Assoc.Prof. Rangsun Parnpai

Year	Title	Journal
2016	Effect of Chromatin-Remodeling Agents in Hepatic Differentiation of Rat Bone Marrow-Derived Mesenchymal Stem Cells in Vitro and in Vivo Ye D., Li T., Heraud P., Parnpai R.	Stem Cells International
Abstract: Epigenetic events, including covalent histone modifications and DNA methylation, play fundamental roles in the determination of lineage-specific gene expression and cell fates. The aim of this study was to determine whether the DNA methyltransferase inhibitor (DNMTi) 5-aza-2′-deoxycytidine (5-aza-dC) and the histone deacetylase inhibitor (HDACi) trichostatin A (TSA) promote the hepatic differentiation of rat bone marrow-derived mesenchymal stem cells (rBM-MSCs) and their therapeutic effect on liver damage. 1 μM TSA and 20 μM 5-aza-dC were added to standard hepatogenic medium especially at differentiation and maturation steps and their potential function on hepatic differentiation in vitro and in vivo was determined. Exposure of rBM-MSCs to 1 μM TSA at both the differentiation and maturation steps considerably improved hepatic differentiation. TSA enhanced the development of the hepatocyte shape, promoted the chronological expression of hepatocyte-specific markers, and improved hepatic functions. In contrast, treatment of rBM-MSCs with 20 μM 5-aza-dC alone or in combination with TSA was ineffective in improving hepatic differentiation in vitro. TSA and/or 5-aza-dC derived hepatocytes-like cells failed to improve the therapeutic potential in liver damage. We conclude that HDACis enhance hepatic differentiation in a time-dependent manner, while DNMTis do not induce the hepatic differentiation of rBM-MSCs in vitro. Their in vivo function needs further investigation. © 2016 Danna Ye et al. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84971633740&doi=10.1155%2f2016%2f3038764&partnerID=40&md5=a0885a5f6c58d4b2d7b268ed6d7901e3 DOI: 10.1155/2016/3038764
2016	Induction of mESCs into Hepatic Stem Cells by using Embryonic Chicken Hearts Suksaweang S., Ye D., Parnpai R.	Journal of the Medical Association of Thailand = Chotmaihet thangphaet
Abstract: Background: Many researchers have been trying different methods for obtaining stem cells. Some studies have failed due to the growth of a tumor after stem cells transplantation. Several successful tries for getting stem cells or stem cell like cells: direct isolation from tissue, direct isolation from blood or fluids, iPS cells, small molecules induced stem cells. However, none have used real organ stimulation in the induction of a specific stem cell lineage.Objective: To induce a lineage specific hepatic stem cell using isolated embryonic organs.Material and Method: The embryonic stem cells were cultured through confluence. After observing several colonies formations, we put freshly isolated chicken embryonic hearts onto the colonies. After, at least, four days, we started looking for hepatic plate-like formations.Results: After several trials, we found that the chicken embryonic hearts, on day 4, could actually induce a hepatic cell fate for the mouse embryonic stem cells. We were able to show specific marker for early hepatic lineage such as the production of Albumin, AFP. When these cells were tested for a hepatocyte function, we found glycogen formation inside the cells.Conclusion: Isolated early embryonic chicken hearts are acceptable for inducing embryonic stem cells into the hepatic stem cell lineage. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85049294491&partnerID=40&md5=cdb7912213f66bc3a8c9a7b546ca3353 DOI:
2016	Comparison of methods for deriving neural progenitor cells from nonhuman primate embryonic stem cells Khlongkhlaeo A., Carter R.L., Parnpai R., Chan A.W.S.	Asian Biomedicine
Abstract: Background: Feeder-free monolayer culture and the suspension culture of embryoid bodies (EB) followed by adherent culture, rosette selection, and expansion are 2 methods for deriving neural progenitor cells (NPCs). Direct comparison of these 2 methods has not yet been reported. Objectives: To compare the influence of NPC derivation methods on the properties of NPCs derived from rhesus monkey (Macaca mulatta) embryonic stem cells (rhESCs). Methods: rhESCs were used to derive NPC lines using 2 different methods. EB were produced from a suspension culture of rhESC clumps and rhESCs were cultured in feeder-free monolayers on poly-L-ornithine/laminin coated plates. NPCs were