Production of somatic and germline chimeras in the chicken by transfer of early blastodermal cells

Development ◽  
1990 ◽  
Vol 108 (1) ◽  
pp. 185-189 ◽  
Author(s):  
J.N. Petitte ◽  
M.E. Clark ◽  
G. Liu ◽  
A.M. Verrinder Gibbins ◽  
R.J. Etches
Keyword(s):  
Inbred Line ◽  
Cell Lineage ◽  
Donor Cell ◽  
Black Pigment ◽  
Dominant Allele ◽  
White Leghorn ◽  
Sperm Dna ◽  
Donor Cells ◽  
White Leghorns ◽  
Blastodermal Cells

Cells were isolated from stage X embryos of a line of Barred Plymouth Rock chickens (that have black pigment in their feathers due to the recessive allele at the I locus) and injected into the subgerminal cavity of embryos from an inbred line of Dwarf White Leghorns (that have white feathers due to the dominant allele at the I locus). Of 53 Dwarf White Leghorn embryos that were injected with Barred Plymouth Rock blastodermal cells, 6 (11.3%) were phenotypically chimeric with respect to feather colour and one (a male) survived to hatching. The distribution of black feathers in the recipients was variable and not limited to a particular region although, in all but one case, the donor cell lineage was evident in the head. The male somatic chimera was mated to several Barred Plymouth Rock hens to determine the extent to which donor cells had been incorporated into his testes. Of 719 chicks hatched from these matings, 2 were phenotypically Barred Plymouth Rocks demonstrating that cells capable of incorporation into the germline had been transferred. Fingerprints of the blood and sperm DNA from the germline chimera indicated that both of these tissues were different from those of the inbred line of Dwarf White Leghorns. Bands that were present in fingerprints of blood DNA from the chimera and not present in those of the Dwarf White Leghorns were observed in those of the Barred Plymouth Rocks.(ABSTRACT TRUNCATED AT 250 WORDS)

Germline chimeric chickens from dispersed donor blastodermal cells and compromised recipient embryos

Development ◽  
1993 ◽  
Vol 117 (2) ◽  
pp. 669-675 ◽  
Author(s):  
R.S. Carsience ◽  
M.E. Clark ◽  
A.M. Verrinder Gibbins ◽  
R.J. Etches
Keyword(s):  
Gamma Radiation ◽  
Recipient Cell ◽  
Recessive Allele ◽  
Dominant Allele ◽  
White Leghorn ◽  
60Co Source ◽  
Donor Cells ◽  
Black Pigmentation ◽  
Dispersed Cells ◽  
Blastodermal Cells

Stage-X blastoderms, within intact eggs from White Leghorn hens, were exposed to 500–700 rads of gamma radiation from a 60Co source prior to injection, into the subgerminal cavity, of approximately 100 or 200–400 dispersed cells from stage-X blastoderms isolated from eggs laid by Barred Plymouth Rock hens. Embryos developing past day 14 of incubation and hatched chicks were assessed for donor and recipient cell contribution to the melanocyte population through examination of black and yellow down pigmentation, respectively (Barred Plymouth Rocks have a recessive allele at the I locus while the White Leghorns have a dominant allele at the I locus). Of the 809 embryos injected with approximately 100 cells, 192 developed past day 14 and black pigmentation, indicating somatic chimerism, was observed on 118 of the 192 (58%) embryos and chicks. Of the 296 embryos injected with 200–400 donor cells, 86 developed past day 14 of incubation. Somatic chimerism was observed on 55 of the 86 (64%) embryos and chicks. To test for germline chimerism, birds surviving to maturity were mated to Barred Plymouth Rocks. Five somatically chimeric females were produced when approximately 100 cells were injected, and one was a germline chimera. Six somatic female chimeras were produced following the injection of 200–400 cells, three of which proved to be germline chimeras by the presence of Barred Rock chicks among their offspring. Two of the nine males produced by injecting approximately 100 cells were germline chimeras.(ABSTRACT TRUNCATED AT 250 WORDS)

