scholarly journals 332IN VIVO DEVELOPMENTAL CAPACITY OF IN VITRO-PRODUCED EMBRYOS DERIVED FROM SEX-SORTED AND RE-CRYOPRESERVED FROZEN-THAWED RAM SPERM

2004 ◽  
Vol 16 (2) ◽  
pp. 286 ◽  
Author(s):  
J.K. O'Brien ◽  
F.K. Hollinshead ◽  
G. Evans ◽  
W.M.C. Maxwell
Keyword(s):  
Potential Application ◽  
High Speed ◽  
Fisher Exact Test ◽  
Embryo Survival ◽  
Food Dye ◽  
Frozen Semen ◽  
Developmental Capacity ◽  
Ram Sperm

The ability to sort and re-freeze frozen-thawed sperm would significantly increase the potential application of sperm sexing technology to species management. Frozen-thawed, sorted, re-frozen and then thawed ram sperm appear fully functional in vitro with blastocyst production greater than that for frozen-thawed, non-sorted sperm (Hollinshead FK et al. 2003 Theriogenology 59, 209 abst). The aim of this study was to evaluate the in vivo capacity of in vitro-produced embryos derived from frozen-thawed sperm after sorting and a second cryopreservation/thawing step. Frozen semen from 2 rams (n=3 ejaculates per ram) was used throughout. Post-thaw sperm treatments comprised (i) non-sorted (Control); (ii) sorted (Froz-Sort) and (iii) sorted, then re-frozen (Froz-Sort-Froz). X and Y sperm were separated using a high-speed sorter (SX MoFlo®, DakoCytomation, Fort Collins, CO, USA) after incubation with Hoechst 33342 and food dye to eliminate nonviable sperm. Reanalysis revealed high levels (mean±SEM) of purity for X- and Y-enriched samples for all treatments (89±1.2%). At Day 6 post-insemination, 2 embryos (blastocyst stage or greater) were transferred per recipient. Data were analyzed by chi-square and Fisher Exact Test. In vivo embryo survival was similar across sperm treatments (28/64, 43.8% overall) and 20 of 23 (87.0%) sexed lambs were of the predicted sex (Table 1). These results demonstrate high in vivo developmental capacity of in vitro-produced sexed embryos derived from frozen-thawed ram sperm after sorting and a second cryopreservation/thawing step, and increase the potential application of sperm sexing technology. Research support by XY, Inc., Australian Research Council and Zoological Parks Board of NSW. Table 1 In vivo survival of transferred in vitro produced embryos derived from frozen-thawed non-sorted (Control), frozen-thawed and sorted (Froz-Sort) and frozen-thawed, sorted, then frozen-thawed (Froz-Sort-Froz) ram sperm.

scholarly journals Birth of lambs of a pre-determined sex after in vitro production of embryos using frozen–thawed sex-sorted and re-frozen–thawed ram spermatozoa

Reproduction ◽  
2004 ◽  
Vol 127 (5) ◽  
pp. 557-568 ◽  
Author(s):  
F K Hollinshead ◽  
G Evans ◽  
K M Evans ◽  
S L Catt ◽  
W M C Maxwell ◽  
...  
Keyword(s):  
High Speed ◽  
Gradient Centrifugation ◽  
In Vitro Production ◽  
Blastocyst Development ◽  
Embryo Survival ◽  
Treatment Groups ◽  
Sperm Migration ◽  
And Control

