138 COMPARISON OF KINETICS AND PATTERNS OF THE FIRST CLEAVAGE OF IN VIVO AND IN VITRO-MATURED HOLSTEIN OOCYTES AFTER IN VITRO FERTILIZATION WITH X-SORTED SPERM

2014 ◽  
Vol 26 (1) ◽  
pp. 182
Author(s):  
S. Matoba ◽  
S. Sugimura ◽  
H. Matsuda ◽  
Y. Aikawa ◽  
M. Ohtake ◽  
...  
Keyword(s):  
Calf Serum ◽  
Time Lapse ◽  
Developmental Competence ◽  
High Rate ◽  
Holstein Cows ◽  
Group A ◽  
Group B ◽  
Group D

Previously, it was reported that a high rate of good quality blastocysts were produced by IVF of in vivo-matured oocytes, obtained by ovum pick up (OPU) after superstimulation in Holstein cows, using X-sorted sperm (Matoba et al. 2012 Reprod. Domest. Anim. 47, 515). In this system, an early first cleavage within 28 h after IVF was found to be a potent marker for the selection of embryos with high developmental competence (Matoba et al. 2013 Reprod. Fertil. Dev. 25, 266). However, we have limited knowledge on the timing and normality of embryonic cleavages in in vitro-matured oocytes after IVF. The purpose of the present study was to compare the kinetics and patterns of the first cleavage of in vivo- and in vitro-matured bovine oocytes after IVF with X-sorted sperm. In vivo-matured oocytes (Group A) were collected by OPU from non-lactating Holstein cows just before ovulation after superstimulation. Immature oocytes were either collected by OPU without hormonal treatment or by aspiration of ovaries at the local abattoir and matured in vitro (Group B or C). All the oocytes were inseminated with 5 × 106 sperm mL–1 of X-sorted sperm, except half of oocytes in Group C inseminated by non-sorted sperm (Group D) and cultured in CR1aa supplemented with 5% calf serum and 0.25 mg mL–1 of linoleic acid albumin at 38.5°C in 5% CO2, 5% O2, and 90% N2 for 216 h. Embryo kinetics were observed individually using a microwell culture dish and time-lapse cinematography (Sugimura et al. 2010 Biol. Reprod. 83, 970–978). Photographs of each embryo were taken in every 15 min during the IVC period and analysed by time-lapse cinematography software. Cleavage pattern was categorized as normal (2 even blastomeres without fragment or protrusion) or abnormal (2 uneven blastomeres, with fragment or protrusion and those dividing into 3–5 blastomeres) at the first cleavage. Data were analysed by ANOVA, chi-squared, or discriminant function. A total of 268 cleaved embryos were used. The blastocyst rate in Group A was higher than in Groups B and C (61.3 v. 40.0 and 25.0%, respectively; P < 0.05). The timing of first cleavage was longer in Group A compared with Groups C and D (28.3 ± 3.8 v. 27.6 ± 3.8 and 26.7 ± 1.9 h, respectively) and in Group B (28.1 ± 4.0 h) compared with in Group D (P < 0.05). Higher rates of normal cleavage were observed in Groups A, B, and D than in Group C (53.5, 44.4, and 54.8 v. 16.7%, respectively; P < 0.01). The frequency of blastocysts derived from the early (28.3 h) and normal pattern cleaving oocytes were greater in Groups A and B than in Group C (29.0 and 20.0 v. 8.3%, respectively; P < 0.05) and similar in Group D (22.6%). Our results reveal that IVF embryos produced from in vivo-matured oocytes with sex-sorted sperm had superior normality than those produced from in vitro-matured oocytes and similar normality to embryos inseminated with non-sorted sperm. Supported by the Research and Development projects for application in promoting new policy of agriculture, forestry and fisheries (22016) and by JSPS and HAS under the Japan-Hungary Research Cooperative Program.

