147 Dynamics of drake spermatogenesis

2020 ◽  
Vol 32 (2) ◽  
pp. 200
Author(s):  
L. A. Volkova ◽  
A. N. Vetokh ◽  
E. K. Tomgorova ◽  
N. A. Volkova ◽  
H. V. Ashraf ◽  
...  
Keyword(s):  
Stem Cells ◽  
Genetic Material ◽  
Cell Types ◽  
Seminiferous Tubules ◽  
Age Group ◽  
Spermatogenic Cells ◽  
Russian Science ◽  
Donor Cells ◽  
Development And Differentiation ◽  
Species Specific

The examination of age dynamics for the development and differentiation of spermatogenic cells is of great importance to the study of spermatogenesis in poultry. Testicular stem cells, represented by spermatogonia, are a valuable genetic material for creating cryobanks of biomaterial. This is especially important when preserving and maintaining the gene pool of valuable breeds of poultry. In the process of differentiation, these cells give rise to a significant population of germ cells, so they can be used as donor cells for transplantation into the testes of male recipients. Thus, understanding species-specific characteristics of spermatogenesis in males is an important step for obtaining the spermatogenic cell population of interest. The aim of this research was to study the dynamics of spermatogenesis in drake. For the study, 10 groups of males were formed depending on age: 2 weeks and 1, 1.5, 2, 2.5, 3, 4, 5, 6, and 7 months. There were eight males in each age group from 2 weeks to 6 months and six males in the 7-month age group. The testes were isolated postmortem, fixed in Bouin's fixative solution, and embedded in paraffin, and histological sections (5µm) were cut. The following indicators were evaluated: diameter of the seminiferous tubules, types of spermatogenic cells in the seminiferous tubules, and the amount of these cells within seminiferous tubules. Statistical analysis was performed using a t-test in SPSS ver. 15.0 (IBM Corp.). The types of spermatogenic cells were identified by morphology, and no fewer than 30 seminiferous tubules were examined from each individual. The diameter of the seminiferous tubules in the drake testes increased with age. At the ages of 2 weeks and 1, 1.5, 2, 2.5, 3, 4, 5, 6, and 7 months, these indicators were 38±2, 55±2, 60±2, 61±3, 62±2, 62±4, 65±2, 76±3, 94±5, and 163±7µm, respectively. This was due to an increase in the number of spermatogenic cells within the seminiferous tubules to 23±1, 27±1, 38±1, 44±2, 47±3, 57±2, 68±2, 140±5, 187±7, and 466±13 at the ages of 2 weeks and 1, 1.5, 2, 2.5, 3, 4, 5, 6, and 7 months, respectively. The presence, number, and ratio of the cell populations varied depending on age. At the ages of 1-12 weeks, the main cell types in the seminiferous tubules were Sertoli cells and spermatogonia. After the age of 4 months, primary spermatocytes began to appear in the seminiferous tubules. Secondary spermatocytes were visualised at 5 months of age, whereas spermatids could be detected at 6 months of age. Mature sperm cells were detected in the seminiferous tubules of drakes at the age of 7 months. Based on the data obtained, the following conclusion can be made: from 1-12 weeks of age, the generative cells of the seminiferous tubules in drakes are represented mainly by spermatogonia (P<0.05). Therefore, this period can be considered optimal for obtaining testicular stem cells and carrying out manipulations with them. This study was supported by the Russian Science Foundation within project no.16-16-04104.

