Bioactive peptide analogs: In vivo and in vitro production

1990 ◽  
pp. 287-317
Author(s):  
Horst Kleinkauf ◽  
Hans von Doehren
Keyword(s):  
Bioactive Peptide ◽  
In Vitro Production ◽  
Peptide Analogs ◽  
Vitro Production

257 DIFFERENCES BETWEEN BOS TAURUS AND BOS INDICUS EMBRYOS: TRANSCRIPTS AMOUNTS

2010 ◽  
Vol 22 (1) ◽  
pp. 285
Author(s):  
S. Wohlres-Viana ◽  
M. M. Pereira ◽  
A. P. Oliveira ◽  
J. H. M. Viana ◽  
M. A. Machado ◽  
...  
Keyword(s):  
Production System ◽  
Production Systems ◽  
Bos Taurus ◽  
Bos Indicus ◽  
In Vitro Production ◽  
Embryo Production ◽  
Peroxiredoxin 1 ◽  
Vitro Production

The Zebu breeds (Bos indicus) are different from European breeds (Bos taurus) in some aspects of their reproductive physiology, including follicle recruitment, number of follicular waves, and oocyte ultrastructure. On the other hand, embryos produced in vivo and in vitro show morphological and developmental differences, which can be related to culture environment. The aim of this study was to evaluate the effect of breed (Gyr v. Holstein) within embryo production system (in vivo and in vitro), as well as effect of production systems within breeds on relative abundance of transcripts related to formation, survival, and subsequent development of blastocysts, such as those involved in water and small solutes transport (Aquaporins 3 and 11), blastocoel formation (Na+/K+-ATPase a1 and |52), and cellular stress response (Peroxiredoxin 1). For in vivo embryo production, donors were superstimulated with FSH and inseminated, and embryos were recovered 7 days after AI. For in vitro embryo production, oocytes recovered by ovum pickup were in vitro matured and fertilized and then cultured for 7 days in culture medium under 5% CO2 at 38.5°C. For each group, blastocysts (n = 15) distributed in 3 pools were used for RNA extraction (RNeasy MicroKit, Qiagen, Valencia, CA, USA), followed by RNA amplification (Messageamp II amplification kit, Ambion-Applied Biosystems, Foster City, CA, USA) and reverse transcription (SuperScript III First-Stand Synthesis Supermix, Invitrogen, Carlsbad, CA, USA). The cDNA were submitted to real-time PCR, using the H2a gene as endogenous control, and analyzed by REST© software. To evaluate breed effect within the production systems, 2 comparisons were performed: (1) in vivo: Gyr v. Holstein and (2) in vitro: Gyr v. Holstein, considering Holstein data as 1.00. To evaluate production system effect within breeds, 2 comparisons were performed: (1) Gyr: in vivo v. in vitro and (2) Holstein: in vivo v. in vitro, considering in vivo produced embryo data as 1.00. The results are shown as mean ± SEM. For in vivo comparison between breeds, Aquaporin 3 (1.66 ± 0.77), Na+/K+-ATPase a1 (1.61 ± 0.56), and Peroxiredoxin 1 (1.61 ± 0.66) were up-regulated (P < 0.05) in Gyr embryos when compared with Holstein embryos, whereas for in vitro comparison, no differences (P > 0.05) were found. For comparisons between production systems within breeds, only Peroxiredoxin 1 (0.31 ± 0.39) was down-regulated (P < 0.01) in in vitro produced Gyr embryos when compared with in vivo counterparts. No differences (P > 0.05) were found between production systems for the Holstein breed. In conclusion, these data suggest that there is a difference on gene expression between Bos taurus and Bos indicus blastocysts, but such difference between breeds can be attenuated by the in vitro production system, indicating an embryo adaptation to the in vitro culture conditions. The data also suggest that the in vitro production system can influence the amount of transcripts in Gyr embryos. Other genes should be evaluated for a better understanding of these differences. Financial support was provided by CNPq and FAPEMIG.

scholarly journals Reprogrammed Cell?based Therapy for Liver Disease: From Lab to Clinic

2017 ◽  
Vol 1 (1) ◽  
pp. 20-28 ◽  
Author(s):  
Amir Mehdizadeh ◽  
Masoud Darabi
Keyword(s):  
Cellular Reprogramming ◽  
Gene Correction ◽  
In Vitro Production ◽  
Efficient Treatment ◽  
Cell Based Therapy ◽  
Number Of Patients ◽  
End Stage ◽  
Vitro Production

