Assoc.Prof. Rangsun Parnpai

Year	Title	Journal
2020	Effects of L-carnitine on embryo development of vitrified swamp buffalo oocytes following in vitro fertilization Liang Y., Yoisungnern T., Huang Y., Parnpai R.	Livestock Science
Abstract: The aim of this study was to evaluate the effect of L-carnitine (LC) supplemented in vitro maturation (IVM) medium on the cryotolerance and development of buffalo oocytes. Mitochondrial activity, reactive oxygen species (ROS) content, and lipid droplet distribution of vitrified-warmed buffalo oocytes were also observed. Oocytes were matured in IVM medium supplemented with 0, 0.3, 0.6, and 1.2 mg/mL LC and some of them were vitrified by the Cryotop® method. The highest survival, morula, and blastocyst rates were obtained with 0.6 mg/mL LC, irrespective of whether oocytes were fresh or vitrified-warmed. The density of active mitochondria in vitrified oocytes was significantly higher with 0.6 mg/mL LC than without LC treatment. In contrast, H2O2 levels and lipid droplet content were significantly lower following LC treatment than without it. In conclusion, supplementation with 0.6 mg/mL LC during IVM improves the buffalo oocytes’ survival rate and subsequent embryo development after in vitro fertilization. This may be linked to improved mitochondrial activity, enhanced β-oxidation, and reduced ROS levels. © 2020 Keyword: Buffalo; L-carnitine; oocytes; Vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85078142635&doi=10.1016%2fj.livsci.2020.103933&partnerID=40&md5=92e6688e3ae09b83472a9c182f6341c2 DOI: 10.1016/j.livsci.2020.103933
2020	A Xeno-Free Strategy for Derivation of Human Umbilical Vein Endothelial Cells and Wharton's Jelly Derived Mesenchymal Stromal Cells: A Feasibility Study toward Personal Cell and Vascular Based Therapy Kunkanjanawan H., Kunkanjanawan T., Khemarangsan V., Yodsheewan R., Theerakittayakorn K., Parnpai R.	Stem Cells International
Abstract: Coimplantation of endothelial cells (ECs) and mesenchymal stromal cells (MSCs) into the transplantation site could be a feasible option to achieve a sufficient level of graft-host vascularization. To find a suitable source of tissue that provides a large number of high-quality ECs and MSCs suited for future clinical application, we developed a simplified xeno-free strategy for isolation of human umbilical vein endothelial cells (HUVECs) and Wharton's jelly-derived mesenchymal stromal cells (WJ-MSCs) from the same umbilical cord. We also assessed whether the coculture of HUVECs and WJ-MSCs derived from the same umbilical cord (autogenic cell source) or from different umbilical cords (allogenic cell sources) had an impact on in vitro angiogenic capacity. We found that HUVECs grown in 5 ng/ml epidermal growth factor (EGF) supplemented xeno-free condition showed higher proliferation potential compared to other conditions. HUVECs and WJ-MSCs obtained from this technic show an endothelial lineage (CD31 and von Willebrand factor) and MSC (CD73, CD90, and CD105) immunophenotype characteristic with high purity, respectively. It was also found that only the coculture of HUVEC/WJ-MSC, but not HUVEC or WJ-MSC mono-culture, provides a positive effect on vessel-like structure (VLS) formation, in vitro. Further investigations are needed to clarify the pros and cons of using autogenic or allogenic source of EC/MSC in tissue engineering applications. To the best of our knowledge, this study offers a simple, but reliable, xeno-free strategy to establish ECs and MSCs from the same umbilical cord, a new opportunity to facilitate the development of personal cell-based therapy. © 2020 Hataiwan Kunkanjanawan et al. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85091703702&doi=10.1155%2f2020%2f8832052&partnerID=40&md5=aa4693b85f183a3260393541346822ff DOI: 10.1155/2020/8832052
2020	The effects of vitrification after equilibration in different concentrations of cryoprotectants on the survival and quality of bovine blastocysts Yodrug T., Parnpai R., Hirao Y., Somfai T.	Animal Science Journal
Abstract: This study assessed the effects of cryoprotectant concentration during equilibration on the efficiency of bovine blastocyst vitrification and the expression of selected developmentally important genes. In vitro produced bovine blastocysts were equilibrated in either 7.5% ethylene glycol (EG) + 7.5% DMSO (Va group) or in 2% EG + 2% DMSO (Vb group) then vitrified on Cryotop® sheets in 16.5% EG + 16.5% DMSO + 0.5M sucrose. After warming, embryos were cultured for 48 hr. Re-expansion, hatching, and the numbers of total and membrane damaged cells were compared among vitrified groups and a control. There was no significant difference between the vitrified groups in survival, cell numbers and the extent of membrane damage. Vitrification increased the number of membrane-damaged cells in both groups, however, in a greater extent in the Vb group. Vitrification increased (p <.05) the expression of the HSP70 gene in Va but not in Vb embryos. The expression of IGF2R, SNRPN, HDAC1, DNMT3B, BAX, OCT4, and IFN-t genes were the same in control and vitrified groups. In conclusion, the concentration of cryoprotectants during equilibration did not affect survival rates; however, normal cell numbers could be maintained only by equilibration in 15% cryoprotectants which was associated with increased HSP70 expression. © 2020 Japanese Society of Animal Science Keyword: bovine embryo; equilibration; gene expression; viability; vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85091052934&doi=10.1111%2fasj.13451&partnerID=40&md5=b6ea01fe09ce02761d15610a2f9d09e1 DOI: 10.1111/asj.13451