143 The Dynamics of Spermatogenesis in Guinea Fowls

2018 ◽  
Vol 30 (1) ◽  
pp. 211
Author(s):  
N. A. Volkova ◽  
A. N. Vetokh ◽  
I. P. Novgorodova ◽  
A. V. Dotsev ◽  
N. A. Zinovieva
Keyword(s):  
Germ Cells ◽  
Sertoli Cells ◽  
Genetic Material ◽  
Guinea Fowl ◽  
Seminiferous Tubules ◽  
Spermatogenic Cells ◽  
Russian Science ◽  
Effective Selection ◽  
Generative Cells ◽  
Selection Of

Male gonads are valuable genetic material for creation of biomaterial cryobanks to preserve the genes of various animals, including poultry. Spermatogonia, which are stem cells of the testes, are of greatest interest. For effective selection of spermatogenic cells, including spermatogonia, it is necessary to know the specific features of spermatogenesis of the species of interest. In this regard, the aim of this study was to investigate the dynamics of spermatogenesis in guinea fowl. Histological examinations of guinea fowl testes (n = 90 birds) were done for 9 age categories, from 2 wk to 6 months. For each individual, at least 30 seminiferous tubules were examined. Seminiferous tubule diameters and numbers and types of spermatogenic cells (based on morphology) were determined. Overall, the histologic structure of guinea fowl testes was similar to that of mammals. Cell populations of the seminiferous tubules included Sertoli cells and generative cells, including spermatogonia, spermatocytes, spermatids, and sperm, at various stages of differentiation. Diameter of seminiferous tubules was (mean ± SEM) 36 ± 1, 58 ± 1, 64 ± 1, 65 ± 1, 110 ± 3, 178 ± 4, 233 ± 4, 274 ± 6, and 295 ± 5 µm at 2 wk, 1, 1.5, 2, 2.5, 3, 4, 5, and 6 months, respectively. Furthermore, at those ages, the number of spermatogenic cells per tubule was 18 ± 1, 20 ± 1, 29 ± 2, 30 ± 2, 68 ± 5, 114 ± 8, 186 ± 10, 400 ± 20, and 447 ± 24. Maximum percentage of spermatogonia was 72 ± 2% at 6 wk. Primary and secondary spermatocytes were first observed at 10 and 12 wk of age, respectively, whereas spermatids were first apparent at 4 months. Sperm were first identified at 5 months, with more present at 6 months. We concluded that the optimal age for retrieving testicular germ cells in guinea fowl was no later than 8 wk, as that represented the age when seminiferous tubules were dominated by spermatogonia. The study was supported by the Russian Science Foundation (Project no.16-16-04104).

201 Spermatogonia Transplantation in the Chicken

2018 ◽  
Vol 30 (1) ◽  
pp. 241
Author(s):  
A. N. Vetokh ◽  
N. A. Volkova ◽  
T. O. Kotova ◽  
E. N. Antonova ◽  
A. V. Dotsev ◽  
...  
Keyword(s):  
Cell Population ◽  
Sertoli Cells ◽  
Recombinant Dna ◽  
Genetic Material ◽  
Live Weight ◽  
Seminiferous Tubules ◽  
Control Group ◽  
Spermatogenic Cells ◽  
Donor Cells ◽  
Male Chicken