46 RECLONING USING TRANSGENIC FETAL FIBROBLASTS AS NUCLEI DONORS INCREASES DEVELOPMENTAL POTENTIAL OF RECONSTRUCTED EMBRYOS IN CATTLE

2010 ◽  
Vol 22 (1) ◽  
pp. 180
Author(s):  
F. F. Bressan ◽  
M. S. Miranda ◽  
F. Perecin ◽  
T. H. C. De Bem ◽  
M. Bajgelman ◽  
...  
Keyword(s):  
Genetically Modified ◽  
Cell Lineage ◽  
Donor Cell ◽  
Pregnancy Rates ◽  
Control Group ◽  
Specific Cell ◽  
Nuclear Reprogramming ◽  
Developmental Potential ◽  
Donor Cells ◽  
Fetal Fibroblasts

Genetically modified animals have numerous applications ranging from basic research to agriculture production. Cloning by nuclear transfer (NT) has made possible the production of transgenic animals using previously genetically modified cell lineages. Gene expression studies and adequate selection of the nuclei donor cell for NT guarantees the presence of the gene construction in the offspring and the absence of deleterious mutations caused by the random insertion of transgenes in functional areas of the genome. Embryonic development after NT requires a change in the transcriptome of the donor cell from a somatic to an embryonic pattern, causing cloning efficiencies to be low because of incomplete or defective nuclear reprogramming. Therefore, the establishment of methodologies able to increase cloning success is highly desirable. The experiment was designed to test if recloning of transgenic fetal fibroblasts increases cloned blastocyst production and the pregnancy rates of transgenic cloned embryos produced by NT. This study compared the developmental potential of cloned embryos reconstructed with fetal fibroblasts genetically modified by lentivirus random integration (control group) expressing the green fluorescent protein gene (eGFP), with a transgenic fetal fibroblast cell line established from a 30-day transgenic pregnancy (recloning group). Fusion, cleavage (72 h post-activation, hpa), blastocyst production (168 hpa), and 30-day pregnancy rates were analyzed. A total of 1213 embryos were reconstructed; 884 (10 replicates) with random transgene insertion fibroblasts and 329 (4 replicates) with cells derived from the cloned fetus. Results were analyzed by chi-square test at 5% significance. No difference was observed (P > 0.05) between control and recloned groups regarding fusion rate (n = 550, 62.22% and n = 189, 57.25%; respectively) or cleavage rate (n = 383, 69.45% and n = 132, 69.84%, respectively). The recloned group, however, showed a higher blastocyst development rate (P < 0.01) compared with the control group (n = 51, 26.98%, and n = 79, 14.36%, respectively) and a higher 30-day pregnancy rate (n = 6, 15.38% and n = 3, 5.56%, respectively). In conclusion, recloning of transgenic fibroblasts from a successfully established pregnancy augments the efficiency in the production of embryos and pregnancy establishment compared with the control group. We speculate that a second round of NT enhances the nuclear reprogramming of donor cells, and moreover, the use of a transgenic cell lineage already proven to be successfully reprogrammed may indicate that transgene integration is not deleterious in that specific cell lineage, resulting in a good source of donor cells to be used in order to produce a homogeneous bioreactor herd. Financial support: FAPESP, Brazil.

scholarly journals 1DEVELOPMENT OF BOVINE NUCLEAR TRANSFER EMBRYOS CLONED FROM FOLLICULAR DONOR CELLS IN SEQUENTIAL STAGES OF DIFFERENTIATION

2004 ◽  
Vol 16 (2) ◽  
pp. 123 ◽  
Author(s):  
C.A. Batchelder ◽  
K.A. Hoffert ◽  
M. Bertolini ◽  
A.L. Moyer ◽  
G.B. Anderson
Keyword(s):  
Comparative Studies ◽  
Nuclear Transfer ◽  
Es Cells ◽  
Block Design ◽  
Cell Lineage ◽  
Donor Cell ◽  
Cell Types ◽  
Efficient Production ◽  
Differentiated Cells ◽  
Donor Cells