2019	Enhanced hepatogenic differentiation of human wharton’s jelly-derived mesenchymal stem cells by using three-step protocol Panta W., Imsoonthornruksa S., Yoisungnern T., Suksaweang S., Ketudat-Cairns M., Parnpai R.	International Journal of Molecular Sciences
Abstract: Currently, human Wharton's jelly-derived mesenchymal stem cells (hWJ-MSCs) are an attractive source of stem cells for cell-based therapy, owing to their ability to undergo self-renewal and differentiate into all mesodermal, some neuroectodermal, and endodermal progenies, including hepatocytes. Herein, this study aimed to investigate the effects of sodium butyrate (NaBu), an epigenetic regulator that directly inhibits histone deacetylase, on hepatic endodermal lineage differentiation of hWJ-MSCs. NaBu, at 1 mM, optimally promoted endodermal differentiation of hWJ-MSCs, along with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) supplementation (EGF + bFGF + 1 mM NaBu). CXCR4, HNF3β, SOX17 (endodermal), and GATA6 (mesendodermal) mRNAs were also up-regulated (p < 0.001). Immunocytochemistry and a Western blot analysis of SOX17 and HNF3β confirmed that the EGF + bFGF + 1 mM NaBu condition was appropriately pre-treated with hWJ-MSCs before hepatogenic differentiation. Furthermore, the hepatogenic medium + NaBu pre-treatment up-regulated hepatoblast (AFP and HNF3β) and hepatic (CK18 and ALB) markers, and increased the proportion of mature hepatocyte functions, including G6P, C/EBPα, and CYP2B6 mRNAs, glycogen storage and urea secretion. The hepatogenic medium + NaBu in the pre-treatment step can induce hWJ-MSC differentiation toward endodermal, hepatoblastic, and hepatic lineages. Therefore, the hepatogenic medium + NaBu pre-treatment for differentiating hWJ-MSCs could represent an alternative protocol for cell-based therapy and drug screening in clinical applications. © 2019 by the authors. Licensee MDPI, Basel, Switzerland. Keyword: Endoderm; Hepatocyte-like cells; Histone deacetylase inhibitor; Sodium butyrate; Wharton’s jelly mesenchymal stem cells Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85068566478&doi=10.3390%2fijms20123016&partnerID=40&md5=efddb5bbd54926c595277e43d7e3f269 DOI: 10.3390/ijms20123016
2018	Effect of vitrification procedures on the subsequent development of in vitro matured swamp buffalo oocytes following in vitro fertilization Liang Y.Y., Parnpai R.	Animal Science Journal
Abstract: Nowadays, the efficiency of buffalo oocytes cryopreservation is still low. The purpose of this study was to evaluate effects of two combinations of cryoprotectant agents (CPAs) and two vitrification devices for vitrification of swamp buffalo oocytes on their survival after vitrification warming, and subsequent developmental ability after in vitro fertilization. In vitro matured (IVM) oocytes were vitrified by either Cryotop (CT) or solid surface vitrification (SSV) interacting with vitrification solution A (VA) or B (VB). In the VA or VB solution exposed test, the oocytes showed similar survival rates, but decreased blastocyst rates after in vitro fertilization compared with that of untreated oocytes. After vitrification, the CT method combined with VA solution yielded a higher survival rate (91.3 ± 5.84%) of vitrified oocytes than that combined with VB solution (69.8 ± 4.19%–75.8 ± 4.55%); however, all the vitrification treatments showed lower blastocyst rates (1.1 ± 0.07%–5.2 ± 0.24%) compared with that of untreated oocytes (18.0 ± 1.09%). Our results indicated that combined vitrification treatments in this study did not improve the decreased ability of vitrified oocytes developing to the blastocyst stage. © 2018 Japanese Society of Animal Science Keyword: IVF; oocytes; swamp buffalo; vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85051129341&doi=10.1111%2fasj.13044&partnerID=40&md5=b6b6472bfbafb50b981b649885fc6b9f DOI: 10.1111/asj.13044