Spermatogonia are the precursors of male germ cells. They are a valuable genetic material for the production of transgenic poultry. This technology includes isolation of the spermatogonia from male donor’s testes, transformation, and transplantation of donor cells into the sterilized recipient’s testes. The transplanted spermatogonia subsequently differentiate into male sex cells (sperm). The aim of this study was to optimize the individual stages of donor spermatogonia transplantation into the recipient’s testes to increase the effectiveness of spermatogenesis recovery. In the first stage, the spermatogenesis in male chicken was examined to determine the optimal age for isolation of spermatogonia from testes. Histological examinations of male chicken testes (n = 80 birds) were done for 8 age categories, from 1 week to 3 months. It was found that under the age of 4 weeks, the cell population in the seminiferous tubules of male chickens was represented mainly by Sertoli cells and spermatogonia. Maximum percentage of spermatogonia was 69 ± 3% at 4 weeks. At the next stage, a culture of spermatogonia was obtained. Testes of 3-week-old male chickens were used. Separation of the spermatogonia from other types of cells was based on a differential adhesive capacity. The maximum homogeneity of the cell population was established by transfer (3 times) of the supernatant containing unattached cells after 24 h of cultivation into a new culture dish for further cultivation. The cell population is represented mainly by the spermatogonia (89 ± 3%). The lentiviral transduction (pHAGE vector, ZsGreen under CMV promotor) was used to transform the resulting culture of the spermatogonia. The efficiency of spermatogonia infection with lentiviral particles (TU/mL = 2.5 × 108) was 65 ± 2%. After transformation, spermatogonia were introduced into the testes of busulfan-sterilized recipients. The optimal concentration of busulfan treatment after series of experiments from 40 to 100 mg/kg was determined. The effective dose for the removal of own spermatogenic cells was revealed at a concentration of 80 mg/kg of live weight. With complete elimination of other types of spermatogenic cells, the number of Sertoli cells and spermatogonia in the testicle tubules decreased by 39 ± 2% and 98 ± 1%, respectively, compared with the control group. The efficiency of spermatogenesis recovery was assessed based on sperm analysis that was obtained from male recipients (n = 5 birds) 4 months after the introduction of donor cells using PCR. The presence of recombinant DNA (ZsGreen) in recipients’ sperm was shown. Thus, our results indicate the prospect of using spermatogonia as a genetic material for the production of transgenic poultry. Study was supported by the Russian Science Foundation (Project no.16-16-10059).

scholarly journals PSXIII-5 The age dynamics of the spermatogenic cells development in the roosters’ testes

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 373-373
Author(s):  
Anastasia N Vetokh ◽  
Natalia A Volkova ◽  
Evgeniya K Tomgorova ◽  
Ludmila A Volkova ◽  
Natalia A Zinovieva
Keyword(s):  
Germ Cells ◽  
Sertoli Cells ◽  
Genetic Material ◽  
Cell Types ◽  
Seminiferous Tubules ◽  
Sperm Cells ◽  
Spermatogenic Cells ◽  
Different Cell Types ◽  
Hematoxylin Eosin ◽  
Testis Tissue

Abstract The cells of the male gonads are considered as a valuable genetic material for the conservation of the gene pool of breeds and lines of agricultural birds, as well as the directed modification of the poultry genome. Mature germ cells – spermatozoa and their predecessors – spermatogonia, spermatocytes and spermatids can be used for these purposes. To obtain these types of cells, it is necessary to know the characteristics of their development (spermatogenesis). The dynamics of the development of certain spermatogenic cell types in the testicular tubules of different-aged roosters has been studied. Histological studies were performed on testes of roosters aged from 1 week to 6 months with an interval of 2 weeks. Samples of testis tissue were fixed in Bouin’s solution. Histological sections were stained with hematoxylin-eosin. Identification of different cell types (Sertoli, spermatogonia, spermatocytes, spermatids, sperm cells) was carried out according to their morphology. At the age of 1–6 weeks in the seminiferous tubule of roosters, the mainly presence of two cell types was noted: Sertoli cells and spermatogonia. From 7 weeks of age, spermatocytes were detected in the seminiferous tubules, in the 4 months - spermatids, in the 5.5 months - sperm cells. The number of Sertoli cells remained almost unchanged with age and was 21 ± 2. The percentage of these cells decreased with age from 71 ± 3 % to 5 ± 1 %. The percentage of spermatogonia also decreased with age from 75 ± 2 % to 7 ± 1 %. The number of spermatids and spermatozoa, on the contrary, increased to puberty (6 months) and reached 54 %. The study was supported by the RFBR within Project no.18-29-07079.

scholarly journals Induced pluripotent stem cells-derived neurons from patients with Friedreich ataxia exhibit differential sensitivity to resveratrol and nicotinamide

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Pauline Georges ◽  
Maria-Gabriela Boza-Moran ◽  
Jacqueline Gide ◽  
Georges Arielle Pêche ◽  
Benjamin Forêt ◽  
...  
Keyword(s):  
Stem Cells ◽  
Drug Discovery ◽  
Pluripotent Stem Cells ◽  
Friedreich Ataxia ◽  
Cell Types ◽  
Differential Sensitivity ◽  
Human Ipscs ◽  
Induced Pluripotent ◽  
Species Specific

