Cells under pressure: how sublethal hydrostatic pressure stress treatment increases gametes' and embryos' performance

2011 ◽  
Vol 23 (1) ◽  
pp. 48 ◽  
Author(s):  
Csaba Pribenszky ◽  
Gabor Vajta
Keyword(s):  
Hydrostatic Pressure ◽  
Cell Survival ◽  
New Approach ◽  
In Vitro Development ◽  
Stress Treatment ◽  
Sublethal Stress ◽  
Minimal Damage ◽  
Pressure Stress ◽  
Vitro Development

The principal approach in in vitro embryo culture and manipulation has been a defensive one: procedures aim to satisfy passively the supposed or real physiological needs of gametes and embryos. Similarly, during cryopreservation the aim is to cause minimal damage to cells whilst attempting to obtain the highest achievable cell survival. However, carefully chosen and precisely controlled sublethal stress treatment of cells has been described to improve embryos’ and gametes’ performance, and, as a consequence, subsequent morphological survival, fertilisation, in vitro development, pregnancy and farrowing rates improved considerably compared with untreated controls. This review summarises studies that open up a new approach: instead of – and besides – trying to passively reduce the harm to cells during in vitro manipulations and culture, procedures may also prepare the cells themselves to ward off or reduce the damage by turning up the cells’ own, inner capacities.

Vitrification of bovine blastocysts pretreated with sublethal hydrostatic pressure stress: evaluation of post-thaw in vitro development and gene expression

2011 ◽  
Vol 23 (4) ◽  
pp. 585 ◽  
Author(s):  
E. Siqueira Filho ◽  
E. S. Caixeta ◽  
C. Pribenszky ◽  
M. Molnar ◽  
A. Horvath ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hydrostatic Pressure ◽  
Transcript Abundance ◽  
Developmental Competence ◽  
Hatching Rate ◽  
In Vitro Development ◽  
Stress Treatment ◽  
Pressure Stress ◽  
Vitro Development

Sublethal stress treatment has been reported to enhance gametes’ performance in subsequent procedures, such as cryopreservation. The aim of the present study was to evaluate the effect of different equilibration times between the termination of a sublethal hydrostatic pressure (HP) stress treatment and the initiation of vitrification on the post-thaw survival, continued in vitro development, hatching rate and gene expression of selected candidate genes of in vitro-produced (IVP) expanded bovine blastocysts. Day 7 IVP blastocysts were subjected to 600 bar pressure for 60 min at 32°C. Immediately after pressure treatment (HP0h) or after 1 or 2 h incubation (HP1h and HP2h groups, respectively), embryos were either vitrified and warmed using the open pulled straw method, followed by 72 h in vitro culture or were stored at –80°C until gene expression analysis. Re-expansion and hatching rates after vitrification–warming were significantly (P < 0.05) higher in the HP0h (88 and 76%, respectively) and HP1h (90 and 75%, respectively) groups than in the untreated (82 and 63%, respectively) and HP2h groups (79 and 70%, respectively). Moreover, the HP1h group showed further improvement in the speed of re-expansion and resumption of normal in vitro development. Cumulative analysis of all genes (SC4MOL, HSP1A1A, SOD2 and GPX4) revealed a similar pattern of expression, with a tendency for peak transcript abundance 1 h after HP treatment. Application of HP stress treatment was found to be efficient in increasing the in vitro developmental competence of vitrified bovine embryos.

89 IMPROVED POST-WARMING DEVELOPMENTAL COMPETENCE OF OPEN PULLED STRAW-VITRIFIED IN VITRO-PRODUCED BOVINE BLASTOCYSTS BY SUBLETHAL HYDROSTATIC PRESSURE PRETREATMENT

2008 ◽  
Vol 20 (1) ◽  
pp. 125 ◽  
Author(s):  
C. Pribenszky ◽  
F. E. Siqueira ◽  
M. Molnár ◽  
A. Harnos ◽  
R. Rumpf
Keyword(s):  
Hydrostatic Pressure ◽  
Developmental Competence ◽  
Equilibration Time ◽  
In Vitro Development ◽  
Custom Made ◽  
Boar Semen ◽  
Pre Treatment ◽  
Vitro Development ◽  
Hatching Rates