Efficient production of cloned embryos and live offspring is dependent on the ability of the nuclear-donor cell to be reprogrammed to direct normal conceptus development. Results of comparative studies indicate that embryonic and fetal cells are more successful candidates for nuclear transfer (NT) than terminally differentiated cells. Comparison of donor-cell efficiency is difficult to interpret across laboratories and from donor animals of varying genetic backgrounds and tissues of origin. This study was undertaken to determine the effect of the stage of differentiation of adult somatic donor cells derived from one-cell lineage of an individual donor animal. The follicular cell lineage including preantral follicle (PAFC), cumulus (CC), granulosa (GC), and luteal cells (LC) was chosen as several cell types in the lineage have previously proven successful for NT. Donor cell cultures were established from a 3-year-old Holstein cow. Embryos were reconstructed using confluent, early passage cultures. For each replicate, embryos were produced from two donor cell types in randomized block design (14 trials). Viable embryos were transferred to recipient females after 7 days of in vitro culture. Pregnancy and fetal viability were monitored weekly by ultrasonography from Days 30–100 of gestation and by rectal palpation thereafter. Embryos reconstructed from PAFC were less likely to develop to the blastocyst stage on Day 7 than embryos derived from CC or LC (Table 1, P&lt;0.05). Pregnancy rates at Day 30 were similar across donor cell types. A greater proportion of PAFC embryos were viable at Day 60 of gestation than embryos derived from CC and GC (P&lt;0.05). To date, normal cloned calves have been delivered at term from CC and GC, and two pregnancies (n=3 fetuses) are ongoing from PAFC. The fetus cloned from LC, the terminally differentiated cell type in the lineage, was spontaneously aborted at day 211 with congenital abnormalities. Results from comparative studies of development of mouse embryos cloned from embryonic stem (ES) cells v. somatic donor cells indicate that ES-derived clones are less efficient in blastocyst formation, but survival to term is greater (Humpherys D et al., 2002 PNAS 99, 12889–12894). Likewise, our results in cattle suggest that PAFC, the least differentiated cells in the lineage, result in fewer cloned blastocysts, but blastocysts that do develop are more likely to progress through implantation and into later stages of pregnancy. Table 1. Development of NT embryos reconstructed from follicular donor cells at sequential stages of differentiation

scholarly journals Dynamic integration of enteric neural stem cells in ex vivo organotypic colon cultures

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Georgina Navoly ◽  
Conor J. McCann
Keyword(s):  
Stem Cells ◽  
Neural Stem Cells ◽  
Organotypic Culture ◽  
Ex Vivo ◽  
Donor Cell ◽  
Colonic Tissue ◽  
Cell Therapies ◽  
Enteric Ganglia ◽  
Donor Cells

AbstractEnteric neural stem cells (ENSC) have been identified as a possible treatment for enteric neuropathies. After in vivo transplantation, ENSC and their derivatives have been shown to engraft within colonic tissue, migrate and populate endogenous ganglia, and functionally integrate with the enteric nervous system. However, the mechanisms underlying the integration of donor ENSC, in recipient tissues, remain unclear. Therefore, we aimed to examine ENSC integration using an adapted ex vivo organotypic culture system. Donor ENSC were obtained from Wnt1cre/+;R26RYFP/YFP mice allowing specific labelling, selection and fate-mapping of cells. YFP+ neurospheres were transplanted to C57BL6/J (6–8-week-old) colonic tissue and maintained in organotypic culture for up to 21 days. We analysed and quantified donor cell integration within recipient tissues at 7, 14 and 21 days, along with assessing the structural and molecular consequences of ENSC integration. We found that organotypically cultured tissues were well preserved up to 21-days in ex vivo culture, which allowed for assessment of donor cell integration after transplantation. Donor ENSC-derived cells integrated across the colonic wall in a dynamic fashion, across a three-week period. Following transplantation, donor cells displayed two integrative patterns; longitudinal migration and medial invasion which allowed donor cells to populate colonic tissue. Moreover, significant remodelling of the intestinal ECM and musculature occurred upon transplantation, to facilitate donor cell integration within endogenous enteric ganglia. These results provide critical evidence on the timescale and mechanisms, which regulate donor ENSC integration, within recipient gut tissue, which are important considerations in the future clinical translation of stem cell therapies for enteric disease.