Abstract Translation of pharmacological results from in vitro cell testing to clinical trials is challenging. One of the causes that may underlie these discrepant results is the lack of the phenotypic or species-specific relevance of the tested cells; today, this lack of relevance may be reduced by relying on cells differentiated from human pluripotent stem cells. To analyse the benefits provided by this approach, we chose to focus on Friedreich ataxia, a neurodegenerative condition for which the recent clinical testing of two compounds was not successful. These compounds, namely, resveratrol and nicotinamide, were selected because they had been shown to stimulate the expression of frataxin in fibroblasts and lymphoblastoid cells. Our results indicated that these compounds failed to do so in iPSC-derived neurons generated from two patients with Friedreich ataxia. By comparing the effects of both molecules on different cell types that may be considered to be non-relevant for the disease, such as fibroblasts, or more relevant to the disease, such as neurons differentiated from iPSCs, a differential response was observed; this response suggests the importance of developing more predictive in vitro systems for drug discovery. Our results demonstrate the value of utilizing human iPSCs early in drug discovery to improve translational predictability.

scholarly journals Genome-wide identification and characterisation of HOT regions in the human genome

2016 ◽  
Author(s):  
Hao Li ◽  
Feng Liu ◽  
Chao Ren ◽  
Xiaochen Bo ◽  
Wenjie Shu
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Embryonic Stem Cells ◽  
Epigenetic Regulation ◽  
Human Cell ◽  
Embryonic Stem ◽  
Cell Types ◽  
Genome Wide ◽  
Development And Differentiation ◽  
Epigenetic Analysis

AbstractHOT (high-occupancy target) regions, which are bound by a surprisingly large number of transcription factors, are considered to be among the most intriguing findings of recent years. An improved understanding of the roles that HOT regions play in biology would be afforded by knowing the constellation of factors that constitute these domains and by identifying HOT regions across the spectrum of human cell types. We characterised and validated HOT regions in embryonic stem cells (ESCs) and produced a catalogue of HOT regions in a broad range of human cell types. We found that HOT regions are associated with genes that control and define the developmental processes of the respective cell and tissue types. We also showed evidence of the developmental persistence of HOT regions at primitive enhancers and demonstrate unique signatures of HOT regions that distinguish them from typical enhancers and super-enhancers. Finally, we performed an epigenetic analysis to reveal the dynamic epigenetic regulation of HOT regions upon H1 differentiation. Taken together, our results provide a resource for the functional exploration of HOT regions and extend our understanding of the key roles of HOT regions in development and differentiation.

scholarly journals Expression of Promyelocytic Leukemia (PML) Protein in the Mouse Developing Spermatogonia

2020 ◽  
Author(s):  
Amin Tavassoli ◽  
Hesam DEHGHANI
Keyword(s):  
Stem Cells ◽  
Immunohistochemical Analysis ◽  
Cell Types ◽  
Spermatogonial Stem Cells ◽  
Promyelocytic Leukemia ◽  
Nuclear Bodies ◽  
Seminiferous Tubules ◽  
Open Chromatin ◽  
Pml Nuclear Bodies ◽  
Oct4 Protein

Abstract Background: Promyelocytic leukemia (PML) as the main protein of PML nuclear bodies regulates various physiological processes such as transcription, DNA repair, apoptosis, senescence, and several signaling pathways in different cell types. It is well known that the PML protein is involved in the regulation of stem cell properties by maintaining an open chromatin conformation for the regulatory regions of the Oct4 gene. However, there is no experimental evidence for the presence and function of PML protein in the testis tissue. Results: In this study, we show the presence of PML protein in the developing mouse testis and its co-expression with the OCT4 protein. Immunohistochemical analysis of testis mirror sections shows that PML is co-expressed with the OCT4 protein in the outermost cellular layer of seminiferous tubules, where the spermatogonial stem cells are located. Conclusions: Our findings suggest that the PML protein might be involved in the stemness of spermatogonial stem cells at different stages of its development, even before earning the ability to produce mature sperm.

