scholarly journals Feeder Cell Type Affects the Growth of In Vitro Cultured Bovine Trophoblast Cells

2017 ◽  
Vol 2017 ◽  
pp. 1-6 ◽  
Author(s):  
Islam M. Saadeldin ◽  
Ahmed Abdelfattah-Hassan ◽  
Ayman Abdel-Aziz Swelum
Keyword(s):  
Granulosa Cells ◽  
Current Approach ◽  
Embryonic Cells ◽  
Interferon Tau ◽  
Porcine Granulosa Cells ◽  
Trophectoderm Cells ◽  
Homeobox Protein ◽  
First Time

Trophectoderm cells are the foremost embryonic cells to differentiate with prospective stem-cell properties. In the current study, we aimed at improving the current approach for trophoblast culture by using granulosa cells as feeders. Porcine granulosa cells (PGCs) compared to the conventional mouse embryonic fibroblasts (MEFs) were used to grow trophectoderm cells from hatched bovine blastocysts. Isolated trophectoderm cells were monitored and displayed characteristic epithelial/cuboidal morphology. The isolated trophectoderm cells expressed mRNA of homeobox protein (CDX2), cytokeratin-8 (KRT8), and interferon tau (IFNT). The expression level was higher on PGCs compared to MEFs throughout the study. In addition, primary trophectoderm cell colonies grew faster on PGCs, with a doubling time of approximately 48 hrs, compared to MEFs. PGCs feeders produced a fair amount of 17β-estradiol and progesterone. We speculated that the supplementation of sex steroids and still-unknown factors during the trophoblasts coculture on PGCs have helped to have better trophectoderm cell’s growth than on MEFs. This is the first time to use PGCs as feeders to culture trophectoderm cells and it proved superior to MEFs. We propose PGCs as alternative feeders for long-term culture of bovine trophectoderm cells. This model will potentially benefit studies on the early trophoblast and embryonic development in bovines.

64 USING PORCINE GRANULOSA CELLS AS FEEDERS FOR PORCINE AND BOVINE TROPHECTODERM CELL CULTURE

2012 ◽  
Vol 24 (1) ◽  
pp. 144
Author(s):  
I. M. Saadeldin ◽  
A. Elsayed ◽  
J. T. Kang ◽  
S. J. Park ◽  
S. J. Kim ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Doubling Time ◽  
Good Alternative ◽  
Blastocyst Stage ◽  
Outer Side ◽  
Embryonic Cells ◽  
Reverse Transcription Pcr ◽  
Porcine Granulosa Cells ◽  
Trophectoderm Cells

The trophectoderm cells, arising from the outer side of the blastomere in the blastocyst stage, are the first differentiated embryonic cells with specific potential as stem cells. The physiology of trophectoderm cells has been studied; however, their functions still remain unclear, because the lack of definitive information of cell lineages. Here, we aimed to establish in culture different feeder-dependent trophectoderm cell lines from 9-day, preimplantation, in vitro-produced porcine and bovine embryos. We used 2 different feeders: porcine granulosa cells (PGC) and mouse embryonic fibroblasts (MEF). Both cells were mitotically inactivated by mitomycin-C and then cultured with a density of 5 × 104 mL–1 on 0.1% (wt/vol) gelatin coated 4-well dishes in DMEM-199 medium supplemented with 10% (vol/vol) fetal bovine serum (FBS), nonessential amino acids (NEAA), β-mercaptoethanol and nucleosides (Talbot et al. 2000 Biol. Reprod. 62, 235–247). Trophectoderm cells were observed by light microscopy and characterised by reverse transcription-PCR using specific primers for both species. Different feeders and trophectoderm cells growth rates were compared after trypsinization using a hemocytometer. Data were analysed using 1-way ANOVA. In results, trophectoderm cells display epithelial characteristics, cuboidal morphology and express mRNA of homebox protein CDX2, cytokeratin 8 (KRT8) and interferon (IFN) gamma or tau for porcine or bovine cells, respectively. Moreover, oestrogen receptor (ESR1) and progesterone receptor (PGR) were expressed in trophectoderm cells of both species. Porcine granulosa cells were highly proliferative with doubling time of 24 h when compared to MEF (P ≤ 0.5), easy to recover and provided a reasonable source of steroids, 17β-oestradiol (E2; 31.21 ± 3.1 ng mL–1) and progesterone (P4; 6.36 ± 0.4 ng mL–1). Moreover, trophectoderm cell colonies of both species that cultured on PGC grew faster, with a doubling time of 48 h when compared to those cultured on MEF (P ≤ 0.5). We speculate that the continuous supplement of steroids and other cytokines during the co-culture of trophoblasts with granulosa cells might help the trophectoderm cells growth more than that of MEF. Further investigations are required in this regard. In conclusion, porcine granulosa cells can be good alternative feeders to culture porcine and bovine trophectoderm. This research was supported by MKE (Grant # 10033839-2011-13) and IPET.

Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

1970 ◽  
Vol 19 (1) ◽  
pp. 89-99
Author(s):  
K. Choudhary ◽  
M. Singh ◽  
M. S. Rathore ◽  
N. S. Shekhawat
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Long Term Study ◽  
Continuous Application ◽  
First Time ◽  
Pasture Legume

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l  of 2,4-D and 0.5 mg/l  of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture  legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

scholarly journals Characterization of Postharvest Fungicide-Resistant Botrytis cinerea Isolates From Commercially Stored Apple Fruit

2017 ◽  
Vol 107 (3) ◽  
pp. 362-368 ◽  
Author(s):  
Wayne M. Jurick ◽  
Otilia Macarisin ◽  
Verneta L. Gaskins ◽  
Eunhee Park ◽  
Jiujiang Yu ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Gray Mold ◽  
Apple Fruit ◽  
Sublethal Dose ◽  
Long Term Storage ◽  
Pose Control ◽  
First Time

Botrytis cinerea causes gray mold and is an economically important postharvest pathogen of fruit, vegetables, and ornamentals. Fludioxonil-sensitive B. cinerea isolates were collected in 2011 and 2013 from commercial storage in Pennsylvania. Eight isolates had values for effective concentrations for inhibiting 50% of mycelial growth of 0.0004 to 0.0038 μg/ml for fludioxonil and were dual resistant to pyrimethanil and thiabendazole. Resistance was generated in vitro, following exposure to a sublethal dose of fludioxonil, in seven of eight dual-resistant B. cinerea isolates. Three vigorously growing B. cinerea isolates with multiresistance to postharvest fungicides were further characterized and found to be osmosensitive and retained resistance in the absence of selection pressure. A representative multiresistant B. cinerea strain caused decay on apple fruit treated with postharvest fungicides, which confirmed the in vitro results. The R632I mutation in the Mrr1 gene, associated with fludioxonil resistance in B. cinerea, was not detected in multipostharvest fungicide-resistant B. cinerea isolates, suggesting that the fungus may be using additional mechanisms to mediate resistance. Results from this study show for the first time that B. cinerea with dual resistance to pyrimethanil and thiabendazole can also rapidly develop resistance to fludioxonil, which may pose control challenges in the packinghouse environment and during long-term storage.

RGD-mediated adhesion of porcine granulosa cells modulates their differentiation response to FSH in vitro

1995 ◽  
Vol 83 (2-3) ◽  
pp. 169-177 ◽  
Author(s):  
Béatrice Goxe ◽  
Jacques E. Flechon ◽  
Solange Delasalle ◽  
Roland Salesse
Keyword(s):  
Granulosa Cells ◽  
Porcine Granulosa Cells

COMPARISON OF THE STIMULATORY EFFECTS OF OVINE, PORCINE AND HUMAN FOLLICLE-STIMULATING HORMONE AND OF OVINE AND HUMAN LUTEINIZING HORMONE ON THE ACCUMULATION OF CYCLIC AMP BY PORCINE GRANULOSA CELLS

1979 ◽  
Vol 80 (1) ◽  
pp. 9-20 ◽  
Author(s):  
ADA M. LINDSEY ◽  
CORNELIA P. CHANNING
Keyword(s):  
Cyclic Amp ◽  
Granulosa Cells ◽  
Incubation Medium ◽  
Follicle Stimulating Hormone ◽  
Similar Response ◽  
Intracellular Accumulation ◽  
Porcine Granulosa Cells ◽  
Tenfold Increase ◽  
Incubation Periods

The effects of ovine, porcine and human FSH, and ovine and human LH on the accumulation of cyclic AMP by porcine granulosa cells obtained from follicles at various stages of maturation were investigated. During incubation periods of 15 min, 10 μg ovine FSH pretreated with antiserum to LH or 10 μg human FSH resulted in an 11- to 18-fold, five-to ninefold, and less than a twofold increase in intracellular accumulation of cyclic AMP by granulosa cells from small (1–2 mm), medium (3–5 mm) and large (6–12 mm) follicles respectively. Similar patterns of response occurred with addition of porcine FSH. After incubation for 30 and 60 min with ovine, porcine or human FSH, significant accumulation of cyclic AMP in the incubation medium occurred with cells obtained from small and medium-sized follicles. After 60 min of incubation with FSH the accumulation of cyclic AMP in the incubation medium exceeded the intracellular cyclic AMP levels in granulosa cells from small and medium-sized follicles. During incubation periods of 15 min, 1·0 μg ovine LH resulted in less than a twofold, a fourfold and greater than a tenfold increase in intracellular accumulation of cyclic AMP by granulosa cells from small, medium and large follicles respectively. Addition of human LH brought about a similar response. Incubation periods of 30 and 60 min with 1·0 μg ovine or human LH resulted in significant accumulation of cyclic AMP in the incubation medium by granulosa cells from large follicles; cyclic AMP content in the incubation medium was greater after 60 min compared with 30 min of incubation. It was concluded that ovine FSH pretreated with an antiserum to LH had similar effects on cyclic AMP levels as did purified human and porcine FSH, and that the stimulatory effects of the less pure ovine FSH were probably not due to an impurity in the FSH preparation. Porcine granulosa cells obtained from small follicles should be suitable as an in-vitro FSH bioassay while granulosa cells obtained from large follicles should be suitable as an in-vitro LH bioassay.