scholarly journals Rabbit somatic cell cloning: effects of donor cell type, histone acetylation status and chimeric embryo complementation

Reproduction ◽  
2007 ◽  
Vol 133 (1) ◽  
pp. 219-230 ◽  
Author(s):  
Feikun Yang ◽  
Ru Hao ◽  
Barbara Kessler ◽  
Gottfried Brem ◽  
Eckhard Wolf ◽  
...  
Keyword(s):  
Histone Acetylation ◽  
Nuclear Transfer ◽  
Donor Cell ◽  
Epigenetic Mark ◽  
Cell Type ◽  
Developmental Potential ◽  
Acetylation Status ◽  
Donor Cells ◽  
Donor Cell Type

The epigenetic status of a donor nucleus has an important effect on the developmental potential of embryos produced by somatic cell nuclear transfer (SCNT). In this study, we transferred cultured rabbit cumulus cells (RCC) and fetal fibroblasts (RFF) from genetically marked rabbits (Alicia/Basilea) into metaphase II oocytes and analyzed the levels of histone H3-lysine 9-lysine 14 acetylation (acH3K9/14) in donor cells and cloned embryos. We also assessed the correlation between the histone acetylation status of donor cells and cloned embryos and their developmental potential. To test whether alteration of the histone acetylation status affects development of cloned embryos, we treated donor cells with sodium butyrate (NaBu), a histone deacetylase inhibitor. Further, we tried to improve cloning efficiency by chimeric complementation of cloned embryos with blastomeres fromin vivofertilized or parthenogenetic embryos. The levels of acH3K9/14 were higher in RCCs than in RFFs (P<0.05). Although the type of donor cells did not affect development to blastocyst, after transfer into recipients, RCC cloned embryos induced a higher initial pregnancy rate as compared to RFF cloned embryos (40 vs 20%). However, almost all pregnancies with either type of cloned embryos were lost by the middle of gestation and only one fully developed, live RCC-derived rabbit was obtained. Treatment of RFFs with NaBu significantly increased the level of acH3K9/14 and the proportion of nuclear transfer embryos developing to blastocyst (49 vs 33% with non-treated RFF,P<0.05). The distribution of acH3K9/14 in either group of cloned embryos did not resemble that inin vivofertilized embryos suggesting that reprogramming of this epigenetic mark is aberrant in cloned rabbit embryos and cannot be corrected by treatment of donor cells with NaBu. Aggregation of embryos cloned from NaBu-treated RFFs with blastomeres fromin vivoderived embryos improved development to blastocyst, but no cloned offspring were obtained. Two live cloned rabbits were produced from this donor cell type only after aggregation of cloned embryos with a parthenogenetic blastomere. Our study demonstrates that the levels of histone acetylation in donor cells and cloned embryos correlate with their developmental potential and may be a useful epigenetic mark to predict efficiency of SCNT in rabbits.