The characteristics and functional capacity of ram spermatozoa frozen–thawed prior to and after flow cytometric sorting was assessed after incubation (37 °C; 6 h),in vitrofertilisation (IVF), and transfer of fresh and vitrifiedin vitroproduced embryos. Frozen-thawed spermatozoa from two rams were allocated to four treatment groups: (i) non-sorted (Control); (ii) sorted (FS); (iii) sorted then re-frozen (FSF) and (iv) re-frozen control (FCF). Frozen-thawed samples were separated into X- and Y-chromosome bearing spermatozoa using a high-speed sperm sorter after density gradient centrifugation (X: 88 ± 1.5% and Y: 87 ± 1.1% purity). After 6 h incubation (37 °C), the percentage of motile spermatozoa was higher (P< 0.001) for FS (84 ± 2.0%) compared with all other treatments (Control: 36 ± 3.3%, FSF: 28 ± 3.1%, FCF: 20 ± 2.0%). In a sperm migration test greater numbers of FS spermatozoa penetrated 5 mm into the artificial cervical mucus compared with spermatozoa from all other treatments (152 ± 39.4 vs 31 ± 9.2 spermatozoa respectively;P< 0.05). Fertilisation and cleavage rates were higher (P< 0.05) forin vitromatured oocytes inseminated with Control compared with FSF spermatozoa. However, the Day 7 blastocyst development rate was higher for oocytes inseminated with FSF (62.2%) than FS and Control spermatozoa (52.7 and 50.0%;P< 0.05). The number of ewes pregnant (Day 60), lambing and thein vivoembryo survival rate was greater (P< 0.01) after the transfer of fresh embryos rather than vitrified embryos derived from X- and Y-spermatozoa (67.6, 64.7 and 41.2% vs 29.6, 25.9 and 14.8% respectively). Twenty-six of the 30 (86.7%) lambs derived from sex-sorted spermatozoa were of the correct sex. These results demonstrate that frozen–thawed ram spermatozoa can be sex-sorted for immediate or future use after re-cryopreservation and, in conjunction with IVF and embryo transfer, can be used to efficiently produce offspring of pre-determined sex.

229 OOCYTE RECOVERY, IN VITRO FERTILIZATION AND EMBRYO TRANSFER IN THE SERVAL (LEPTAILURUS SERVAL)

2006 ◽  
Vol 18 (2) ◽  
pp. 223 ◽  
Author(s):  
C. E. Pope ◽  
M. C. Gómez ◽  
A. Cole ◽  
C. Dumas ◽  
B. L. Dresser
Keyword(s):  
Embryo Transfer ◽  
Oocyte Retrieval ◽  
Sub Saharan Africa ◽  
Medium Size ◽  
Embryo Survival ◽  
Frozen Semen ◽  
Old Donor ◽  
Fresh Embryos

Servals are medium size (9 to 18 kg) spotted cats found in sub-Saharan Africa that are protected by CITES under Appendix II regulations. There are at least six sub-species, one of which is listed as Endangered by the U.S. Endangered Species Act. In vitro-derived embryos have been produced in at least one-half of the 36 species of nondomestic cats, and kittens have been born after embryo transfer in six species. In the present study we evaluated (1) ovarian response of servals to repeated exogenous gonadotropin stimulation, and (2) in vitro and in vivo developmental ability of in vitro-derived embryos. One two-year-old and one five-year-old female were treated six and three times, respectively, over a 3.5-year period, with a total of 20 or 25 IU of porcine FSH (i.m.; Sioux Biochem, Sioux City, IA, USA) administered daily over four days during interestrus. On Day 5, 15 IU of porcine LH (i.m.; Sioux Biochem) was given, and laparoscopic oocyte retrieval was performed 24 h later. A total of 234 preovulatory oocytes were recovered: 182 (mean = 30.3) from the two-year-old and 52 (mean = 17.3) from the five-year-old female. A total of 91 and 91 oocytes were recovered at retrievals 1 through 3 and 4 through 6, respectively, from the two-year-old donor. Eighty oocytes from the two-year-old donor were inseminated with cooled (24 h, 4°C) semen. Frozen semen from the same male was used to inseminate 102 oocytes from the two-year-old female and 52 oocytes from the five-year-old female. Overall, 136 embryos (58% cleavage frequency) were produced: 119 (65% cleavage frequency) from the two-year-old and 17 (33% cleavage frequency) from the five-year-old female. Cleavage frequency of oocytes from the two-year-old female inseminated with cooled or frozen semen was similar, 68% (54/80) and 64% (65/102), respectively. Embryos were cultured for 5 or 6 days before controlled rate cryopreservation or uterine transfer (Gómez et al. 2003 Theriogenology 60, 239–251). On Day 5, 66 early to mid-stage morulae were cryopreserved at a slow controlled rate. Sixty Day 5 and 18 Day 6 embryos were auto-transferred to a recipient (8 to 26/transfer) in a total of six surgical procedures, of which five were with fresh embryos (n = 70) and one was with cryopreserved embryos (n = 8). The sixth embryo transfer procedure (26 fresh embryos) resulted in the unassisted birth of a live male kitten on Day 77 of gestation. We have shown that in vitro-derived embryos can be generated in the serval and that oocyte retrieval rates and cleavage frequencies are comparable to those reported for other species of mid-sized nondomestic cats. The nominal incidence of pregnancy and frequency of embryo survival may be improved by transferring early cleavage staged embryos into the oviduct, as demonstrated in the African wildcat (Felis silvestris lybica; Gómez et al. 2004 Cloning and Stem Cells 6, 247–258). This work was partially funded by the Dan Heard Conservation Challenge Grant.

