Assoc.Prof. Rangsun Parnpai

Year	Title	Journal
2013	Ovarian follicular dynamics, ovarian follicular growth, oocyte yield, in vitro embryo production and repeated oocyte pick up in thai native heifers undergoing superstimulation Chasombat J., Nagai T., Parnpai R., Vongpralub T.	Asian-Australasian Journal of Animal Sciences
Abstract: The objective of this study was to compare the effectiveness of the protocols for superstimulation of follicular growth in Thai native heifers. Heifers (n = 20) were randomly divided into four groups of five heifers/group. Heifers were given a single dose by i.m. administration of 100 mg Follicle Stimulating Hormone dissolved in polyvinylpyrrolidone (FSHp) at 24 h. Ovum pick up (OPU) occurred at 72 h (F24O72 protocol; Group 1) or 96 h (F24O96 protocol; Group 2), and at 36 h and OPU at 72 h (F36O72 protocol; Group 3) or 96 h (F36O 96 protocol; Group 4) after follicular ablation. The dynamics of ovarian follicular growth were monitored by twice-daily ultrasonographic examinations. Blood sample collections were performed every 12 h after initiation of treatment for assessment of FSH, E2 and P4 profiles. All heifers were subjected to eight repeated sequential sessions of OPU. The follicular deviation commenced 24±5.32 h after follicular ablation in all groups. The circulatory FSH surged quickly from 24 to 36 h (>0.8 ng/ml) after follicular ablation and circulatory estrogen levels steadily increased from 36 h until OPU in all groups. At the end of the OPU sessions, the mean number of aspirated follicles/heifer/session in F36O72 protocol (Group 3) and F36O96 protocol (Group 4) were higher than in the two other groups (p<0.05). The number of cumulus-oocyte complexes (COCs), cleaved and day 8 blastocysts rates in the F36O72 protocol (Group 3) were higher than in the other groups (p<0.05). It can be concluded that a single dose i.m. administration of 100 mg FSHp at 36 h and OPU at 72 h after follicular ablation (F36O72 protocol; Group 3) was the most effective protocol for superstimulation of follicular growth for repeated OPU and subsequent in vitro embryo production in Thai native heifers. Copyright © 2013 by Asian-Australasian Journal of Animal Sciences. Keyword: Cattle; COCS; Follicle; FSH; IVP; OPU Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84878528984&doi=10.5713%2fajas.2012.12519&partnerID=40&md5=0564ba455b12d86b15df5dacfbb5b825 DOI: 10.5713/ajas.2012.12519
2012	Discrimination of functional hepatocytes derived from mesenchymal stem cells using FTIR microspectroscopy Ye D., Tanthanuch W., Thumanu K., Sangmalee A., Parnpai R., Heraud P.	Analyst
Abstract: Functional hepatocytes differentiated in vitro from mesenchymal stem cells (MSCs) need to be fully characterized before they could be applied as a therapy to treat liver disease. Here, we employed Fourier Transform Infrared (FTIR) microspectroscopy to investigate the characteristics of hepatocyte-like cells derived from rat bone marrow mesenchymal stem cells (rBM-MSCs) by detecting changes in macromolecular composition occurring during the hepatogenesis process. Partial Least Squares Discriminant Analysis (PLS-DA) enabled us to discriminate undifferentiated rBM-MSCs, and early, mid-stage and late stage rBM-MSCs derived hepatocytes by their characteristic FTIR "spectroscopic signatures". The predominant spectroscopic changes responsible for this discrimination were changes in FTIR absorbance bands at: 3012 cm-1 (cis CC stretch from unsaturated lipids), 2952 cm-1 (νasCH3 from lipids), 2854 cm-1 (νsCH2 from lipids) and 1722 cm-1 (CO stretching from lipids), which were associated with triglyceride and unsaturated fatty acid accumulation in the hepatocyte-like cells occurring during differentiation. Based on these findings, rBM-MSCs derived hepatocytes are characterized by high lipid content which facilitates a means of identifying hepatocytes from their stem cells progenitors by using FTIR microspectroscopy. Other complex changes in spectral bands assigned to proteins and nucleic acids were observed during hepatocyte differentiation indicating that mRNA translation