Efficiency of SCNT Buffalo (<i>Bubalus bubalis</i>) Embryos in Different Culture Medium and Analysis of mRNA Expression of Insulin-like Growth Factors During Embryogenesis

Pandey, Alok; Gupta, S.C.; Singh, Namita; Rana, J. S.; Gupta, Neelam

doi:10.1111/j.1439-0531.2009.01353.x

Cited by 7 publications

(7 citation statements)

References 63 publications

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“…For instance, Han et al (2003) described that bovine SCNT blastocysts have increased expression of IGF2 compared to IVF blastocysts. Furthermore, these authors demonstrated that IGF2 expression was greater in the SCNT embryos cultured in serum containing medium, suggesting that the presence of exogenous proteins in the culture medium aff ected critical development points (Pandey et al, 2010). In agreement with some previous fi ndings, the low expression of IGF2 observed in our bovine SCNT embryos cultured in K-S/BSA medium could be responsible for the low pre-implantation development observed presently.…”

Section: Discussionsupporting

confidence: 91%

Culture medium composition affects the gene expression pattern and in vitro development potential of bovine somatic cell nuclear transfer (SCNT) embryos

2013

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Different culture systems have been studied that support development of somatic cell nuclear transfer (SCNT) embryos up to the blastocyst stage. However, the use of sequential and two-step culture systems has been less studied. The objective of the present study was to examine the developmental potential and quality of bovine SCNT embryos cultured in different two-step culture media based on KSOM, SOF and the macromolecules FBS and BSA (K-K/FBS, K-S/BSA and K-K/BSA, respectively). No differences were observed in the cleavage rate for any of the culture systems. However, there was a significant difference (P<0.01) in the rate of blastocyst development, with the K-K/ FBS culture system yielding a higher rate of blastocysts (28%) compared to other treatments (18 and 15%, for K-S/BSA and K-K/BSA, respectively). Although quality of embryos, as assessed by the total number of cells, was not different, the apoptosis index was significantly affected in the sequential culture system (K-S/BSA). Gene expression analysis showed alterations of DNMT1, IGF2, LIF, and PRDX6 genes in embryos cultured in K-S/FBS and of SOD2 in embryos cultured in K-K/BSA. In conclusion, we demonstrated that culture medium may affect not only the developmental potential of SCNT embryos but also, more importantly, the gene expression pattern and apoptotic index, presenting the possibility to manipulate the culture medium composition to modulate global gene expression and improve the overall efficiency of this technique.

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Section: Discussionsupporting

confidence: 91%

Culture medium composition affects the gene expression pattern and in vitro development potential of bovine somatic cell nuclear transfer (SCNT) embryos

2013

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“…IGF-2R mRNA levels in 2-cell, 4-cell, and hatched blastocysts cultured in serum free-defined medium were higher (P \ 0.05) that for undefined media-containing serum. Culture conditions of embryos, particularly the presence of serum in culture medium, could affect expression of this gene, as observed previously by Fernández-Gonzalez et al (2009) in mouse andPandey et al (2010) in buffalo SCNT. Giraldo et al (2008) reported that abnormal pattern of transcript may be due to epigenetic modifications due to culture conditions, which directly affect chromatin remodelling and is correlated to imprinted genes.…”

Section: Discussionsupporting

confidence: 56%

Differences in developmental competence and gene expression profiles between buffalo (Bubalus bubalis) preimplantation embryos cultured in three different embryo culture media