236 KINETICS AND PATTERN OF THE FIRST CLEAVAGE OF IN VITRO-FERTILIZED EMBRYOS BY IN VIVO-MATURED OOCYTES AND X-SORTED SPERMATOZOA IN DAIRY CATTLE

2013 ◽  
Vol 25 (1) ◽  
pp. 266
Author(s):  
S. Matoba ◽  
S. Sugimura ◽  
H. Matsuda ◽  
Y. Aikawa ◽  
M. Ohtake ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Calf Serum ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Holstein Cows ◽  
Cleavage Pattern ◽  
Cell Cleavage ◽  
Significant Difference

Recently, we reported that high rates of good-quality blastocysts can be produced by IVF of in vivo-matured oocytes, obtained by ovum pick-up (OPU) after superstimulation in Holstein cows, with X-sorted sperm [Matoba et al. 2012 Reprod. Domest. Anim. 47(Suppl. 4), 515]. However, we have limited knowledge concerning the normality of embryonic cleavages in such embryos. The present study examined their kinetics and pattern of the first cell cycle. In vivo-matured oocytes were collected by OPU from non-lactating Holstein cows just before ovulation after superstimulation and ovulation induction by gonadotropin-releasing hormone. The oocytes were inseminated with 5 × 106 sperm mL–1 of X-sorted sperm and cultured in CR1aa supplemented with 5% newborn calf serum and 0.25 mg mL–1 of linoleic acid albumin at 38.5°C in 5% CO2, 5% O2, and 90% N2 for 216 h. Embryo kinetics were observed individually using a microwell culture dish and time-lapse cinematography (CCM-1.4MZS, Astec, Fukuoka, Japan) (Sugimura et al. 2010 Biol. Reprod. 83, 970–978). Photographs of each embryo were taken every 15 min during the in vitro culture period, and images were analysed by CCM-1.4 software (Astec). The cleavage pattern was categorised into normal cleavage (2 even blastomeres without fragment or protrusion) or abnormal cleavage (those with 2 uneven blastomeres, with fragments or protrusions and those dividing into 3 to 5 blastomeres at the first cleavage). Data were analysed by ANOVA, chi-square, and discriminant function. A total of 117 embryos were examined; of this number, 63.2% developed to the blastocyst stage and the rest were degenerated. A high rate of normal cleavage and a low rate of abnormal cleavage, including those with 2 uneven blastomeres and those with fragments or protrusions in the first cleavage pattern, were recorded in embryos that could develop to blastocysts compared with degenerated ones (P < 0.01 or P < 0.05, respectively; Table 1). No significant difference was found in those dividing into 3 to 5 blastomeres between the blastocysts and degenerated embryos (Table 1). Embryos developing to the blastocyst stage had a shorter duration of the first cell cycle [27.2 ± 2.3 h post-insemination (hpi)] compared with those undergoing degeneration (30.6 ± 5.7 hpi; P < 0.001). The threshold of duration of the first cell cycle was calculated by (X – 27.2)/2.3 = (30.6 – X)/5.7, resulting in X = 28.2. Blastocysts with a short duration of the first cell cleavage (≤28.2 hpi) showed a higher frequency of the normal cleavage pattern than those with a duration of the first cell cleavage longer than 28.2 hpi (71.7 and 53.6%, respectively; P < 0.05). Our results revealed that those IVF embryos that finished their first cleavage before 28.2 h of IVF and showed a normal cleavage pattern had superior developmental competence. Table 1.The first cleavege pattern reflects the developmental competence: blastocysts versus degenerated embryos This work was supported by the Research and Development Projects for Application in Promoting New Policy of Agriculture, Forestry and Fisheries (22016).

scholarly journals Intrahepatic Arterial Delivery of Sorafenib Eluting Beads: A Pharmacokinetics Study