was taking place producing proteins related to the formation of the new hepatocyte-like phenotype, which was corroborated by immunohistochemistry. The results show FTIR microspectroscopy combined with bioinformatic modeling constitutes a powerful new phenotypic-based methodology for monitoring and characterization of the process of stem cell differentiation leading to the formation of hepatocytes, providing complementary information to existing methodologies such as immunohistochemistry and gene analysis, but having advantages of being reagent-free and non-destructive of the sample. © 2012 The Royal Society of Chemistry. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84866521665&doi=10.1039%2fc2an35329f&partnerID=40&md5=e007f923e39fb232ad66cd05eef63b27 DOI: 10.1039/c2an35329f
2012	Effects of vitrification cryoprotectant treatment and cooling method on the viability and development of buffalo oocytes after intracytoplasmic sperm injection Liang Y.Y., Srirattana K., Phermthai T., Somfai T., Nagai T., Parnpai R.	Cryobiology
Abstract: In vitro matured (IVM) buffalo oocytes at the metaphase of the second meiotic division (MII) were vitrified in 20% Me2SO: 20% EG (v/v) and 0.5M sucrose (VA), or 35% EG (v/v), 50mg/mL polyvinylpyrrolidone (PVP), and 0.4M trehalose (VB), either on cryotops or as 2μL microdrops. The viability was assessed after warming by fluorescein diacetate (FDA) staining and all surviving oocytes were subjected to ICSI and ethanol activation. All vitrified groups had similar recovery rates but both VA groups had significantly higher survival and pronuclear formation rates than either of the VB groups. Non treated control oocytes and non cryopreserved oocytes exposed to FDA had significantly higher survival, 2nd polar body extrusion, PN and blastocyst formation rates than any of the four vitrified groups (P<0.05). In conclusion The cryotop and microdrop methods are equally effective for buffalo oocyte vitrification, and although vitrification in VA solution yielded higher rates of survival and formation of 2 pronuclei than VB, the rate of blastocyst formation was comparable for both solutions. A detailed analysis of oocytes that extruded the second polar body after ICSI and activation revealed that only a minority (7-20% of the vitrified and 46-48% of the control oocytes) also had two pronuclei, indicating that normal activation is compromised by vitrification. © 2012 Elsevier Inc. Keyword: Buffalo oocyte; Cryotop; In vitro fertilization; Microdrop; Vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84864316411&doi=10.1016%2fj.cryobiol.2012.04.006&partnerID=40&md5=88bb3af6c4475abda9c01afd7c437d39 DOI: 10.1016/j.cryobiol.2012.04.006
2012	Cryopreservation of immature buffalo oocytes: Effects of cytochalasin B pretreatment on the efficiency of cryotop and solid surface vitrification methods Liang Y., Rakwongrit D., Phermthai T., Somfai T., Nagai T., Parnpai R.	Animal Science Journal
Abstract: The aim of the present study was to compare the efficiency of the solid surface (SSV), cryotop (CT) vitrification methods and cytochalasin B (CB) pretreatment for cryopreservation of immature buffalo oocytes. Cumulus-oocyte complexes (COCs) were placed for 1min in TCM199 containing 10% dimethylsulfoxide (DMSO), 10% ethylene glycol (EG), and 20% fetal bovine serum, and then transferred for 30s to base medium containing 20% DMSO, 20% EG and 0.5mol/L sucrose. CB pretreated ((+)CB) or non-pretreated ((-)CB) COCs were vitrified either by SSV or CT. Surviving vitrified COCs were selected for in vitro maturation (IVM) and in vitro fertilization (IVF). The rate of viable oocytes after vitrification in CT groups (82%) was significantly lower (P<0.05) than that in a fresh control group (100%), but significantly higher (P<0.05) than those in SSV groups (71-72%). Among vitrified groups, the highest maturation rate was obtained in the CT (-)CB group (32%). After IVF, the cleavage and blastocyst formation rates were similar among vitrified groups but significantly lower than those of the control group. In conclusion, a higher survival rate of oocytes after vitrification and IVM was obtained in the CT group compared with that in the SSV group, indicating the superiority of the CT method. Pretreatment with CB did not increase the viability, maturation or embryo development of vitrified oocytes. © 2012 The Authors. Animal Science Journal © 2012 Japanese Society of Animal Science. Keyword: Buffalo oocyte; Cryotop; Cytochalasin B; Solid surface vitrification; Vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84865730594&doi=10.1111%2fj.1740-0929.2012.01013.x&partnerID=40&md5=278c61724ad187e770559da86ec968de DOI: 10.1111/j.1740-0929.2012.01013.x