The inheritance of shank colour in single comb White Leghorn and Australorp fowls

1951 ◽  
Vol 2 (4) ◽  
pp. 494
Author(s):  
F Skaller ◽  
GW Grigg
Keyword(s):  
White Leghorn ◽  
Genetical Analysis ◽  
Different Types ◽  
White Leghorns ◽  
Single Comb ◽  
Mode Of Inheritance

Evidence has been presented in the literature on the mode of inheritance of shank colour in various breeds of chickens, but little has hitherto been known about the same aspect in an Australian breed, the Black Australorp. Observations on the incidence of shank colour in different types of crosses between yellow-shanked White Leghorns and black-shanked Australorps are reported in this paper. According to the observations made by other workers on White Leghorn and other breeds, a hypothesis is advanced regarding the genes involved in determining shank colour in the White Leghorn and Australorp. A genetical analysis of the observations made by the authors on 847 single crosses, 71 backcrosses, and 31 zig-zag crosses is presented and shows close statistical agreement with the hypothesis. A genetical formula, including four autosomal and two sex-linked genes, is suggested which would explain the mode of inheritance of shank colour in White Leghorns and Australorps.

scholarly journals Manipulating the Mitochondrial Genome To Enhance Cattle Embryo Development

2017 ◽  
Vol 7 (7) ◽  
pp. 2065-2080 ◽  
Author(s):  
Kanokwan Srirattana ◽  
Justin C St. John
Keyword(s):  
Embryo Development ◽  
Nuclear Transfer ◽  
Bos Taurus ◽  
Trichostatin A ◽  
Donor Cell ◽  
Blastocyst Stage ◽  
Genetic Integrity ◽  
Rna Seq ◽  
Mtdna Copy Number ◽  
Donor Cells

Abstract The mixing of mitochondrial DNA (mtDNA) from the donor cell and the recipient oocyte in embryos and offspring derived from somatic cell nuclear transfer (SCNT) compromises genetic integrity and affects embryo development. We set out to generate SCNT embryos that inherited their mtDNA from the recipient oocyte only, as is the case following natural conception. While SCNT blastocysts produced from Holstein (Bos taurus) fibroblasts were depleted of their mtDNA, and oocytes derived from Angus (Bos taurus) cattle possessed oocyte mtDNA only, the coexistence of donor cell and oocyte mtDNA resulted in blastocysts derived from nondepleted cells. Moreover, the use of the reprogramming agent, Trichostatin A (TSA), further improved the development of embryos derived from depleted cells. RNA-seq analysis highlighted 35 differentially expressed genes from the comparison between blastocysts generated from nondepleted cells and blastocysts from depleted cells, both in the presence of TSA. The only differences between these two sets of embryos were the presence of donor cell mtDNA, and a significantly higher mtDNA copy number for embryos derived from nondepleted cells. Furthermore, the use of TSA on embryos derived from depleted cells positively modulated the expression of CLDN8, TMEM38A, and FREM1, which affect embryonic development. In conclusion, SCNT embryos produced by mtDNA depleted donor cells have the same potential to develop to the blastocyst stage without the presumed damaging effect resulting from the mixture of donor and recipient mtDNA.

scholarly journals Purified murine hematopoietic stem cells function longer on nonirradiated W41/Wv than on +/+ irradiated stroma

Blood ◽  
1993 ◽  
Vol 81 (6) ◽  
pp. 1489-1496 ◽  
Author(s):  
F Vecchini ◽  
KD Patrene ◽  
SS Boggs
Keyword(s):  
Stem Cells ◽  
Wheat Germ ◽  
Donor Cell ◽  
Mouse Bone Marrow ◽  
Hematopoietic Stem ◽  
Cell Production ◽  
Donor Cells ◽  
Nonadherent Cell ◽  
Specific Increase