A large number of patients are affected by liver dysfunction worldwide. Liver transplantation is the only efficient treatment in a variety of enduring liver disorders including inherent and end-stage liver diseases. The generation of human functional hepatocytes in high quantities for liver cell therapy is an important goal for ongoing therapies in regenerative medicine. Reprogrammed cells are considered as a promising and unlimited source of hepatocytes, mainly because of their expected lack of immunogenicity and minimized ethical concerns in clinical applications. Despite gained advances in the reprogramming of somatic cells to functional hepatocytes in vitro, production of primary adult hepatocytes that can proliferate in vivo still remains inaccessible. As part of efforts toward translation of cell reprogramming science into clinical practice, more careful cell selection strategies should be integrated into improvement of dedifferentiation and redifferentiation protocols, especially in precision medicine where gene correction is needed. Furthermore, advances in cellular reprogramming highlight the need for developing and evaluating novel standards addressing clinical research interests in this field.

86 Comparison of in vivo and in vitro embryo production in Bonsmara donors

2019 ◽  
Vol 31 (1) ◽  
pp. 168
Author(s):  
B. H. Bernal ◽  
J. L. Barajas ◽  
J. A. Ortega ◽  
A. Cedeño ◽  
S. Andrada ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Gonadotropin Releasing Hormone ◽  
P Value ◽  
In Vitro Production ◽  
Embryo Production ◽  
Releasing Hormone ◽  
Gradient Technique ◽  
Vitro Production

A retrospective analysis of embryo production records from 2013 to 2017 was carried out to evaluate the in vivo and in vitro production (IVP) of embryos in donors of the Bonsmara breed (i.e. tropically adapted Bos taurus). Only donors with production records of both in vivo and in vitro embryos during the same period were used. A total of 127 superovulations and ova/embryo collections of 19 donors were evaluated. The donors were superstimulated with the following protocol: on Day 0 they received a device with 1g of progesterone (DIB, Zoetis, Argentina), 50mg of rogesterone (Progestar, Zoetis), and 5mg of oestradiol-17β (17ßOestradiol, Rio de Janeiro, Argentina) or 2mg of oestradiol benzoate (Gonadiol, Zoetis) intramuscularly (IM) at the same time. Superstimulatory treatments were initiated on the morning of Day 4 with Folltropin-V (Vetoquinol, France; total dose=240 to 340mg IM) in twice-daily decreasing doses over 4 days. All donors received 2 IM injections of 500µg of cloprostenol (Ciclase DL, Zoetis) on the morning and afternoon of Day 6 and; the intravaginal devices were removed on the morning of Day 7 and 100µg of Gonadorelin (gonadotropin-releasing hormone, Gonasyn gdr, Zoetis) was given on the morning of Day 8. Donors were inseminated using semen from 9 Bonsmara bulls, 12 and 24h after gonadotropin-releasing hormone. On Day 15, ova/embryos were collected and classified according to IETS standards. A total of 89 follicular aspirations (ovum pickup) of 19 donors for IVP were evaluated. The ovum pickups were performed at random stages of oestrous cycle, without superstimulation or other hormone treatments. A total of 1109 viable oocytes (12.5±0.9 per ovum pickup) were collected and matured for 24h in 100-µL drops of maturation medium (TCM-199, supplemented with hormones) under mineral oil and incubated at 38.5°C in 5.5% CO2 and humidity at saturation. Fertilization was performed using 3 Bonsmara bulls that were also used for in vivo embryo production. Viable sperm were obtained using the percoll gradient technique (45-90%). The sperm pellet was dissolved in TL-Sperm, centrifuged, and then diluted to a final concentration of 1.5×106 sperm/mL. Zygotes were stripped and placed in drops of 100µL of SOF medium supplemented with 0.4% BSA under oil at 38.8°C, 5.5% CO2, 7% O2, and humidity at saturation for 7 days. The culture medium was renewed on Days 3 and 5. The data were analysed using the GLM procedure of SAS (SAS Institute Inc., Cary, NC, USA), a P-value &lt;0.05 was considered significant. The mean (±standard error of the means) number of CL, ova/embryos collected, fertilized ova, and transferable embryos were 12.9±0.6, 8.8±0.6, 6.6±0.5, and 4.7±0.4, respectively. A total of 662 oocytes (66.3±2.4%) cleaved 48h post-IVF. On Day 7, an average of 4.4±0.3 embryos were produced. No differences were detected in the number of transferable embryos produced in vivo v. those produced in vitro. Furthermore, no significant differences were found between the techniques or bulls on the proportion of embryos produced in relation to the ova/embryos or oocytes obtained (in vivo 51.5±3.2% v. in vitro 42.9±2.5%). In conclusion, the in vivo and in vitro production of embryos are both effective alternatives to increase the number of offspring from valuable Bonsmara donors.