2012	Segregation of donor cell mitochondrial DNA in gaur-bovine interspecies somatic cell nuclear transfer embryos, fetuses and an offspring Imsoonthornruksa S., Srirattana K., Phewsoi W., Tunwattana W., Parnpai R., Ketudat-Cairns M.	Mitochondrion
Abstract: The fate of foreign mitochondrial DNA (mtDNA) following somatic cell nuclear transfer (SCNT) is still controversial. In this study, we examined the transmission of the heteroplasmic mtDNA of gaur donor cells and recipient bovine oocytes to an offspring and aborted and mummified fetuses at various levels during the development of gaur-bovine interspecies SCNT (iSCNT) embryos. High levels of the donor cell mtDNA were found in various tissue samples but they did not have any beneficial effect to the survival of iSCNT offspring. However, the factors on mtDNA inheritance are unique for each iSCNT experiment and depend on the recipient oocyte and donor cell used, which might play an important role in the efficiency of iSCNT. © 2012 Elsevier B.V. and Mitochondria Research Society. Keyword: Bovine cytoplasm; Endangered species; Gaur; Heteroplasmy; Mitochondrial DNA Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84864770470&doi=10.1016%2fj.mito.2012.07.108&partnerID=40&md5=ad02dcd367d16b7ea8469d370861ec86 DOI: 10.1016/j.mito.2012.07.108
2012	Influence of intergeneric/interspecies mitochondrial injection; parthenogenetic development of bovine oocytes after injection of mitochondria derived from somatic cells Takeda K., Srirattana K., Matsukawa K., Akagi S., Kaneda M., Tasai M., Nirasawa K., Pinkert C.A., Parnpai R., Nagai T.	Journal of Reproduction and Development
Abstract: Interspecies/intergeneric mitochondrial heteroplasmy can occur in interspecies/intergeneric hybrid embryos or following nuclear transfer. In the present study, intergeneric buffalo (Bubalus bubalis) mitochondria (WB-mt) or interspecies murine (Mus spretus) mitochondria (M-mt) were injected into bovine (Bos taurus) oocytes, and the subsequent embryonic development was characterized. Fibroblast mitochondria (WB-mt or M-mt) were microinjected into in vitro matured bovine oocytes followed by oocyte activation by a combination of electrical stimulation and 6-dimethylaminopurine treatment. After seven days of culture, embryo development was evaluated. The copy number of specific mtDNA populations (introduced and native mtDNA) from heteroplasmic oocytes was estimated using real-time PCR. The results illustrated that oocytes injected with either WB-mt or M-mt can develop to the blastocyst stage (20.6% and 19.6%). Cleavage division rates and development to the morula stage in oocytes injected with WB-mt were lower (76.2% and 45.9%, respectively) in comparison with uninjected oocytes (89.2% and 59.1%, respectively) (P<0.05). However, no differences were found in comparing M-mt injected oocytes and controls (P>0.05). An increase in bovine mtDNA copy number was observed at the expanded blastocyst stage of injected embryos (P<0.01), while the number of injected mtDNA was stable throughout development. This study demonstrates that interspecies/intergeneric mitochondrial injected bovine oocytes have the ability to develop to the blastocyst stage after parthenogenetic activation and that injected mtDNA was neither selectively destroyed nor enhanced through development. Moreover, injected intergeneric mitochondria had a demonstrated influence on bovine parthenogenetic development and mtDNA replication. © 2012 by the Society for Reproduction and Development. Keyword: Bovine oocyte; Mitochondria; Mouse; mtDNA; Water buffalo Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84863843415&doi=10.1262%2fjrd.2011-013&partnerID=40&md5=614a245278426d3cfb1a9af64b05acc4 DOI: 10.1262/jrd.2011-013