derived by exposure to induction factors. Cell morphology, neural and nonneural lineage markers were evaluated. We measured the expression of nuclear receptor tailless (TLX), which acts as repressor of glial fibrillary acidic protein (GFAP) expression. Results: NPCs were successful derived using either method, with homogenous populations based on the expression of nestin (>97%) and Pax6 (>99%) as shown by flow cytometry. No significant difference in NPC specific markers or ability to differentiate into neurons in vitro was found between the methods. However, the expression of GFAP was >400-fold higher in cells produced by the feeder-free method. This distinction was consistent with the lower expression of TLX. Conclusions: NPCs derived by feeder-free and EB methods share similar morphology and properties. The elevated expression of GFAP and reduced expression of TLX in NPCs derived using the feeder-free method may explain their greater heterogeneity and tendency to differentiate toward cells of an astrocyte lineage. Keyword: Embryonic stem cells; Neural progenitor cell derivation; Neural progenitor cells; Nonhuman primate; TLX Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85007240219&doi=10.5372%2f1905-7415.1005.505&partnerID=40&md5=b826cab776f908bf604031c48775c089 DOI: 10.5372/1905-7415.1005.505
2015	Expression and Purification of Recombinant Human Basic Fibroblast Growth Factor Fusion Proteins and Their Uses in Human Stem Cell Culture Imsoonthornruksa S., Pruksananonda K., Parnpai R., Rungsiwiwut R., Ketudat-Cairns M.	Journal of Molecular Microbiology and Biotechnology
Abstract: To reduce the cost of cytokines and growth factors in stem cell research, a simple method for the production of soluble and biological active human basic fibroblast growth factor (hbFGF) fusion protein in Escherichia coli was established. Under optimal conditions, approximately 60-80 mg of >95% pure hbFGF fusion proteins (Trx-6xHis-hbFGF and 6xHis-hbFGF) were obtained from 1 liter of culture broth. The purified hbFGF proteins, both with and without the fusion tags, were biologically active, which was confirmed by their ability to stimulate proliferation of NIH3T3 cells. The fusion proteins also have the ability to support several culture passages of undifferentiated human embryonic stem cells and induce pluripotent stem cells. This paper describes a low-cost and uncomplicated method for the production and purification of biologically active hbFGF fusion proteins. © 2016 S. Karger AG, Basel. Keyword: Fusion protein; Human basic fibroblast growth factor; Human embryonic stem cells; Induced pluripotent stem cells; Recombinant protein Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84947811001&doi=10.1159%2f000441453&partnerID=40&md5=3afb4b44d703c38e4f63fa571f0f575c DOI: 10.1159/000441453
2015	Comparison of Cryotop and micro volume air cooling methods for cryopreservation of bovine matured oocytes and blastocysts Punyawai K., Anakkul N., Srirattana K., Aikawa Y., Sangsritavong S., Nagai T., Imai K., Parnpai R.	Journal of Reproduction and Development
Abstract: This study was designed to compare the efficiency of the Cryotop method and that of two methods that employ a micro volume air cooling (MVAC) device by analyzing the survival and development of bovine oocytes and blastocysts vitrified using each method. In experiment I, in vitro-matured (IVM) oocytes were vitrified using an MVAC device without direct contact with liquid nitrogen (LN2; MVAC group) or directly plunged into LN2 (MVAC in LN2 group). A third group of IVM oocytes was vitrified using a Cryotop device (Cryotop group). After warming, vitrified oocytes were fertilized in vitro. There were no significant differences in cleavage and blastocyst formation rates among the three vitrified groups, with the rates ranging from 53.1% to 56.6% and 20.0% to 25.5%, respectively; however, the rates were significantly lower (P < 0.05) than those of the fresh control group (89.3% and 43.3%, respectively) and the solution control group (87.3% and 42.0%, respectively). In experiment II, in vitro-produced (IVP) expanded blastocysts were vitrified using the MVAC, MVAC in LN2 and Cryotop methods, warmed and cultured for survival analysis and then compared with the solution control group. The rate of development of vitrified-warmed expanded blastocysts to the hatched blastocyst stage after 24 h of culture was lower in the MVAC in LN2 group than in the solution control group; however, after 48–72 h of culture, the rates did not significantly differ between the groups. These results indicate that the MVAC method without direct LN2 contact is as effective as the standard Cryotop method for vitrification of bovine IVM oocytes and IVP expanded blastocysts. © 2015 by the Society for Reproduction and Development. Keyword: Blastocysts; Cryodevice; Cryotop; Micro volume air cooling; Oocytes Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84944554714&doi=10.1262%2fjrd.2014-163&partnerID=40&md5=3c0fe5d8f695a2358b81dc2c4ba86eb7 DOI: 10.1262/jrd.2014-163