​Effect of Season on Quantity and Competence of Oocytes Recovered Transvaginally from Holstein Cows for In vitro Fertilization

2021 ◽  
Author(s):  
H.Z. Guerrero-Gallego ◽  
G. Calderon-Leyva ◽  
O. Angel-Garcia ◽  
J.M. Guillen-Muñoz ◽  
C. Leyva ◽  
...  
Keyword(s):  
Holstein Cows ◽  
In Vitro Production ◽  
Vitro Fertilization ◽  
Follicular Aspiration ◽  
Four Seasons ◽  
Aspiration Technique ◽  
Vitro Production ◽  
Intense Heat

Background: Season of the year can affect the reproductive behavior in Holstein cows, altering the competition of the oocytes, reflecting a reduced production of embryos. The objective of this study was to evaluate the average of total oocytes, competition of oocytes and embryos in the in vitro production process at different season of the year in Holstein cows. Methods: During the four seasons of the year, was performed on each of the oocyte donor cows (winter, n = 957; spring, n = 1571; summer, n = 1776; autumn, n = 1128), by in vivo transvaginal follicular aspiration technique after the collection were subjected to the embryos production in vitro. Result: The highest number of total embryos were produced in winter and autumn, compared to spring and summer (3.76±0.16 and 3.54±0.18 vs. 2.73±0.11 and 2.45±0.10; respectively, P less than 0.05). During winter, a higher percentage of oocyte competition was observed, followed by autumn and spring and less competition shown in summer (26.03±0.39, 19.08±0.29, respectively, P less than 0.05). The quantity and competence of the oocytes collected and in vitro embryo production were drastically reduced during the hottest months of the year in this area of intense heat.

In Vivo and In Vitro Production Options for Fungal Secondary Metabolites

2008 ◽  
Vol 5 (2) ◽  
pp. 234-242 ◽  
Author(s):  
Patrick Schneider ◽  
Mathias Misiek ◽  
Dirk Hoffmeister
Keyword(s):  
Secondary Metabolites ◽  
In Vitro Production ◽  
Vitro Production ◽  
Fungal Secondary Metabolites

scholarly journals 35 PRODUCTION OF CLONED MINIATURE CALVES USING CYTOPLASTS FROM COWS OF STANDARD SIZE

2005 ◽  
Vol 17 (2) ◽  
pp. 167
Author(s):  
M. Givens ◽  
R. Carson ◽  
K. Riddell ◽  
D. Stringfellow ◽  
J. Waldrop ◽  
...  
Keyword(s):  
Poor Response ◽  
Fetal Anomalies ◽  
In Vitro Production ◽  
Standard Size ◽  
Fetal Wastage ◽  
Chromatin Transfer ◽  
Vitro Production ◽  
Human And Animal Diseases

Numerous genetically similar cattle for use in research or teaching can be produced with chromatin transfer technology (Sullivan et al. 2004 Biol. Reprod. 70, 146–153). Miniature cattle can provide advantageous biotechnological models for teaching and study of human and animal diseases. Miniature cattle are approximately one fourth the size of standard cattle and, therefore, represent a potentially less expensive, safer, and more easily managed animal model. Our limited attempts to reproduce miniature cattle via in vivo and in vitro production of embryos resulted in poor response to superovulation and logistical challenges in recovering embryos or oocytes due to the small size of the animals. Thus, the purpose of this study was to evaluate the potential of embryos derived by chromatin transfer from fibroblasts of a miniature cow and cytoplasts from cows of standard size to produce viable offspring after transfer into recipient cows of standard size. The donor of somatic cells was a heifer that weighed 7.7 kg at birth. Chromatin transfer resulted in 19% (82/428) blastocyst formation. A total of 66 cloned blastocysts (65 excellent/good quality, 1 fair quality) were transferred into 26 synchronized recipients. While ultrasound revealed 13 pregnancies prior to 67 days of gestation, only 4 pregnancies of 5 fetuses were maintained beyond 100 days. Parturition was induced with dexamethasone and prostaglandin on Day 286 of gestation. One singleton (12.3 kg) and a set of twins (10.2 and 11.1 kg) were healthy at birth and normal at 1 week of age. Two fully developed singletons, weighing 21.4 and 13.6 kg, died in utero. The latter fetus exhibited a fixed dorsolateral deviation of the neck that complicated delivery despite a caudal obstetrical presentation. No abnormalities were noted in the size or structure of any placenta. Our results indicate that healthy miniature calves can be gestated by recipient cows of standard size after transfer of embryos derived by chromatin transfer. Unfortunately, the fetal wastage, fetal anomalies, and stillbirths observed with standard sized cattle also may occur.

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