2018	New insights on water buffalo genomic diversity and post-domestication migration routes from medium density SNP chip data Colli L., Milanesi M., Vajana E., Iamartino D., Bomba L., Puglisi F., Corvo M.D., Nicolazzi E.L., Ahmed S.S.E., Herrera J.R.V., Cruz L., Zhang S., Liang A., Hua G., Yang L., Hao X., Zuo F., Lai S.-J., Wang S., Liu R., Gong Y., Mokhber M., Mao Y., Guan F., Vlaic A., Vlaic B., Ramunno L., Cosenza G., Ahmad A., Soysal I., Ünal E.Ö., Ketudat-Cairns M., Garcia J.F., Utsunomiya Y.T., Baruselli P.S., Amaral M.E.J., Parnpai R., Drummond M.G., Galbusera P., Burton J., Hoal E., Yusnizar Y., Sumantri C., Moioli B., Valentini A., Stella A., Williams J.L., Ajmone-Marsan P.	Frontiers in Genetics
Abstract: The domestic water buffalo is native to the Asian continent but through historical migrations and recent importations, nowadays has a worldwide distribution. The two types of water buffalo, i.e., river and swamp, display distinct morphological and behavioral traits, different karyotypes and also have different purposes and geographical distributions. River buffaloes from Pakistan, Iran, Turkey, Egypt, Romania, Bulgaria, Italy, Mozambique, Brazil and Colombia, and swamp buffaloes from China, Thailand, Philippines, Indonesia and Brazil were genotyped with a species-specific medium-density 90K SNP panel. We estimated the levels of molecular diversity and described population structure, which revealed historical relationships between populations and migration events. Three distinct gene pools were identified in pure river as well as in pure swamp buffalo populations. Genomic admixture was seen in the Philippines and in Brazil, resulting from importations of animals for breed improvement. Our results were largely consistent with previous archeological, historical and molecular-based evidence for two independent domestication events for river- and swamp-type buffaloes, which occurred in the Indo-Pakistani region and close to the China/Indochina border, respectively. Based on a geographical analysis of the distribution of diversity, our evidence also indicated that the water buffalo spread out of the domestication centers followed two major divergent migration directions: river buffaloes migrated west from the Indian sub-continent while swamp buffaloes migrated from northern Indochina via an east-south-eastern route. These data suggest that the current distribution of water buffalo diversity has been shaped by the combined effects of multiple migration events occurred at different stages of the post-domestication history of the species. © 2018 Colli, Milanesi, Vajana, Iamartino, Bomba, Puglisi, Del Corvo, Nicolazzi, Ahmed, Herrera, Cruz, Zhang. Keyword: Bubalus bubalis; Domestication; Evolutionary history; Genomic diversity; River buffalo; SNP; Swamp buffalo Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85042706895&doi=10.3389%2ffgene.2018.00053&partnerID=40&md5=e8a79d5977c55e648e08824e5f0bb2d5 DOI: 10.3389/fgene.2018.00053
2018	Vitrification of bovine matured oocytes and blastocysts in a paper container Paul A.K., Liang Y., Srirattana K., Nagai T., Parnpai R.	Animal Science Journal
Abstract: In the present study, we aimed to determine the applicability of a paper container for the vitrification of in vitro matured (IVM) bovine oocytes. In experiment 1, IVM oocytes were exposed to vitrification solution (20% dimethylsulfoxide (DMSO), 20% ethylene glycol (EG), and 5 mol/L sucrose), using a two-step method, for 30 s; loaded onto either a paper container or Cryotop; and stored in liquid nitrogen. No significant difference (P < 0.05) in the survival and blastocyst formation rates after in vitro vitrification was observed between the paper container and Cryotop. In experiment 2, IVM oocytes were exposed to either a two- or three-step vitrification solution. The three-step vitrification solution was not significantly different from the two-step solution in terms of oocyte survival, cleavage and blastocyst rates. In experiment 3, in vitro produced blastocysts were graded according to the manual of the International Embryo Transfer Society (grades 1 and 2) and vitrified using the two- and three-step methods. For grade 2 blastocysts, the three-step method showed significantly higher (P < 0.05) survival and hatched blastocyst rates than the two-step method, whereas for grade 1 blastocysts, no significant difference was observed. In conclusion, the paper device and three-step technique are suitable for oocytes and embryo vitrification. © 2017 Japanese Society of Animal Science Keyword: blastocysts; oocytes; paper device; vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85031100168&doi=10.1111%2fasj.12892&partnerID=40&md5=b8b15cb5ec81e8d73b170578844d4250 DOI: 10.1111/asj.12892
2018	Survival and developmental competence of bovine embryos at different developmental stages and separated blastomeres after vitrification in different solutions Juanpanich T., Suttirojpattana T., Takayama M., Liang Y., Dochi O., Parnpai R., Imai K.	Animal Science Journal