2012	Full-term development of gaur-bovine interspecies somatic cell nuclear transfer embryos: Effect of Trichostatin A treatment Srirattana K., Imsoonthornruksa S., Laowtammathron C., Sangmalee A., Tunwattana W., Thongprapai T., Chaimongkol C., Ketudat-Cairns M., Parnpai R.	Cellular Reprogramming
Abstract: Trichostatin A (TSA) has previously been used in somatic cell nuclear transfer (SCNT) to improve the cloning efficiency in several species, which led our team to investigate the effects of TSA on the full-term development of bovine SCNT and gaur-bovine interspecies SCNT (gaur iSCNT; gaur somatic cells as donors and bovine oocytes as recipients) embryos. Treatment with 50 nM TSA for 10 h after fusion had no positive effects on the rates of fusion, cleavage, or the development to eight-cell or morula stages in both bovine SCNT and gaur iSCNT embryos. However, TSA treatment significantly enhanced the blastocyst formation rate in bovine SCNT embryos (44 vs. 32-34% in the TSA-treated and TSA-untreated groups, respectively), but had no effects on gaur iSCNT embryos. The fresh blastocysts derived from bovine SCNT and gaur iSCNT embryos (fresh groups), as well as vitrified bovine SCNT blastocysts (vitrified group), were transferred to bovine recipients. We found that TSA treatment increased the pregnancy rates only in recipients receiving fresh bovine SCNT embryos. In recipients receiving TSA-treated bovine SCNT embryos, three cloned calves from the fresh group and twin cloned calves from the vitrified group were delivered; however, no calf was born from the TSA-untreated bovine SCNT embryos. In contrast, one gaur iSCNT calf was born from a recipient receiving blastocysts from the TSA-untreated group. In summary, TSA improved the preimplantation development and pregnancy rates of bovine SCNT embryos, but did not have any beneficial effect on gaur iSCNT embryos. However, one gaur iSCNT calf reached full-term development. © Mary Ann Liebert, Inc. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84862181421&doi=10.1089%2fcell.2011.0099&partnerID=40&md5=3757f5285581535bcf68e9e36db3b118 DOI: 10.1089/cell.2011.0099
2012	Development of intergeneric and intrageneric somatic cell nuclear transfer (SCNT) cat embryos and the determination of telomere length in cloned offspring Imsoonthornruksa S., Sangmalee A., Srirattana K., Parnpai R., Ketudat-Cairns M.	Cellular Reprogramming
Abstract: Somatic cell nuclear transfer (SCNT) holds potential as a useful tool for agricultural and biomedical applications. In vitro development of marbled cat intergeneric SCNT reconstructed into domestic cat cytoplast revealed that cloned, marbled cat embryo development was blocked at the morula stage. No pregnancies resulted from the transfer of one-to eight-cell stage embryos into domestic cat surrogate mothers. This suggested that abnormalities occurred in the cloned marbled cat embryos, which may be associated with incomplete reprogramming during early embryo development. Two pregnancies were established in surrogate mothers that received cloned domestic cat embryos, but SCNT offspring developed abnormally. Some specific phenotypes that were observed included incomplete abdominal wall disclosure, improper fetal development. In addition, some of the fetuses were mummified or stillbirths. The two live births died within 5 days. Telomere lengths of cloned kittens as determined by qualtitative polymerase chain reaction (qPCR) were inconclusive: some were found to be shorter, longer, or the same as donor control cells. Our findings support the hypothesis that telomere lengths do not govern the health of these cloned animals. A lack of complete reprogramming may lead to developmental failure and the abnormalities observed in cloned offspring. © 2012, Mary Ann Liebert, Inc. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84856753208&doi=10.1089%2fcell.2011.0054&partnerID=40&md5=8cfc293b046c62b7631a46103fd996b0 DOI: 10.1089/cell.2011.0054