Vitrification of in vitro (IVP)-produced bovine blastocysts is well established, reaching post-warming hatching rates close to 70–80% in vitro. Nevertheless, improvements still are needed regarding realizable pregnancy rates. High hydrostatic pressure (HHP) treatment of fresh boar semen before freezing increased the litter size achieved by insemination of frozen–thawed boar semen (Kuo et al. 2007 6th Int. Conf. Boar Semen Pres, Alliston, ON, Canada, poster #22); HHP treatment-related improvements were also observed in the in vitro cryosurvival of mouse blastocysts (Pribenszky et al. 2004 Reprod. Fertil. Dev. 17, 199–200), bull and boar semen, and pig oocytes, theoretically by the sublethal stress-induced production/stabilization of shock proteins (Pribenszky et al. 2006 Reprod. Fertil. Dev. 18, 162–163; 2007 Reprod. Fertil. Dev. 19, 181–182; and b; Du et al. 2007). The aim of the present study was to improve the post-warming in vitro developmental competence of vitrified bovine IVP blastocysts through the application of HHP as pre-treatment, in order to apply the protocol in later in vivo experiments. Day 7 IVP blastocysts were aspirated in TQC holding medium (AB Technology, Sao Paulo, Brazil) into 0.25-mL straws. Straws were pressure-treated in a custom-made hydrostatic pressure chamber (Cryo-Innovation Ltd., Budapest, Hungary), using water as pressure medium. Six hundred bar pressure was applied for 60 min at 32�C. Immediately after pressure treatment, or following 60- or 120-min incubation, embryos were vitrified and warmed using open pulled straws (OPS) according to the method of Vajta et al. (1998 Mol. Reprod. Dev. 51, 53–58). Untreated blastocysts were vitrified as controls. After warming, embryos were cultured in vitro in SOF (Holm et al. 1999 Theriogenology 52, 683–700) for 72 h. Embryos were checked for re-expansion and hatching at 4, 24, 48, and 72 h post-warming. For the experiment, 404 blastocysts were used in 5 replicates. Logistic regression was used for statistical evaluation. All vitrified groups were inferior compared to the non-vitrified control (97%, 97%, 98, and 100% expansion; 0%, 23%, 72 and 91% hatching at 4, 24, 48, and 72 h, respectively). HHP treatment had a significant effect (P > 0.05) on the post-warming developmental competence of vitrified blastocysts. HHP treatment followed by 60 min of equilibration proved to be superior among all treatment groups regarding both re-expansion and hatching rates and the speed of resumption of normal in vitro development (HHP treatment followed by a 60-min equilibration time before vitrification/warming: re-expansion rates: 88%, 89%, 85, and 90%; hatching rates: 0%, 22%, 51, and 73% v. non-treated vitrified/warmed controls: re-expansion rates: 63%, 69%, 71, and 81%; hatching rates: 0%, 6%, 43, and 63%; at 4, 24, 48, and 72 h post-warm, respectively). In conclusion, hydrostatic pressure pre-treatment significantly improved in vitro survival and hatching rates as well as the speed of resumption of normal in vitro development. Further studies are needed to reveal the molecular-biological implications of the HHP treatments, as well as field trials to test if the in vitro improvements can be confirmed by pregnancy and birth rates. This work was supported by EMBRAPA and a Kozma grant, Hungary.

scholarly journals In vitro development of preimplantation porcine embryos using alginate hydrogels as a three-dimensional extracellular matrix

2014 ◽  
Vol 26 (7) ◽  
pp. 943 ◽  
Author(s):  
Catherine N. Sargus-Patino ◽  
Elane C. Wright ◽  
Sarah A. Plautz ◽  
Jeremy R. Miles ◽  
Jeff L. Vallet ◽  
...  
Keyword(s):  
Cell Survival ◽  
Culture System ◽  
Culture Media ◽  
Morphological Changes ◽  
Three Dimensional ◽  
In Vitro Development ◽  
Alginate Hydrogels ◽  
Vitro Development