Abstract: Generating techniques to enhance the success of blastomere separation is important for bovine economy, because it increases the number of transferable embryos. This study aimed to identify the optimum cryoprotectants for the vitrification of bovine embryos and the separation of blastomeres at different stages. In experiment 1, expanded blastocysts were vitrified in two different vitrification solutions, either (1) ethylene glycol (EG) + propylene glycol (PG) or (2) EG. The survival rate of blastocysts in the EG + PG was higher than that of the EG. In experiment 2, intact two-cell and eight-cell stage embryos were vitrified in the same solutions used in experiment 1. The EG + PG produced more dead embryos than the EG (P < 0.05). In the EG, the rate of blastocyst formation was similar for the vitrified two- and eight-cell embryos and the non-vitrified ywo-cell embryos. In experiment 3, separated blastomeres of two- and eight-cell embryos were vitrified in EG. There was no difference in the rate of blastocyst formation and total number of cells between the two vitrified groups. In summary, at the blastocyst stage, EG + PG was superior, based on both survival rates and cell numbers; however, at the 2–8 cell stage, the use of EG alone was better than the other groups. © 2017 Japanese Society of Animal Science. Keyword: bovine; early stages; embryo development; separated blastomere; vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85040721309&doi=10.1111%2fasj.12890&partnerID=40&md5=2e8c7cb3540fa572970bd37b3fb77c79 DOI: 10.1111/asj.12890
2018	Effects of gel-embedded embryos on developmental competence of separated bovine blastomeres Juanpanich T., Suttirojpattana T., Liang Y., Dochi O., Parnpai R., Imai K.	Livestock Science
Abstract: This study aimed to examine how gel embedding compared with the Well-of-Well (WOW) system when culturing bovine separated blastomeres, in terms of developmental competence rates and blastocyst quality. We first optimized the gel-embedding method via culturing intact zygotes in either 1% agarose or 1% calcium alginate gel. Gel-embedded groups and control did not differ in development rates, but the 1% calcium alginate group was selected for subsequent experiments due to higher blastocyst recovery. The separated embryo group had higher potential for blastocyst production than the intact embryo group. Among separated blastomeres (2- and 8-cell), WOW and 1% calcium alginate did not differ in blastocyst formation rate, except between the 2-cell alginate and WOW groups, with the latter exhibiting the highest rate of blastocyst formation. Within the WOW system, the separated 2-cell embryo resulted in significantly higher rates of OPU (ovum pick-up)-derived blastocysts than the intact 2-cell embryo; however, the latter had more total cells than the former. In conclusion, the 1% calcium alginate gel can support cell growth of separated 2- and 8-cell bovine embryos during blastomere aggregation. Moreover, the gel-embedded technique is a suitable replacement for WOW in producing blastocysts from bovine separated blastomeres. © 2017 Keyword: Agar embedded embryo; Blastomere separation; Bovine; Embryo development Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85034046375&doi=10.1016%2fj.livsci.2017.11.010&partnerID=40&md5=376c8109fd26718939988dbc61689f0b DOI: 10.1016/j.livsci.2017.11.010
2017	Effect of storage tube material and resveratrol during liquid storage of matured bovine oocytes on subsequent development Suttirojpattana T., Somfai T., Matoba S., Nagai T., Parnpai R., Geshi M.	Acta Veterinaria Hungarica
Abstract: This study determined the optimum storage vessel and the effects of resveratrol for the storage of in vitro matured (IVM) bovine oocytes. After IVM, the oocytes were kept in a Hepes-buffered medium at 25 °C for 20 h in different containers including Eppendorf tubes (ET) made of polypropylene (PP) and polystyrene (PS), and tissue culture tubes (TCT) made of PP, PS, and glass. Then oocytes were subjected to IVF and subsequent in vitro embryo development was compared among the groups and to that of a control group without storage. The percentage of blastocyst development in the control group was significantly higher than in the stored groups (P < 0.05). Among oocytes stored in TCT, the percentage of blastocyst development of oocytes stored in glass TCT was significantly higher than that of oocytes stored in PP and PS TCT (P < 0.05); however, it did not differ from that of oocytes stored in ET. The quality of blastocysts did not differ among the control and stored groups. Embryo development was not affected when 0.1, 1 or 10 μM resveratrol was added to the medium during oocyte storage. In conclusion, glass tubes were optimal for oocyte storage and resveratrol did not improve the development of stored oocytes. © 2017 Akadémiai Kiadó, Budapest. Keyword: Bovine; Containers; Oocyte; Resveratrol; Storage Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85038638470&doi=10.1556%2f004.2017.053&partnerID=40&md5=3fb0dd28471140150836b53c1c74aab2 DOI: 10.1556/004.2017.053