2020 ◽  
Vol 3 (2) ◽  
pp. 17
Author(s):  
Eerdunbagena Ning ◽  
Zhijun Wang
Keyword(s):  
Liver Tissue ◽  
Slow Release ◽  
Normal Liver ◽  
In Vivo Study ◽  
Pharmacokinetics Study ◽  
Group A ◽  
Group B ◽  
Group D

 Objective: To determine the slow-release effect of Sorafenib carried beads and its impact on the normal liver of dogs. Materials and Methods: (1) To obtain the maximal drug-carrying of beads, different sizes of beads (300-500 μm and 500-700 μm) were tried. Five bottles of different sizes of beads were added into 75% solution of Sorafenid-alcohol with different concentrations: Bottle a,50mg/20ml; Bottle b, 100mg/20ml; Bottle c, 100 mg/40ml; Bottle d, 200mg/40m; Bottle e, 250mg/50ml. (2) In vivo study: 12 dogs were randomly divided into four groups [group A, Sorafenib carried bead (500-700μm); group B, only bead (300-500μm) ; group C, Lipiodol-sorafenib and four dogs in each group. Each group was treated with TAE with emulsion mentioned above. Sorafenib concentration in plasma and liver tissue was determined with HPLC respectively. Result: (1) In vitro research: Sorafenib can be dissolved into 75% alcohol and the best concentration for drug-carrying was 100mg/20ml. (2) In vivo study: ① Compared with group D, the Cmax and AUC in plasma in group A and B has a significant statistics difference(p<0.05). ② Sorafenib concentration in liver tissue could be determined in group A in the 3rd day and even after one week while it could not be determined in group D. Conclusion: Sorafenib can be carried in DC-Bead in a certain condition. Compared with emulsion with Sorafenib and lipiodol, DC-bead has a definite slow-release function and it is superior to lipiodol.

scholarly journals Chromatin organization and timing of polar body I extrusion identify developmentally competent mouse oocytes

2019 ◽  
Vol 63 (3-4-5) ◽  
pp. 245-251 ◽  
Author(s):  
Federica Cavalera ◽  
Mario Zanoni ◽  
Valeria Merico ◽  
Lucia Sacchi ◽  
Riccardo Bellazzi ◽  
...  
Keyword(s):  
Dna Binding ◽  
Polar Body ◽  
Time Lapse ◽  
Reproductive Technologies ◽  
Developmental Competence ◽  
Cell Stage ◽  
Germinal Vesicle Breakdown ◽  
Group A ◽  
Group B

In the mouse, the use of the DNA-binding fluorochrome Hoechst 33342 allows the classification of fully-grown antral oocytes into two categories distinguished by their chromatin conformation: surrounding nucleolus (SN) and not-surrounding nucleolus (NSN) oocytes, the former capable of completing development, the latter unable to proceed beyond the 2-cell stage. In the present study, time-lapse observation of SN and NSN oocyte GV-to-MII transition highlighted differences in the timing of germinal vesicle breakdown (GVBD) and polar body I (PB-I) extrusion. PB-I extrusion, but not GVBD, revealed the presence of three main groups of significantly different oocytes: Group A (456-576 min) comprising mainly SN oocytes (91.4%), group B (584-728 min) entailing an almost equivalent percentage of SN (52.7%) and NSN (47.3%) oocytes, whereas group C (736-896 min) consisting of almost all NSN (94.4%) oocytes. In a further set of time-lapse experiments, GV oocytes were in vitro matured without Hoechst staining and, depending on the timing of PB-I extrusion, sorted into group A, B or C, inseminated with sperm and observed throughout preimplantation. The results show that 26.2 &plusmn; 12.3% of group A, 2.4 &plusmn; 5.0% of group B and none of group C MII oocytes developed to blastocyst. Overall, this study shows that SN oocytes that complete MI earlier are those with a better developmental competence. The possibility to avoid the use of the invasive DNA-binding fluorochrome Hoechst is relevant for future applications in human and domestic animal reproductive technologies.

