in the individual, otherwise indistinguishable cells of totipotent human embryos, primordial germ cells, and stable cell lines.

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