Abstract Mouse bone marrow (BM) was separated into low-density, lineage- negative, wheat germ agglutinin-positive (WGA+), Rhodamine-123 bright (Rhbright) or dim (Rhdim) cells to obtain populations that were highly enriched for committed progenitors (Rhbright cells) or for more primitive stem cells (Rhdim). When 2,500 Rhbright or Rhdim cells were seeded onto 6-week-old irradiated (20 Gy) long-term BM cultures (LTBMC), the nonadherent cell production from Rhbright cells was transient and ended after 5 weeks. Production from Rhdim cells did not begin until week 3, peaked at week 5, and ended at week 8, when the irradiated stroma seemed to fail. Termination of cell production from Rhdim cells did not occur in nonirradiated LTBMC from W41/Wv mice. During peak nonadherent cell production, 25% to 30% of the cells in the nonirradiated LTBMC from W41/Wv mice had donor cell markers. Two approaches were tested to try to enhance the proportion or number of donor cells. Addition of Origen-HGF at the time of seeding Rhdim cells caused a nonspecific increase in both host and donor cell production, but a specific increase in production of donor cells was obtained by seeding the cultures at 2 weeks rather than 6 weeks. Limiting dilution of Rhdim cells gave the same frequency of wells producing cells on both irradiated +/+ and nonirradiated W41/Wv or W/Wv cultures.

scholarly journals Xenotransplantation of Human Neural Progenitor Cells to the Subretinal Space of Nonimmunosuppressed Pigs

2011 ◽  
Vol 2011 ◽  
pp. 1-6 ◽  
Author(s):  
Karin Warfvinge ◽  
Philip H. Schwartz ◽  
Jens Folke Kiilgaard ◽  
Morten la Cour ◽  
Michael J. Young ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Mononuclear Cell ◽  
Neural Progenitor Cells ◽  
Retinal Vasculitis ◽  
Donor Cell ◽  
Neural Progenitor ◽  
Morphological Integration ◽  
Subretinal Space ◽  
Donor Cells ◽  
Human Neural Progenitor Cells

To investigate the feasibility of transplanting human neural progenitor cells (hNPCs) to the retina of nonimmunosuppressed pigs, cultured hNPCs were injected into the subretinal space of 5 adult pigs after laser burns were applied to promote donor cell integration. Postoperatively, the retinal vessels appeared normal without signs of exudation, bleeding, or subretinal elevation. Eyes were harvested at 10–28 days. H&E consistently showed mild retinal vasculitis, depigmentation of the RPE, and marked mononuclear cell infiltrate in the choroid adjacent to the site of transplantation. Human-specific antibodies revealed donor cells in the subretinal space at 10–13 days and smaller numbers within the retina on days 12 and 13, with evidence suggesting a limited degree of morphological integration; however, no cells remained at 4 weeks. The strong mononuclear cell reaction and loss of donor cells indicate that modulation of host immunity is likely necessary for prolonged xenograft survival in this model.

Utilization of heterosis in poultry

1958 ◽  
Vol 9 (4) ◽  
pp. 599 ◽  
Author(s):  
FHW Morley
Keyword(s):  
Body Weight ◽  
Genetic Variation ◽  
Genetic Variability ◽  
Sex Chromosome ◽  
Inbred Lines ◽  
White Leghorn ◽  
Egg Weight ◽  
Additive Genetic Variation ◽  
White Leghorns ◽  
Recessive Genes

Records were analysed of 500-day production, egg weight, 11-week and mature body weight, sex maturity, and broodiness of the crossbred progeny of inbred lines of Australorps mated to White Leghorns, and inbred lines of White Leghorns mated to Australorps. Clear differences between lines of both breeds mere found in most characters, indicating the presence of potentially useful amounts of additive genetic variation. Non-additive genetic variation was also found to be present in varying degrees in different characters. Because of the difficulties of developing and maintaining inbred stocks of poultry, and the importance of sex-linked characteristics in some commercial environments, a scheme is proposed which may enable heterosis to be exploited without the use of inbred material. The basis of this scheme is the combination of the White Leghorn sex chromosome, with varying proportions of Australorp and White Leghorn autosomes, in a new breed. Assuming that the heterosis observed in the F1 is due to elimination of certain biochemical blocks determined by recessive genes, the formation of the new breed should enable the methods of closed flock breeding to be used in material at a higher level of production, and likely to contain more genetic variability, than either parent breed.

Sign in / Sign up

Export Citation Format

Share Document