scholarly journals 173TRANSFER OF VITRIFIED BLASTOCYSTS FROM ONE OR TWO SUPEROVULATED LARGE WHITE HYPERPROLIFIC DONORS TO MEISHAN RECIPIENTS: REPRODUCTIVE PARAMETERS AT DAY 30 OF PREGNANCY

2004 ◽  
Vol 16 (2) ◽  
pp. 208
Author(s):  
C. Cuello ◽  
F. Berthelot ◽  
F. Martinat-Botté ◽  
P. Guillouet ◽  
V. Furstoss ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Fisher Exact Test ◽  
Corpora Lutea ◽  
Large White ◽  
Embryo Survival ◽  
Exact Test ◽  
Group 2 ◽  
Fresh Semen ◽  
Group 1

The present study was designed to determine the effect of pooling embryos from two donors on the reproductive success of transfer of vitrified/warmed porcine blastocysts. Superovulated Large White hyperprolific gilts (n=24) were used as embryo donors. Gilts were artificially inseminated 12 and 24h after initial detection of estrus using fresh semen, and slaughtered on Days 5.5 to 6 of the estrous cycle (Day 0=Onset of estrus). Embryos were recovered by flushing the uterine horns, and unhatched blastocysts were selected. Vitrification and warming were performed as reported previously (Berthelot et al., 2000 Cryobiology 41, 116–124). Embryo transfers were conducted in asynchronous (−24h) Meishan gilts (n=20). Twenty vitrified/warmed blastocysts were surgically transferred into one uterine horn. Ten recipients received embryos from one donor (group 1) and the other ten transfers were performed with mixed embryos from two donors (group 2). Pregnancy was assessed ultrasonographically at Day 25 after estrus and recipients were slaughtered five days later. The pregnancy rate from the different groups was compared using Fisher exact test. The GLM procedure of SAS was used to determine the effect of the origin of embryos (one or two donors) on the number of developed fetuses and viable fetuses at Day 30 of pregnancy. The ovulation rate was 32.5±11.8 (mean±SD). The total number of embryos collected was 634, of which 57 (9.0%), 36 (5.7%), 513 (80.9%) and 28 (4.4%), were unfertilized oocytes and degenerated embryos, morulae, unhatched blastocysts and hatched blastocysts, respectively. The ratio of collected embryos to the number of corpora lutea was 81.3%. The pregnancy rate for group 1 (70%) was not different (P&gt;0.05) than that for group 2 (90%). No significant differences were detected between group 1 and group 2 for in vivo embryo development (number fetuses/transferred embryos in pregnant recipients; 33.3% v. 40%) or in vivo embryo survival (number viable fetuses/transferred embryos in pregnant recipients; 27.9% v. 33.9%). However, the in vivo efficiency (number viable fetuses/total transferred embryos) was higher (P&lt;0.05) when transfers were performed with embryos from two donors (19.5% v. 30.5%). These results indicate that pooling embryos from two donors increases the in vivo efficiency after transfer of vitrified/warmed porcine blastocysts. This study was supported by grant from SENECA (FPI/99, Spain).

scholarly journals 183PREGNANCY RATE FOLLOWING TRANSFER OF IN VITRO- AND IN VIVO-PRODUCED BOVINE EMBRYOS TO LH-TREATED RECIPIENTS

2004 ◽  
Vol 16 (2) ◽  
pp. 213 ◽  
Author(s):  
J. Small ◽  
M. Colazo ◽  
D. Ambrose ◽  
R. Mapletoft ◽  
J. Reeb ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Animal Health ◽  
Beef Cows ◽  
Water Bath ◽  
Bovine Embryos ◽  
Chi Square ◽  
Embryo Survival ◽  
Frozen Embryos