scholarly journals Photopolymerizable Injectable Cartilage Mimetic Hydrogel for the Treatment of Focal Chondral Lesions: A Proof of Concept Study in a Rabbit Animal Model

2018 ◽  
Vol 47 (1) ◽  
pp. 212-221 ◽  
Author(s):  
Cecilia Pascual-Garrido ◽  
Elizabeth A. Aisenbrey ◽  
Francisco Rodriguez-Fontan ◽  
Karin A. Payne ◽  
Stephanie J. Bryant ◽  
...  
Keyword(s):  
Animal Model ◽  
Chondrogenic Differentiation ◽  
Osteochondral Lesions ◽  
Chondral Defect ◽  
Group A ◽  
Group B ◽  
Safranin O

Background: In this study, we investigate the in vitro and in vivo chondrogenic capacity of a novel photopolymerizable cartilage mimetic hydrogel, enhanced with extracellular matrix analogs, for cartilage regeneration. Purpose: To (1) determine whether mesenchymal stem cells (MSCs) embedded in a novel cartilage mimetic hydrogel support in vitro chondrogenesis, (2) demonstrate that the proposed hydrogel can be delivered in situ in a critical chondral defect in a rabbit model, and (3) determine whether the hydrogel with or without MSCs supports in vivo chondrogenesis in a critical chondral defect. Study Design: Controlled laboratory study. Methods: Rabbit bone marrow–derived MSCs were isolated, expanded, encapsulated in the hydrogel, and cultured in chondrogenic differentiation medium for 9 weeks. Compressive modulus was evaluated at day 1 and at weeks 3, 6, and 9. Chondrogenic differentiation was investigated via quantitative polymerase reaction, safranin-O staining, and immunofluorescence. In vivo, a 3 mm–wide × 2-mm-deep chondral defect was created bilaterally on the knee trochlea of 10 rabbits. Each animal had 1 defect randomly assigned to be treated with hydrogel with or without MSCs, and the contralateral knee was left untreated. Hence, each rabbit served as its own matched control. Three groups were established: group A, hydrogel (n = 5); group B, hydrogel with MSCs (n = 5); and group C, control (n = 10). Repair tissue was evaluated at 6 months after intervention. Results: In vitro, chondrogenesis and the degradable behavior of the hydrogel by MSCs were confirmed. In vivo, the hydrogel could be delivered intraoperatively in a sterile manner. Overall, the hydrogel group had the highest scores on the modified O’Driscoll scoring system (group A, 17.4 ± 4.7; group B, 13 ± 3; group C, 16.7 ± 2.9) ( P = .11) and showed higher safranin-O staining (group A, 49.4% ± 20%; group B, 25.8% ± 16.4%; group C, 36.9% ± 25.2%) ( P = .27), although significance was not detected for either parameter. Conclusion: This study provides the first evidence of the ability to photopolymerize this novel hydrogel in situ and assess its ability to provide chondrogenic cues for cartilage repair in a small animal model. In vitro chondrogenesis was evident when MSCs were encapsulated in the hydrogel. Clinical Relevance: Cartilage mimetic hydrogel may offer a tissue engineering approach for the treatment of osteochondral lesions.

Effects of bovine oviduct epithelial cells, fetal calf serum and bovine serum albumin on gene expression in single bovine embryos produced in the synthetic oviduct fluid culture system

2005 ◽  
Vol 17 (8) ◽  
pp. 751 ◽  
Author(s):  
Mona E. Pedersen ◽  
Øzen Banu Øzdas ◽  
Wenche Farstad ◽  
Aage Tverdal ◽  
Ingrid Olsaker
Keyword(s):  
Gene Expression ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Developmental Competence ◽  
Bovine Embryos ◽  
Glut 1 ◽  
Significant Difference ◽  
Oviduct Fluid