et al. 2016

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The objective of this study was to compare effects of in vitro culture systems on embryonic development and expression patterns of developmentally important genes in preimplantation buffalo embryos. After IVM/IVF presumptive zygotes were cultured in one of three systems: undefined TCM-199, mCR2aa medium supplemented with 10 % FBS and defined PVA-myo-inositol-phosphate-EGF medium. No (P [ 0.05) differences at 2-cell, 4-cell and 8-cell to 16-cell stages were observed among the three cultured media used, however, increased (P \ 0.05) blastocyst yield, cell number and hatching rate were found in defined medium compared to undefined media. The expression patterns of genes implicated in embryo metabolism (GLUT-1), anti-apoptosis (BCL-2), imprinting (IGF-2R), DNA methylation (DNMT-3A) and maternal recognition of pregnancy (IFNT) were increased (P \ 0.05) in hatched blastocysts derived from defined medium compared to undefined media. In conclusion, serum-free, defined medium improved developmental competence of in vitro cultured buffalo embryos. Whether these differences in morphological development and gene expression have long-term effects on buffalo calves born after embryo transfer remains unknown. However, it is possible that early adaptations of the preimplantation embryo to its environment persist during fetal and post-natal development.

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“…This study is potentially based on the assumption that the expression of genes related to stress response, such as HSPS can also be influenced by inadequate culture conditions (Christians et al. 1997; Pandey et al. 2010).…”

Section: Discussionmentioning

confidence: 99%

“…We chose HSPA-1A as target gene to analyse early bubaline embryonic response to thermal and other intercellular environmental stress in pre-implantation development, because HSPA-1A is synthesized not only in response to stress but also functions as molecular chaperon. This study is potentially based on the assumption that the expression of genes related to stress response, such as HSPS can also be influenced by inadequate culture conditions (Christians et al 1997;Pandey et al 2010). Findings of this study demonstrate the higher level of HSPA-1A in IVP embryos as compared to IVD group, suggesting stress on embryos during in vitro culture environment.…”

Section: Discussionmentioning

confidence: 99%

Expression and Characterization of Constitutive Heat Shock Protein 70.1 (HSPA‐1A) Gene in In Vitro Produced and In Vivo‐Derived Buffalo (Bubalus bubalis) Embryos

Sharma

Nath

Prasad

et al. 2012

Reprod Domestic Animals

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Cells are blessed with a group of stress protector molecules known as heat shock proteins (HSPs), amongst them HSP70, encoded by HSPA-1A gene, is most abundant and highly conserved protein. Variety of stresses hampers the developmental competence of embryos under in vivo and in vitro conditions. Present work was designed to study the quantitative expression of HSPA-1A mRNA in immature oocytes (IMO), matured oocytes (MO), in vitro produced (IVP) and in vivo-derived (IVD) buffalo embryos to assess the level of stress to which embryos are exposed under in vivo and in vitro culture conditions. Further, HSPA-1A gene sequence was analysed to determine its homology with other mammalian sequences. The mRNA expression analysis was carried out on 72 oocytes (40 IMO; 32 MO), 76 IVP and 55 IVD buffalo embryos. Expression of HSPA-1A was found in oocytes and throughout the developmental stages of embryos examined irrespective of the embryo source; however, higher (p < 0.05) expression was observed in 8-16 cell, morula and blastocyst stages of IVP embryos as compared to IVD embryos. Phylogenetic analysis of bubaline HSPA-1A revealed that it shares 91-98% identity with other mammalian sequences. It can be concluded that higher level of HSPA-1A mRNA in IVP embryos in comparison with in vivo-derived embryos is an indicator of cellular stress in IVP system. This study suggests need for further optimization of in vitro culture system in which HSPA-1A gene could be used as a stress biomarker during pre-implantation development.

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Efficiency of SCNT Buffalo (Bubalus bubalis) Embryos in Different Culture Medium and Analysis of mRNA Expression of Insulin-like Growth Factors During Embryogenesis

Cited by 7 publications

References 63 publications

Culture medium composition affects the gene expression pattern and in vitro development potential of bovine somatic cell nuclear transfer (SCNT) embryos

Culture medium composition affects the gene expression pattern and in vitro development potential of bovine somatic cell nuclear transfer (SCNT) embryos

Differences in developmental competence and gene expression profiles between buffalo (Bubalus bubalis) preimplantation embryos cultured in three different embryo culture media

Expression and Characterization of Constitutive Heat Shock Protein 70.1 (HSPA‐1A) Gene in In Vitro Produced and In Vivo‐Derived Buffalo (Bubalus bubalis) Embryos

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