142 Isolation of the Quail Spermatogonia

2018 ◽  
Vol 30 (1) ◽  
pp. 211
Author(s):  
E. R. Mennibaeva ◽  
N. A. Volkova ◽  
E. K. Tomgorova ◽  
L. A. Volkova ◽  
V. A. Bagirov ◽  
...  
Keyword(s):  
Growth Factor ◽  
Sertoli Cells ◽  
Recombinant Dna ◽  
Initial Study ◽  
Seminiferous Tubules ◽  
Target Cells ◽  
Spermatogenic Cells ◽  
Russian Science ◽  
Epidermal Growth ◽  
The Individual

Spermatogonia are testicular stem cells, the precursors of male sex cells. They are target cells for introduction of recombinant DNA and suitable for creation of cryobanks to preserve biological materials. The aim of our research was to optimize the individual stages culturing quail spermatogonia. In an initial study, dynamics of change in the composition of spermatogenic cells in the seminiferous tubules were assessed histologically, at weekly intervals from 1 week to 1.5 months of age. Thereafter, spermatogonia were isolated from quail testes. Disaggregation of the testis tissue was carried out by consecutive enzymatic treatment in 0.25% trypsin and 0.1% collagenase solution. Purification of spermatogonia from other types of spermatogenic cells was conducted by separation of the cells by adhesion. The duration and conditions of cultivation of spermatogenic cells were selected experimentally. Cultivation of spermatogonia was performed on feeder layers, including quail primary Sertoli cells, STO cell line, and transplanted porcine Sertoli cells. Growth medium for culturing spermatogonia was DMEM HG medium supplemented with 5% FCS, 2 mM α-glutamine, MEM (10 μL mL−1), antibiotic (100×), insulin-transferrin-selenium (ITS, 10 μL mL−1), 2-mercaptoethanol (5 × 10−5 M), albumin (5 mg mL−1), epidermal growth factor (EGF, 20 ng mL−1), basic fibroblast growth factor (bFGF, 10 ng mL−1), and leukemia inhibitory factor (LIF, 2 ng mL−1). For identification of spermatogonia colonies, SSEA-1 antibodies were used. The maximum number of spermatogonia in seminiferous tubules of quail occurred at 3 weeks of age; there were mainly spermatogonia and Sertoli cells at this time. The percentage of spermatogonia from the total number of spermatogenic cells in the seminiferous tubule reached 76 ± 2%. In view of this, spermatogonia were isolated from the testes of 2-week-old quail. Spermatogenic cells were cultured for 24 h, after which the supernatant with unattached cells, mainly spermatogonia, was transferred to a new dish and cultured. Maximum homogeneity of the cell population was detected by dividing the cells by 3-fold transfer of the cell supernatant at an interval of 24 h; the proportion of spermatogonia in the suspension reached 88%. Quail Sertoli cells were the optimal feeder layer for cultivation of quail spermatogonia. Formation of spermatogonia colonies was observed on Day 5 to 7 of cultures, and their identity confirmed by immunohistochemical staining for SSEA-1. The study was supported by the Russian Science Foundation within Project no.16-16-04104.