In this study the synthetic oviduct fluid (SOF) system with bovine oviduct epithelial cell (BOEC) co-culture is compared with an SOF system with common protein supplements. One thousand six hundred bovine embryos were cultured in SOF media supplemented with BOEC, fetal calf serum (FCS) and bovine serum albumin (BSA). Eight different culture groups were assigned according to the different supplementation factors. Developmental competence and the expression levels of five genes, namely glucose transporter-1 (Glut-1), heat shock protein 70 (HSP), connexin43 (Cx43), β-actin (ACTB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), analysed as mRNA by using reverse transcription–polymerase chain reaction, were measured on bovine embryos cultured for 9 days. Gene expression of these in vitro-produced embryos was compared with the gene expression of in vivo-produced embryos. There was no significant difference found in embryo developmental competence between the Day 9 embryos in BOEC co-culture, FCS and BSA supplements in SOF media. However, differences in gene expression were observed. With respect to gene expression in in vivo and in vitro embryos, BOEC co-culture affected the same genes as did supplementation with FCS and BSA. HSP was the only gene that differed significantly between in vitro and in vivo embryos. When the different in vitro groups were compared, a significant difference between the BOEC co-culture and the FCS supplementation groups due to Glut-1 expression was observed.

98 SUCCESSFUL PREGNANCIES FOLLOWING TRANSFER OF VITRIFIED PORCINE EMBRYOS DERIVED FROM IN VITRO-MATURED OOCYTES

2006 ◽  
Vol 18 (2) ◽  
pp. 157 ◽  
Author(s):  
K. Hiruma ◽  
H. Ueda ◽  
H. Saito ◽  
C. Tanaka ◽  
N. Maeda ◽  
...  
Keyword(s):  
Calf Serum ◽  
Developmental Competence ◽  
Penicillin G ◽  
Cell Stage ◽  
Epidermal Growth ◽  
Potassium Penicillin ◽  
The One ◽  
Parthenogenetic Embryos

To date only in vivo-produced embryos have successfully produced live piglets after cryopreservation. In this study, we aimed to produce piglets from vitrified embryos derived from in vitro matured (IVM) oocytes. Cumulus-oocyte complexes collected from ovaries obtained at a local slaughterhouse were matured for 44 to 45 h in NCSU23 MEDIUM supplemented with 0.6 mM cysteine, 10 ng/mL epidermal growth factor, 10% (v/v) porcine follicular fluid, 75 �g/mL potassium penicillin G, 50 �g/mL streptomycin sulfate, and 10 IU/mL eCG/ hCG. These IVM oocytes were either activated for parthenogenesis or in vitro-fertilized (IVF). For IVF, oocytes were incubated with 5 � 106/mL of cryopreserved epididymal sperm in PGM-tac medium (Yoshioka et al. 2003 Biol. Reprod. 69, 2092-2099) for 20 h. Embryos were treated for removal of cytoplasmic lipid droplets (delipation; Nagashima et al. 1995 Nature 374, 416) at the 4- to 8-cell stages, around 50 to 54 h after activation or insemination. After culture in NCSU23 for 15 h, they were vitrified by the minimum volume cooling (MVC) method. Embryos were equilibrated with equilibration solution containing 7.5% (v/v) ethylene glycol (EG), 7.5% (v/v) dimethylsulfoxide (DMSO), and 20% (v/v) calf serum for 4 min, followed by exposure to vitrification solution containing 15% EG, 15% DMSO, 0.5 M sucrose, and 20% calf serum. Embryos were then loaded onto a Cryotop (Kitazato Supply Co., Tokyo, Japan) and immediately plunged into liquid nitrogen. Vitrified embryos were examined for viability in vitro and in vivo after warming. Their in vitro developmental competence was compared to that of corresponding control (nonvitrified) embryos. Vitrified 4- to 8-cell stage embryos, both parthenogenetic and IVF, showed developmental competence into blastocysts comparable to that of control embryos (parthenogenetic: 46.8%, 36/77 vs. 51.7%, 31/60; IVF: 40.0%, 30/75 vs. 44.3%, 35/79). Of four surrogate gilts that received a total of 251 vitrified parthenogenetic embryos, three became pregnant and had 20 fetuses (8.0%, 22 to 23 days old). Three surrogates gilts that received 267 vitrified IVF embryos all became pregnant. Of those, the one that received 47 embryos was confirmed to have eight fetuses (17.0%, 22 days old) by autopsy. The other two were examined by ultrasonography at 56 and 95 days of gestation and found to be pregnant. These results suggest that porcine embryos derived from IVM oocytes have a potential to develop into live offspring after delipation and MVC vitrification. This study was supported by PROBRAIN.