Between Days 10 and 12 of gestation, porcine embryos undergo a dramatic morphological change, known as elongation, with a corresponding increase in oestrogen production that triggers maternal recognition of pregnancy. Elongation deficiencies contribute to embryonic loss, but exact mechanisms of elongation are poorly understood due to the lack of an effective in vitro culture system. Our objective was to use alginate hydrogels as three-dimensional scaffolds that can mechanically support the in vitro development of preimplantation porcine embryos. White cross-bred gilts were bred at oestrus (Day 0) to Duroc boars and embryos were recovered on Days 9, 10 or 11 of gestation. Spherical embryos were randomly assigned to be encapsulated within double-layered 0.7% alginate beads or remain as non-encapsulated controls (ENC and CONT treatment groups, respectively) and were cultured for 96 h. Every 24 h, half the medium was replaced with fresh medium and an image of each embryo was recorded. At the termination of culture, embryo images were used to assess morphological changes and cell survival. 17β-Oestradiol levels were measured in the removed media by radioimmunoassay. Real-time polymerase chain reaction was used to analyse steroidogenic transcript expression at 96 h in ENC and CONT embryos, as well as in vivo-developed control embryos (i.e. spherical, ovoid and tubular). Although no differences in cell survival were observed, 32% (P < 0.001) of the surviving ENC embryos underwent morphological changes characterised by tubal formation with subsequent flattening, whereas none of the CONT embryos exhibited morphological changes. Expression of steroidogenic transcripts STAR, CYP11A1 and CYP19A1 was greater (P < 0.07) in ENC embryos with morphological changes (ENC+) compared with CONT embryos and ENC embryos with no morphological changes (ENC–), and was more similar to expression of later-stage in vivo-developed controls. Furthermore, a time-dependent increase (P < 0.001) in 17β-oestradiol was observed in culture media from ENC+ compared with ENC– and CONT embryos. These results illustrate that preimplantation pig embryos encapsulated in alginate hydrogels can undergo morphological changes with increased expression of steroidogenic transcripts and oestrogen production, consistent with in vivo-developed embryos. This alginate culture system can serve as a tool for evaluating specific mechanisms of embryo elongation that could be targeted to improve pregnancy outcomes.

In vitro development of human triploid zygotes reconstructed by pronuclear transfer

2002 ◽  
Vol 78 ◽  
pp. S180-S181
Author(s):  
John Zhang ◽  
Yi Ming Shu ◽  
Lewis C Krey ◽  
Hui Liu ◽  
Guang Lun Zhuang ◽  
...  
Keyword(s):  
In Vitro Development ◽  
Pronuclear Transfer ◽  
Vitro Development

Gallic acid promotes the in vitro development of sheep secondary isolated follicles involving the phosphatidylinositol 3-kinase pathway

2021 ◽  
pp. 106767
Author(s):  
Gizele A.L. Silva ◽  
Luana B. Araújo ◽  
Larissa C.R. Silva ◽  
Bruna B. Gouveia ◽  
Ricássio S. Barberino ◽  
...  
Keyword(s):  
Gallic Acid ◽  
Phosphatidylinositol 3 Kinase ◽  
In Vitro Development ◽  
Vitro Development ◽  
Kinase Pathway ◽  
Phosphatidylinositol 3

scholarly journals Necrostatin-1 Supplementation to Islet Tissue Culture Enhances the In-Vitro Development and Graft Function of Young Porcine Islets

2021 ◽  
Vol 22 (16) ◽  
pp. 8367
Author(s):  
Hien Lau ◽  
Shiri Li ◽  
Nicole Corrales ◽  
Samuel Rodriguez ◽  
Mohammadreza Mohammadi ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Oxygen Consumption ◽  
Insulin Secretion ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
In Vitro Development ◽  
Porcine Islets ◽  
Vitro Development

Pre-weaned porcine islets (PPIs) represent an unlimited source for islet transplantation but are functionally immature. We previously showed that necrostatin-1 (Nec-1) immediately after islet isolation enhanced the in vitro development of PPIs. Here, we examined the impact of Nec-1 on the in vivo function of PPIs after transplantation in diabetic mice. PPIs were isolated from pancreata of 8–15-day-old, pre-weaned pigs and cultured in media alone, or supplemented with Nec-1 (100 µM) on day 0 or on day 3 of culture (n = 5 for each group). On day 7, islet recovery, viability, oxygen consumption rate, insulin content, cellular composition, insulin secretion capacity, and transplant outcomes were evaluated. While islet viability and oxygen consumption rate remained high throughout 7-day tissue culture, Nec-1 supplementation on day 3 significantly improved islet recovery, insulin content, endocrine composition, GLUT2 expression, differentiation potential, proliferation capacity of endocrine cells, and insulin secretion. Adding Nec-1 on day 3 of tissue culture enhanced the islet recovery, proportion of delta cells, beta-cell differentiation and proliferation, and stimulation index. In vivo, this leads to shorter times to normoglycemia, better glycemic control, and higher circulating insulin. Our findings identify the novel time-dependent effects of Nec-1 supplementation on porcine islet quantity and quality prior to transplantation.

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