209 Rho signaling-directed regulation of YAP/TAZ in parthenogenetic stem cells

2020 ◽  
Vol 32 (2) ◽  
pp. 233
Author(s):  
G. Pennarossa ◽  
S. Arcuri ◽  
F. Gandolfi ◽  
T. Brevini
Keyword(s):  
Stem Cells ◽  
Molecular Mechanisms ◽  
Rho Gtpase ◽  
Hippo Pathway ◽  
Genetic Material ◽  
Embryonic Stem ◽  
Cell Types ◽  
Embryonic Cell ◽  
And Migration ◽  
The Hippo Pathway

Mammalian oocyte maturation is characterised by asymmetric meiotic division that is regulated by specific cytoskeleton organisation. Similarly, during early embryonic divisions, one of the most important steps is the establishment of polarity that allows cells to adopt distinct developmental fates. All of these events are driven by dynamic changes in actin filaments. It has been demonstrated recently that the Rho signalling pathway plays a key role in the organisation and rearrangement of actin-containing structures, regulating cell polarity and migration. In addition, beside its effect on cell cytoskeleton, Rho directly interacts with the Hippo pathway, influencing both embryonic cell proliferation and differentiation. Because both Rho and Hippo are expressed by the oocyte and maternally inherited (Zhang et al. 2014 Cell Cycle 13, 3390-3403, https://doi.org/10.4161/15384101.2014.952967; Menchero et al. 2017 Dev. Dyn. 246, 245-261, https://doi.org/10.1002/dvdy.24471), we investigated their regulation in parthenogenetic embryonic stem cells (ParthESC) that possess exclusively maternal genetic material, and compared the results with biparental ESCs. Previous results obtained by whole-transcriptome analysis revealed the presence of several differentially expressed genes involved in the Rho pathway and showed no differences for most of the Hippo signalling genes. To better elucidate the molecular mechanisms involved, in the present study, we dissected the expression pattern of the Rho and Hippo regulatory genes in human biparental ESCs and ParthESC. Experiments were performed on 4 biparental ESC and 4 ParthESC lines using cells between passages 5 to 25. The results showed significantly increased transcription of the Rho GTPase family genes (RHOA, RHOB, and RHOC) in ParthESC compared with biparental ESCs. Consistent with this, 12 of 17 Rho activators were significantly upregulated, whereas 8 of 11 Rho inhibitors were significantly decreased in ParthESC. Furthermore, monoparental cells displayed significantly higher expression levels of YAP and TAZ, whereas the upstream genes involved in the Hippo pathway (LATS1/2, MOB1, MST1/2, NF2) were comparable in the two cell types. Interestingly, a significantly higher total YAP protein content was detected in ParthESC, whereas the quantity of the phosphorylated form was comparable in the two cell types. This accounts for the observed upregulation of Rho genes, which stimulate the assembly of contractile actin stress fibres, inhibiting LATS1/2 phosphorylation and preventing subsequent phosphorylation of YAP/TAZ (Yu and Guan 2013 Genes Dev. 27, 355-371; https://doi.org/10.1101/gad.210773.112). Altogether, our results suggest that the Rho pathway may regulate YAP/TAZ behaviour via a LATS/MST/NF2-independent process in ParthESC, similarly to a previous report in oocytes (Posfai and Rossant 2016 Cell Res. 26, 393-394; https://doi.org/10.1038/cr.2016). Although further clarifications are needed, we hypothesise that the regulatory mechanisms detected in ParthESC may be related to their strictly maternal origin, with a possible impact on their plasticity and potency. This study was supported by Carraresi Foundation. Authors are members of the COST Actions CA16119.

137 Recovery of quail spermatogenesis by donor spermatogonia transplantation

2020 ◽  
Vol 32 (2) ◽  
pp. 195
Author(s):  
A. N. Vetokh ◽  
E. K. Tomgorova ◽  
L. A. Volkova ◽  
N. A. Volkova ◽  
N. A. Zinovieva
Keyword(s):  
Germ Cells ◽  
Recombinant Dna ◽  
Seminiferous Tubules ◽  
Spermatogenic Cells ◽  
Optimal Parameters ◽  
Histological Studies ◽  
Sperm Dna ◽  
Donor Cells ◽  
Reported Study ◽  
Series Of Experiments