242 PROMISING TWO-STEP EVALUATION SYSTEM FOR SELECTING HIGH DEVELOPMENTAL COMPETENCE IN VITRO FERTILIZED EMBRYOS IN CATTLE USING WELL-OF-THE-WELL DISH

2015 ◽  
Vol 27 (1) ◽  
pp. 210
Author(s):  
M. Taniai ◽  
M. Takayama ◽  
O. Dochi ◽  
K. Imai
Keyword(s):  
Evaluation System ◽  
Evaluation Method ◽  
Calf Serum ◽  
Time Lapse ◽  
Developmental Competence ◽  
Bovine Embryo ◽  
Chi Square ◽  
Cell Stage ◽  
Chi Square Test

Bovine IVF embryos are evaluated morphologically using light microscopy just before transfer. However, this evaluation method is subjective, and an objective method with more certainty is needed. Sugimura et al. (PLoS ONE 2012 7, e36627) reported a promising system for selecting healthy IVF bovine embryo by using time-lapse cinematography and 5 prognostic factors. This study was to investigate the efficacy of a 2-step evaluation system of IVF embryos using microscopy for selecting high developmental competence IVF embryos. Cumulus-oocyte complexes (COC) were collected by ovarian follicular aspiration (2 to 5 mm diameter) obtained from a local abattoir. The COC (n = 488) were matured in TCM-199 medium supplemented with 5% calf serum (CS) and 0.02 IU mL–1 of FSH at 38.5°C for 20 h in an atmosphere of 5% CO2 (20 COC 100 µL–1 droplets). After 10 h of gametes co-culture (5.0 × 106 sperm cells mL–1), the presumptive zygotes were cultured in 125 µL of CR1 aa medium supplemented with 5% CS in well of-the-well culture dishes (AS ONE, Japan; 25 zygotes well–1) at 38.5°C in an atmosphere of 5% CO2, 5% O2, and 90% N2 for 9 days. Two-step evaluations of embryos were done at 27 and 55 h post-IVF (hpi). In the first step of evaluation, cleavage patterns at 27 hpi were categorized as mono-cell, 2-cell with even blastomeres and without fragments (normal cleavage), 2-cell with uneven blastomeres, and ≥3 blastomeres. During the second step of evaluation, embryos were classified by their number of blastomeres (2 to 5 cells, 6 to 8 cells, and >8 cells) and the absence or presence of multiple fragments (<20 or >20%) at 55 hpi. The data were analysed by chi-square test. The blastocyst rate (BL%) of embryos cleaved before 27 hpi (56.6%, n = 106) was higher (P < 0.01) than those of embryos cleaved after 27 hpi (37.0%, n = 235). A greater percentage (P < 0.05) of 2-cell embryos with normal cleavage (68.0%, n = 50) developed to blastocysts than from with =3 blastomeres at 27 hpi (40.6%, n = 32). Superior BL% (P < 0.01) was obtained from embryos categorized as 6- to 8-cell stage (58.6%, n = 140) and >8 cell stage (70.6%, n = 25) compared with those embryos at the 2- to 5-cell stage at 55 hpi (26.1%, n = 176). Embryos with no fragments (58.0%, n = 467) had higher BL% (P < 0.01) compared with those with <20% fragments (30.7%, n = 127) and having with >20% fragments (17.5%, n = 25) at 55 hpi. The highest of BL% was observed in embryos showing a normal cleavage to 2-cells with at 27 hpi and having >6 cells with no fragments at 55 hpi (95.2%, n = 21, P < 0.01). These results demonstrate that the 2-step evaluation system at 27 and 55 hpi using microscopy is an effective method for selecting IVF embryos with high developmental competence.