The objective was to evaluate the effect of pLH treatment on pregnancy rates in recipients receiving in vivo- or in vitro-produced bovine embryos. Heifers (n=37) and lactating (n=28) and non-lactating (n=150) beef cows were treated at random stages of the cycle with 100μg GnRH i.m. (Cystorelin, Merial Canada Inc., Victoriaville, Quebec, Canada) on Day −9, 500μg cloprostenol i.m. (PGF; Estrumate, Schering Plough Animal Health, Pointe-Claire, Quebec, Canada) on Day —2 and GnRH on Day 0 (66h post-PGF; without estrus detection). Cattle were placed at random, by class, into three groups: no further treatment (Control; n=71), or 12.5mg pLH (Lutropin-V, Bioniche Animal Health, Belleville, Ontario, Canada) on Day 5 (n=72) or on Day 7 (n=72) after the second GnRH. On Day 7, cattle with a CL &gt;10mm in diameter (determined ultrasonically) received in vivo-produced, fresh (Simmental) or frozen (Holstein), or in vitro-produced frozen (Holstein) embryos (embryo type balanced among groups). Embryos were cryopreserved in 10% ethylene glycol; in vivo-produced frozen embryos were thawed 5 to 10s in air, 15s in a water-bath at 30°C and then “direct-transferred” nonsurgically. In vitro-produced frozen embryos (donated by IND Lifetech Inc., Delta, British Columbia, Canada) were thawed in a water-bath at 27°C for 10s and placed in ViGro Holding Plus medium (AB Technology, Pullman, WA, USA) at room temperature, evaluated and then transferred nonsurgically. Pregnancy was determined by ultrasonography on Day 35. Data were analyzed with CATMOD, chi-square and GLM procedures (SAS Institute, Cary, NC, USA.). Twenty cattle (9.3%) did not receive embryos; five heifers had cervical problems, and five heifers and 10 cows did not have a CL &gt;10mm. Overall, 7.1% of the recipients had two CL on the day of embryo transfer. There was no effect (P&gt;0.05) of treatment, embryo type (or interaction) or class of recipient on pregnancy rate (overall, 44.1%, 86/195; Table 1). Similarly, mean (±SD) CL diameter and luteal area did not differ (P&gt;0.05) among groups or between pregnant and open recipients (overall, 22.0±3.4mm and 352.0±108.7mm, respectively). However, recipients with a CL diameter ≥18mm tended (P&lt;0.1) to have a higher pregnancy rate (45.8 vs 25.0%). In a subset of 40 recipients examined ultrasonically on Day 12, 50% of those treated on Day 5 and 70% of those treated with pLH on Day 7 had two CL. In summary, overall pregnancy rate in GnRH-synchronized recipients receiving in vitro- or in vivo-produced embryos by nonsurgical transfer was 44.1%. Embryo survival to Day 35 was not affected by type of embryo or treatment with pLH 5 or 7 days after ovulation. Table 1 Pregnancy rate in recipients on Day 35 based on pLH treatment and embryo-type

scholarly journals Coherent Raman Scattering Microscopy in Oncology Pharmacokinetic Research

2021 ◽  
Vol 12 ◽  
Author(s):  
Junjie Zeng ◽  
Wenying Zhao ◽  
Shuhua Yue
Keyword(s):  
Raman Scattering ◽  
High Speed ◽  
Cancer Drug ◽  
Cancer Drugs ◽  
High Speed Imaging ◽  
Coherent Raman Scattering ◽  
Anti Cancer ◽  
Coherent Raman

The high attrition rates of anti-cancer drugs during clinical development remains a bottleneck problem in pharmaceutical industry. This is partially due to the lack of quantitative, selective, and rapid readouts of anti-cancer drug activity in situ with high resolution. Although fluorescence microscopy has been commonly used in oncology pharmacological research, fluorescent labels are often too large in size for small drug molecules, and thus may disturb the function or metabolism of these molecules. Such challenge can be overcome by coherent Raman scattering microscopy, which is capable of chemically selective, highly sensitive, high spatial resolution, and high-speed imaging, without the need of any labeling. Coherent Raman scattering microscopy has tremendously improved the understanding of pharmaceutical materials in the solid state, pharmacokinetics of anti-cancer drugs and nanocarriers in vitro and in vivo. This review focuses on the latest applications of coherent Raman scattering microscopy as a new emerging platform to facilitate oncology pharmacokinetic research.