2015	Pretreatment of in vitro matured bovine oocytes with docetaxel before vitrification: Effects on cytoskeleton integrity and developmental ability after warming Chasombat J., Nagai T., Parnpai R., Vongpralub T.	Cryobiology
Abstract: The stabilization of spindle fibersis important for successful vitrification of bovine oocytes because microtubules and other cytoskeleton fibers (CSF) can be damaged during vitrification, resulting in failure of fertilization after thawing. Docetaxel, a stabilizing agent, could potentially reduce CSF damage of bovine oocytes induced during vitrification. However, there have been no reports on the effects of docetaxel on their vitrification. Experiment 1 was conducted to investigate the effects of various doses of docetaxel (0.0, 0.05, 0.5, 5.0 and 50. μM) in preincubation medium of in vitro matured (IVM) bovine oocytes on their developmental ability after in vitro fertilization (IVF). The results show that 0.05. μM docetaxel had no adverse effect on embryo development, while docetaxel at a concentration of ≥0.5. μM inhibited development. Experiments 2 and 3 were conducted to investigate the effects of preincubation of IVM bovine oocytes with 0.05. μM docetaxel for 30. min prior to vitrification-warming on CSF integrity (Experiment 2), and on oocyte survival and viability after IVF (Experiment 3). When preincubated with 0.05. μM docetaxel for 30. min before vitrification, post-thawed oocytes had less CSF damage and higher survival rates compared with those untreated with docetaxel before vitrification. Surviving oocytes also had higher rates of cleavage and development to the blastocyst stage after IVF. In conclusion, preincubation of IVM bovine oocytes with 0.05. μM docetaxel for 30. min before vitrification was effective at preventing CSF damage during vitrification, and improving oocyte viability after warming and subsequent cleavage and blastocyst formation after IVF. © 2015 Elsevier Inc. Keyword: Bovine oocytes; Cytoskeleton fibers; Docetaxel; Vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84942296757&doi=10.1016%2fj.cryobiol.2015.07.002&partnerID=40&md5=2558f2c6bb9470267e433f5cd8974aba DOI: 10.1016/j.cryobiol.2015.07.002
2015	Internalization of silver nanoparticles into mouse spermatozoa results in poor fertilization and compromised embryo development Yoisungnern T., Choi Y.-J., Woong Han J., Kang M.-H., Das J., Gurunathan S., Kwon D.-N., Cho S.-G., Park C., Kyung Chang W., Chang B.-S., Parnpai R., Kim J.-H.	Scientific Reports
Abstract: Silver nanoparticles (AgNPs) have many features that make them attractive as medical devices, especially in therapeutic agents and drug delivery systems. Here we have introduced AgNPs into mouse spermatozoa and then determined the cytotoxic effects of AgNPs on sperm function and subsequent embryo development. Scanning electron microscopy and transmission electron microscopy analyses showed that AgNPs could be internalized into sperm cells. Furthermore, exposure to AgNPs inhibited sperm viability and the acrosome reaction in a dose-dependent manner, whereas sperm mitochondrial copy numbers, morphological abnormalities, and mortality due to reactive oxygen species were significantly increased. Likewise, sperm abnormalities due to AgNPs internalization significantly decreased the rate of oocyte fertilization and blastocyst formation. Blastocysts obtained from AgNPs-treated spermatozoa showed lower expression of trophectoderm-associated and pluripotent marker genes. Overall, we propose that AgNPs internalization into spermatozoa may alter sperm physiology, leading to poor fertilization and embryonic development. Such AgNPs-induced reprotoxicity may be a valuable tool as models for testing the safety and applicability of medical devices using AgNPs. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84930959425&doi=10.1038%2fsrep11170&partnerID=40&md5=f7745d382812746d322eed94494d473a DOI: 10.1038/srep11170