Effects of Removal of the Statoacoustic Ganglion Complex upon the Growing Otocyst

1976 ◽  
Vol 85 (6_suppl) ◽  
pp. 2-32 ◽  
Author(s):  
Thomas R. Van De Water
Keyword(s):  
Organ Culture ◽  
Inner Ear ◽  
Nerve Fibers ◽  
Trophic Effect ◽  
Group A ◽  
Sensory Structures ◽  
Statoacoustic Ganglion ◽  
Group B

An experiment was designed to answer the question as to whether or not the neural elements of the statoacoustic ganglion complex have a trophic effect upon the histodifferentiation of the sensory structures of the embryonic mouse inner ear anlage as it develops in vitro. The embryonic inner ear anlage with associated otic mesenchyme and statoacoustic ganglion complex was excised from 11, 12, and 13-day CBA/C57 mouse embryos. The inner ear explants of each gestational age group were further divided into two groups: the first group “A” (with) statoacoustic ganglion was explanted to the organ culture system without further surgical intervention; the second group “B” (without) statoacoustic ganglion underwent further surgical manipulation during which their statoacoustic ganglion complexes were dissected away prior to explantation to in vitro. The explanted embryonic inner ears were allowed to develop in organ culture until the equivalent of gestation day 21 in vivo was reached for each group; then all cultures were fixed and histologically processed and stained by a nerve fiber stain, in combination with a stain for glucoprotein membranes. Each specimen was code labeled and scored for histodifferentiation of sensory structures. Light microscopic observations confirmed that in group “A” cultures, statoacoustic ganglion neurons and their nerve fibers were present in association with the developed sensory structures; neither ganglion cell neurons nor their nerve fibers were found to be present in the sensory structures that developed in the group “B” organ culture specimens. Quantification revealed no consistent trend of greater occurrence of any sensory structure in the groups of explants analyzed. The presence of such a trend would have signified the probable existence of a trophic effect of the statoacoustic ganglion neural elements upon development of inner ear sensory structures in the group “A” explants of the 11, 12, and 13-day embryo inner ear organ culture specimens when compared to the aganglionic group “B” cultures. Microscopic comparison of the sensory structures and their sensory hair cells that developed in the organ cultures revealed no differences in the quality of the histodifferentiation of either group “A” or group “B” explants. A base to apex pattern of histodifferentiation of the organ of Corti sensory structures, which has been described to occur in vivo, was noted to occur in the in vitro developed cochlear ducts of all of the explanted inner ears without respect to whether neural elements were present (“A”) or absent (“B”) during development. It was concluded from the quantification of histodifferentiation data and the above observation on the pattern of differentiation of Corti's organ that no trophic effect of neural elements of the statoacoustic ganglion complex influencing the histodifferentiation of sensory structures of 11, 12, and 13-gestation day mouse embryo inner ear explants as they differentiate in vitro could be demonstrated.