2011	The effects of manipulation medium, culture system and recipient cytoplast on in vitro development of intraspecies and intergeneric felid embryos Imsoonthornruksa S., Lorthongpanich C., Sangmalee A., Srirattana K., Laowtammathron C., Tunwattana W., Somsa W., Ketudat-Cairns M., Nagai T., Parnpai R.	Journal of Reproduction and Development
Abstract: The aim of this study was to investigate if reconstructed felid embryos obtained by intraspecies or intergeneric cloning can develop in vitro. Fibroblast cells (f) from a domestic cat (DCf), marbled cat (MCf) and bovine (Bf) were used as donor cells, and oocytes (o) from domestic cats (DCo) and bovine (Bo) were used as recipient cytoplasts. There were two intraspecies (donor cell + recipient cytoplast: DCf + DCo and Bf + Bo) and three intergeneric (MCf + DCo, DCf + Bo and MCf + Bo) cloning groups in the study. In Experiment 1, the effects of manipulation media, modified TCM-199 (199H) or Emcare holding medium (EHM), on in vitro development of DCf + DCo embryos were investigated. The blastocyst formation rate (BFR) of the embryos manipulated in EHM (33.3%) was higher (P<0.05) compared with those manipulated in 199H (18.1%). In Experiment 2, DCf + DCo and MCf + DCo embryos were cocultured with or without domestic cat oviductal epithelium cells. Irrespective of coculture, the same BFR was obtained for DCf + DCo embryos (44.4 vs. 38.0%), while MCf + DCo embryos could not develop beyond the morula stage. In experiment 3, although the development of MCf + DCo and DCf + Bo embryos was arrested at the morula stage, 8.6% of MCf + Bo embryos were able to develop to the blastocyst stage. These results demonstrated that EHM was superior to 199H as an embryo manipulation medium and that the DCo and Bo could support the early embryonic development of intergeneric cloned marbled cat embryos up to the morula stage. However, postimplantation development still needs to be investigated. © 2011 by the Society for Reproduction and Development. Keyword: Bovine cytoplast; Development; Felid embryos; Intergeneric cloning; Marbled cat Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-79960139959&doi=10.1262%2fjrd.10-108H&partnerID=40&md5=cb4879aabfaddd4c2ef855b8ae23f449 DOI: 10.1262/jrd.10-108H
2011	Spectroscopic signature of mouse embryonic stem cell-derived hepatocytes using synchrotron Fourier transform infrared microspectroscopy Thumanu K., Tanthanuch W., Ye D., Sangmalee A., Lorthongpanich C., Parnpai R., Heraudd P.	Journal of Biomedical Optics
Abstract: Stem cell-based therapy for liver regeneration has been proposed to overcome the persistent shortage in the supply of suitable donor organs. A requirement for this to succeed is to find a rapid method to detect functional hepatocytes, differentiated from embryonic stem cells. We propose Fourier transform infrared (FTIR) microspectroscopy as a versatile method to identify the early and last stages of the differentiation process leading to the formation of hepatocytes. Using synchrotron-FTIR microspectroscopy, the means of identifying hepatocytes at the single-cell level is possible and explored. Principal component analysis and subsequent partial leastsquares (PLS) discriminant analysis is applied to distinguish endoderm induction from hepatic progenitor cells and matured hepatocyte-like cells. The data are well modeled by PLS with endoderm induction, hepatic progenitor cells, and mature hepatocyte-like cells able to be discriminated with very high sensitivity and specificity. This method provides a practical tool to monitor endoderm induction and has the potential to be applied for quality control of cell differentiation leading to hepatocyte formation. © 2011 Society of Photo-Optical Instrumentation Engineers (SPIE). Keyword: Fourier transform infrared microspectroscopy; Hepatocytes; Stem cells Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-80455163221&doi=10.1117%2f1.3580253&partnerID=40&md5=92a9079069ff907e1793f5540a00f388 DOI: 10.1117/1.3580253