Effect of dietary energy concentrations during the peri-ovulatory period on the in vivo and in vitro development of fertilized sheep ova

1995 ◽  
Vol 1995 ◽  
pp. 74-74
Author(s):  
N.M. Al-Khozam ◽  
J.J. Robinson ◽  
T.G. McEvoy ◽  
R.P. Aitken ◽  
P.A. Findlay ◽  
...  
Keyword(s):  
Subsequent Development ◽  
Published Data ◽  
Arid Environments ◽  
Dietary Energy ◽  
Embryo Survival ◽  
Short Supply ◽  
Ovulatory Period ◽  
Vitro Development

Results from a series of recent experiments involving superovulated ewes demonstrate the important influence of nutritionally-induced alterations in preovulatory progesterone concentrations on the subsequent in vivo and in vitro development of their fertilized ova (McEvoy et al, 1993 and 1995; Creed et al, 1994). In essence, these show that high-plane feeding can suppress preovulatory progesterone concentrations to such an extent that the subsequent development of the ova is impaired both in vivo and during in vitro culture. An important practical question however remains unanswered in that no attempt has been made to study the effects of dietary energy concentrations, as opposed to plane-of-nutrition, on progesterone concentrations and ovum development. As a result, recommendations regarding which energy sources should be used as supplements to pasture around mating time are a matter of conjecture. Furthermore, in arid environments, roughage feeds are often in short supply and therefore command a much higher price per unit of energy than concentrate diets. Under these conditions it is not unusual to feed all-concentrate diets at mating, yet there are no published data for their effects on ovum development and embryo survival.

Endocrine and physiological events from ovulation to establishment of pregnancy in cattle

2001 ◽  
Vol 26 (1) ◽  
pp. 81-91 ◽  
Author(s):  
W.W. Thatcher ◽  
M. Binelli ◽  
D. Arnold ◽  
R. Mattos ◽  
L. Badinga ◽  
...  
Keyword(s):  
Dairy Cows ◽  
Post Partum ◽  
Secretory Response ◽  
Embryo Survival ◽  
Endometrial Cells ◽  
Stat Proteins ◽  
In Vivo Experiments ◽  
Feeding Diets

AbstractA series of in vitro and in vivo experiments were conducted to characterise the dialogue between embryo and maternal units relative to the mechanisms controlling embryo survival in dairy cattle. Endometrial explants from pregnant cows had an attenuated PGF2α secretory response following treatment with melittin (stimulator of PLA2) and phorbol 12, 13 dibutyrate (PDBu). Thus previous exposure to the conceptus appears to regulate the endometrial synthetic pathway at a point coincident with or distal to PLA2 as well as inhibit PKC or PKC mediated events. Endometrial explants collected from cows receiving intrauterine infusions of rblFN-τ had a reduced secretory response following stimulation with PDBu indicating attenuation in PKC activity. Based upon tyrosine-phosphorylation of STAT-proteins and their translocation to the nucleus after treatment with rbIFN-τ, the JAK-STAT pathway is functional in immortalised bovine endometrial cells (BEND cells). Bend cells, exposed to rblFN-τ, reduced PDBu induction of PGF2α secretion and also decreased protein expression of Cox-2 and PLA. RblFN-τ clearly reduced PKC mediated events leading to an antiluteolytic response in endometrial cells. Feeding diets containing 2.6, 5.2 and 7.8% Menhaden fish meal to lactating dairy cows reduced uterine secretion of PGF2α following sequential injections of oestradiol and oxytocin. Thus antiluteolytic effects in early pregnancy may be amplified by feeding by-pass fats. Pregnancy rate to a timed insemination at first service post-partum is increased in association with injection of bST(500 mg; sc) given at insemination. Furthermore injection of bST at time of insemination in superovulated donor cows increased the number of blastocysts and reduced number of unfertilised embryos. Prospects of integrating novel strategies to improve embryo development and survival into reproductive management systems appear to be attainable in high producing dairy cows.

scholarly journals Birth of a Live Cria After Transfer of a Vitrified-Warmed Alpaca (Vicugna pacos) Preimplantation Embryo

2020 ◽  
Vol 7 ◽  
Author(s):  
Jennifer C. Lutz ◽  
Susan L. Johnson ◽  
Kimberly J. Duprey ◽  
Paul J. Taylor ◽  
Henry William Vivanco-Mackie ◽  
...  
Keyword(s):  
Ethylene Glycol ◽  
Preimplantation Embryos ◽  
Embryo Survival ◽  
Important Species ◽  
Vicugna Pacos ◽  
Improvement Programs ◽  
The One ◽  
Humidified Air