EVALUATION OF THE EFFECT OF FERROUS SULFATE ON ENAMEL DEMINERALIZATION OF HUMAN DECIDUOUS TEETH: AN IN VITRO STUDY

2017 ◽  
Vol 8 (3) ◽  
pp. 83-89
Author(s):  
Johnny Holanda De Gauw ◽  
Lara Maria Melo Costa ◽  
Rodrigo Neves Silva ◽  
Natanael Barbosa Santos ◽  
Maria Dânia Holanda Tenorio
Keyword(s):  
Ferrous Sulfate ◽  
Polarized Light ◽  
In Vitro Study ◽  
Deciduous Teeth ◽  
Control Group ◽  
Group A ◽  
Ph Cycling ◽  
Group B ◽  
Group D

Objective: This study aimed to evaluate the effect of ferrous sulfate (FS) on demineralized and non-demineralized human deciduous teeth. Additionally, it was evaluated the penetration extent of FS and its remineralizing effect on the enamel of deciduous teeth using Polarized Light Microscopy (PLM). Method: The sample comprised 44 human deciduous teeth. The 44 crowns were divided randomly into four groups: group A (FS after demineralization), group B (FS without demineralization), group C (only demineralization), and group D (control group). FS at 0.45 mol/L-1 was used daily (15 days) and demineralization was done by pH cycling (7 days). Then, three longitudinal slices of the crowns were photographed using PLM. The degree of penetration of the lesion or stain was measured in micrometers, as well as the distance between the external enamel surface and the core of lesion. Results: Group A showed a dark stain on the outer surface of enamel larger than the group B. It is suggested, a remineralizing effect when comparing groups, A and C. The mean depth and standard deviation for groups A, B, and C were 4.27µm (±1.49), 3.72 µm (±1.68) and 5.00 µm (±1.84), respectively. No dark stains were observed in group D. Conclusion: FS stained the demineralized and non-demineralized human deciduous teeth. However, dark stains in the non-demineralized teeth were smaller or absent, than in the demineralized teeth. Therefore, FS may have a protective effect against demineralization.

Developmental competence in vitro and in vivo of cryopreserved, hatched blastocysts from the dromedary camel (Camelus dromedarius)

2004 ◽  
Vol 16 (6) ◽  
pp. 605 ◽  
Author(s):  
J. A. Skidmore ◽  
M. Billah ◽  
N. M. Loskutoff
Keyword(s):  
Survival Rate ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Sodium Lactate ◽  
Developmental Competence ◽  
Penicillin G ◽  
Dromedary Camel ◽  
Dromedary Camels

The present paper describes experiments designed to investigate methods for cryopreserving embryos from dromedary camels. Because preliminary studies had shown ethanediol to be the best cryoprotectant to use for camel embryos, the current experiments were performed to determine the minimum exposure time to 1.5 m ethanediol required to achieve cryoprotection. The uteri of 30 donor camels were flushed non-surgically 8 days after mating. Embryos were recovered and 158 were assigned to one of three groups, which were exposed to 1.5 m ethanediol for either 10 min (n = 67), 5 min (n = 51) or 1 min (n = 40). Embryos were subsequently thawed and rehydrated by expelling either directly into holding medium (HM; HEPES-buffered Tyrode's medium containing sodium lactate and 3 mg mL−1 bovine serum albumin, 10% fetal calf serum, 100 IU mL−1 penicillin G, 100 μg mL−1 streptomycin and 25 μg mL−1 amphotercin B) or initially into HM containing 0.2 m sucrose for 5 or 10 min. The survival rate of all embryos immediately post-thawing, as judged by the morphological appearance of the embryos, was high (91%), but was greatly reduced after 2 h culture (59%). Ninety-two embryos were transferred to recipient camels resulting in 18 viable fetuses (1 min ethanediol exposure, n = 1/15; 5 min ethanediol exposure, n = 3/34; 10 min ethanediol exposure, n = 14/43). Of the embryos rehydrated directly in HM, six of 65 resulted in viable fetuses and those rehydrated initially in 0.2 m sucrose for 5 or 10 min resulted in nine of 47 and three of 46 fetuses respectively. From these experiments, we conclude that camel embryos can be cryopreserved using ethanediol as a cryoprotectant when the embryos are cooled slowly (to 33°C) before being plunged into liquid nitrogen for storage.

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