scholarly journals Does 2-hydroxyflutamide Inhibit Apoptosis in Porcine Granulosa Cells? ^|^mdash; An In Vitro Study

2012 ◽  
Vol 58 (4) ◽  
pp. 438-444 ◽  
Author(s):  
Malgorzata DUDA ◽  
Malgorzata DURLEJ ◽  
Malgorzata KNET ◽  
Katarzyna KNAPCZYK-STWORA ◽  
Zbigniew TABAROWSKI ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
In Vitro Study ◽  
Vitro Study ◽  
Porcine Granulosa Cells

Secretion of interferon-tau by bovine embryos in long-term culture: comparison of in vivo derived, in vitro produced, nuclear transfer and demi-embryos

1999 ◽  
Vol 55 (3-4) ◽  
pp. 151-162 ◽  
Author(s):  
M Stojkovic ◽  
M Büttner ◽  
V Zakhartchenko ◽  
J Riedl ◽  
H.-D Reichenbach ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Bovine Embryos ◽  
Interferon Tau ◽  
Long Term Culture

Prolactin, LH, and estradiol-17β in utilization of lipoprotein substrate by porcine granulosa cells in vitro

1988 ◽  
Vol 66 (5) ◽  
pp. 561-566 ◽  
Author(s):  
K. Rajkumar ◽  
P. Klingshorn ◽  
P. J. Chedrese ◽  
B. D. Murphy
Keyword(s):  
Granulosa Cell ◽  
Cell Cultures ◽  
Granulosa Cells ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Low Density ◽  
Porcine Granulosa Cells ◽  
Progesterone Synthesis ◽  
Serum Free Conditions

Porcine granulosa cells cultured under serum free conditions responded by increased progesterone secretion to the addition of the leuteotropic hormones, LH, prolactin, and estradiol. Provision of extracellular substrate for steroidogenesis in the form of porcine high density lipoprotein or low density lipoprotein enhanced progesterone accumulation by granulosa cell cultures. Estradiol, LH, and prolactin all greatly increased progesterone accumulation in the presence of either high or low density lipoproteins. Increases in progesterone accumulation following addition of prolactin or LH in combination with estradiol suggested the presence of a synergistic interaction among leuteotropins. Pre-exposure of granulosa cell cultures to estradiol increased the subsequent stimulatory effect of prolactin on lipoprotein utilization. It is concluded that all three leuteotropins function to enhance and may interact in the utilization of extracellular lipoprotein substrate for progesterone synthesis.

scholarly journals Expression of CA-125 by progestational bovine endometrium: prospective regulation and function

Reproduction ◽  
2003 ◽  
pp. 615-620 ◽  
Author(s):  
AC McDonnel ◽  
EA Van Kirk ◽  
KJ Austin ◽  
TR Hansen ◽  
EL Belden ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Ovarian Cancer Cell Line ◽  
Amino Acid Content ◽  
Ca 125 ◽  
Embryonic Cells ◽  
Cancer Antigen 125 ◽  
Interferon Tau ◽  
Endometrial Cells ◽  
And Function

Cancer antigen 125 (CA-125) is expressed by malignant human ovarian surface epithelial cells and derivatives of the Mullerian duct system. This study explored the expression, regulation, and function of CA-125 in the bovine uterus. CA-125 was localized by immunohistochemistry to the apical surfaces of epithelial cells lining the endometrium and proximal glands of the late luteal phase and early pregnancy; antigen was not detected during oestrus or the postpartum period. Production of CA-125 by bovine endometrial cells in vitro was upregulated by progesterone and interferon-tau. Immunopurified CA-125 from uterine flushes of dioestrous or pregnant cows was similar in biochemical composition (as determined by gel electrophoresis and amino acid content) to the human antigen isolated from incubation medium conditioned by the ovarian cancer cell line OVCAR-3. Bovine CA-125 inhibited complement-induced lysis of antibody-sensitized sheep erythrocytes. It is suggested that endometrial CA-125 exerts a progestational role in part by protecting maternal and embryonic cells from immune targeting and lysis.

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