The alpaca (Vicugna pacos) is an important species for the production of fiber and food. Genetic improvement programs for alpacas have been hindered, however, by the lack of field-practical techniques for artificial insemination and embryo transfer. In particular, successful techniques for the cryopreservation of alpaca preimplantation embryos have not been reported previously. The objective of this study was to develop a field-practical and efficacious technique for cryopreservation of alpaca preimplantation embryos using a modification of a vitrification protocol originally devised for horses and adapted for dromedary camels. Four naturally cycling non-superovulated Huacaya females serving as embryo donors were mated to males of proven fertility. Donors received 30 μg of gonadorelin at the time of breeding, and embryos were non-surgically recovered 7 days after mating. Recovered embryos (n = 4) were placed individually through a series of three vitrification solutions at 20°C (VS1: 1.4 M glycerol; VS2: 1.4 M glycerol + 3.6 M ethylene glycol; VS3: 3.4 M glycerol + 4.6 M ethylene glycol) before loading into an open-pulled straw (OPS) and plunging directly into liquid nitrogen for storage. At warming, each individual embryo was sequentially placed through warming solutions (WS1: 0.5 M galactose at 37°C; WS2: 0.25 M galactose at 20°C), and warmed embryos were incubated at 37°C in 5% CO2 in humidified air for 20–22 h in 1 ml Syngro® holding medium supplemented with 10% (v/v) alpaca serum to perform an initial in vitro assessment of post-warming viability. Embryos whose diameter increased during culture (n = 2) were transferred individually into synchronous recipients, whereas embryos that did not grow (n = 2) were transferred together into a single recipient to perform an in vivo assessment of post-warming viability. Initial pregnancy detection was performed ultrasonographically 29 days post-transfer when fetal heartbeat could be detected, and one of three recipients was pregnant (25% embryo survival rate). On November 13, 2019, the one pregnant recipient delivered what is believed to be the world's first cria produced from a vitrified-warmed alpaca embryo.

230 GENERATION OF PUTATIVE RABBIT EMBRYONIC STEM CELLS

2007 ◽  
Vol 19 (1) ◽  
pp. 231
Author(s):  
S. Wang ◽  
X. Tang ◽  
Y. Niu ◽  
H. Chen ◽  
T. Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
High Speed ◽  
Es Cells ◽  
Research Work ◽  
Embryonic Stem ◽  
Morphological Characteristics ◽  
Mouse Escs

The rabbit, as a laboratory animal model, has several advantages in the study of human physiological disorders. In this study, stable putative pluripotent rabbit embryonic stem cells (rESCs) were derived from in vivo-fertilized and in vitro-cultured blastocysts. The rabbit ICMs were obtained by 0.05% trypsin–0.008% EDTA treatment and mechanical separation; the ES-like cell colonies seen several days later. ICM-derived outgrowths which were treated with 5 mg/mL-1 dispase, followed by 0.05% trypsin–0.008% EDTA, were mechanically disaggregated into small clumps and reseeded on MEFs. The putative ES cell lines maintained expression of pluripotent cells markers and normal XY karyotype for long periods of culture (&gt;1 month). The putative rESCs expressed alkaline phosphatase, transcription factor Oct-4, stage-specific embryonic antigens (SSEA-1, SSEA-3, and SSEA-4), and tumor-related antigens (TRA-1-60 and TRA-1-81). The morphological characteristics of the putative ESCs are closer to those of human ESCs; their high speed of proliferation, however, is closer to that of mouse ESCs. Putative rabbit ESCs were induced to differentiate into many cell types including trophoblast cells, similar to primate ESCs, in vitro, and formed teratomas with derivatives of the 3 major germ layers in vivo when injected into SCID mice. Using RT-PCR measurement, but with some differences in ligands and inhibitors, and comparing with human and mouse ESCs, the putative rabbit ESCs expressed similar genes related to pluripotency (Oct-4, Nanog, SOX2, and UTF-1) and similar genes of FGF, WNT, and TGF signaling pathways related to the proliferation and self-renewal. Our further research work showed that TGF beta and FGF pathways cooperate to maintain pluripotency of rabbit ESCs similar to those of human ES cells.

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