Spermatogonia, the stem cell precursors of male germ cells, are used as convenient biological material for the preservation of genetic resources (cryobanks) and the introduction of recombinant DNA (transgenesis). Donor spermatogonia subsequently differentiate into mature germ cells (spermatozoa), which are used to produce offspring. Our laboratory is investigating methods to improve the efficiency of spermatogonial germ cell transplantation in quail. The objectives of this study were to (1) determine the optimal age for spermatogonia isolation from the testes of donor male quail and (2) identify the most appropriate concentration of busulfan for treatment of recipient quail testes. Statistical analysis was performed using SPSS ver. 15.0 (IBM Corp.; analysis of variance test). In order to determine the optimal age for spermatogonia isolation, testes from male quail at 1-4 weeks of age were isolated and histological studies were performed on a population of spermatogenic cells in the seminiferous tubules. Histological studies of quail testes isolated at different ages showed that the optimal age for obtaining a culture of spermatogonia is a period from 1-2 weeks of age. During this period, spermatogenic cells were represented mainly by spermatogonia (P<0.01). Therefore, testes of 1-week-old quails were used to obtain a culture of spermatogonia. The resulting cell culture consisted mainly of spermatogonia (85%) with a small number of Sertoli cells. Next, a series of experiments introducing busulfan into quail testes was carried out using concentrations from 10-150mg kg−1 of liveweight. Experiments showed that an effective dose to remove the recipient male's own spermatogenic cells was a concentration of 100mg kg−1 of liveweight (P<0.05). Finally, using the optimal parameters described above, spermatogonia cultures were obtained and introduced into the testes of quail recipients (n=6), following administration of busulfan for 2-3 weeks before donor spermatogonia were introduced. The effectiveness of spermatogenesis recovery was assessed based on the analysis of sperm from quail recipients at 3 months after the injection of donor cells. The presence of donor germ cells in the testes of quail recipient drakes was confirmed by microsatellite analysis of DNA isolated from the blood and sperm of recipients as well as the donor cells (spermatogonia). The microsatellite profiles of the blood and sperm DNA in quail recipient males were different, which confirms the restoration of spermatogenesis in the studied recipients due to the development of donor germ cells. The reported study was funded by RFBR, project number 18-29-07079.

In vitro development of nuclear transfer embryos derived from porcine embryonic germ cells and their descendent neural precursor cells

Zygote ◽  
2011 ◽  
Vol 20 (1) ◽  
pp. 9-15 ◽  
Author(s):  
Susa Shin ◽  
Kwang Sung Ahn ◽  
Seong-Jun Choi ◽  
Soon Young Heo ◽  
Hosup Shim
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
Nuclear Transfer ◽  
Primordial Germ Cells ◽  
Somatic Cells ◽  
Cell Types ◽  
Neural Precursor ◽  
Blastocyst Development ◽  
Donor Cells

SummaryUndifferentiated stem cells may support a greater development of cloned embryos compared with differentiated cell types due to their ease of reprogramming during the nuclear transfer (NT) process. Hence, stem cells may be more suitable as nuclear donor cells for NT procedures than are somatic cells. Embryonic germ (EG) cells are undifferentiated stem cells that are isolated from cultured primordial germ cells (PGC) and can differentiate into several cell types. In this study, the in vitro development of NT embryos using porcine EG cells and their derivative neural precursor (NP) cells was investigated, thus eliminating any variation in genetic differences. The rates of fusion did not differ between NT embryos from EG and NP cells; however, the rate of cleavage in NT embryos derived from EG cells was significantly higher (p < 0.05) than that from NP cells (141/247 [57.1%] vs. 105/228 [46.1%]). Similarly, the rate of blastocyst development was significantly higher (P < 0.05) in NT using EG cells than the rate using NP cells (43/247 [17.4%] vs. 18/228 [7.9%]). The results obtained from the present study in pigs demonstrate a reduced capability for nuclear donor cells to be reprogrammed following the differentiation of porcine EG cells. Undifferentiated EG cells may be more amenable to reprogramming after reconstruction compared with differentiated somatic cells.

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