2015	Bovine embryo sex determination by multiplex loop-mediated isothermal amplification Khamlor T., Pongpiachan P., Parnpai R., Punyawai K., Sangsritavong S., Chokesajjawatee N.	Theriogenology
Abstract: In cattle, the ability to determine the sex of embryos before embryo transfer is beneficial for increasing the number of animals with the desired sex. This study therefore developed a new modification of loop-mediated isothermal amplification in a multiplex format (multiplex LAMP) for highly efficient bovine embryo sexing. Two chromosomal regions, one specific for males (Y chromosome, S4 region) and the other common to both males and females (1.715 satellite DNA), were amplified in the same reaction tube. Each target was amplified by specifically designed inner primers, outer primers, and loop primers, where one of the S4 loop primers was labeled with the fluorescent dye 6-carboxyl-X-rhodamine (emitting a red color), whereas both satellite loop primers were labeled with the fluorescent dye fluorescein isothiocyanate (emitting a green color). After amplification at 63°C for 1hour, the amplified products were precipitated by a small volume of cationic polymer predispensed inside the reaction tube cap. Green precipitate indicated the presence of only control DNA without the Y chromosome, whereas orange precipitate indicated the presence of both target DNAs, enabling interpretation as female and male, respectively. Accuracy of the multiplex LAMP assay was evaluated using 46 bovine embryos with known sex (25 male and 21 female) generated by somatic cell nuclear transfer and confirmed by multiplex polymerase chain reaction. The multiplex LAMP showed 100% accuracy in identifying the actual sex of the embryos and provides a fast, simple, and cost-effective tool for bovine embryo sexing. © 2015 Elsevier Inc. Keyword: Bovine; Embryo sexing; Multiplex LAMP; Sex determination Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84925082544&doi=10.1016%2fj.theriogenology.2014.11.025&partnerID=40&md5=f265ef846f0e723a007f62270d669313 DOI: 10.1016/j.theriogenology.2014.11.025
2014	Reversal of cellular phenotypes in neural cells derived from Huntington's disease monkey-induced pluripotent stem cells Carter R.L., Chen Y., Kunkanjanawan T., Xu Y., Moran S.P., Putkhao K., Yang J., Huang A.H.C., Parnpai R., Chan A.W.S.	Stem Cell Reports
Abstract: Huntington's disease (HD) is a dominant neurodegenerative disorder caused by the expansion of glutamine residues in the N-terminal region of the huntingtin (HTT) protein. The disease results in progressive neuronal loss, leading to motor, cognitive, and psychiatric impairment. Here, we report the establishment of neural progenitor cell (NPC) lines derived from induced pluripotent stem cells (iPSCs) of transgenic HD monkeys. Upon differentiation to neurons, HD neural cells develop cellular features of HD, including the formation of nuclear inclusions and oligomeric mutant HTT (mHTT) aggregates, as well as increased apoptosis. These phenotypes are rescued by genetic suppression of HTT and pharmacological treatment, demonstrating the ability of our HD cell model to respond to therapeutic treatment. The development and reversal of HD-associated phenotypes in neural cells from HD monkeys provides a unique nonhuman primate (NHP) model for exploring HD pathogenesis and evaluating therapeutics that could be assessed further in HD monkeys. © 2014 The Authors. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84908053201&doi=10.1016%2fj.stemcr.2014.07.011&partnerID=40&md5=3ab523bd6c79d634b25e8726ee0e7e57 DOI: 10.1016/j.stemcr.2014.07.011
2014	In vitro development potentiality of expanded bovine blastocysts subsequent to cryotop vitrification Paul A.K., Liang Y., Nagai T., Parnpai R.	Thai Journal of Veterinary Medicine
Abstract: Embryo cryopreservation is a promising area of study in the field of reproductive research. One of the issues that have recently arisen is the discovery that the growing ability of embryos after cryopreservation varies depending on the culture period of the embryo prior to its cryopreservation. Therefore, the present study was designed to explore the in vitro development potentiality of bovine embryos at day 7 and day 8, as well as their survivability and hatchability after Cryotop vitrification. The blastocyst rate at day 8 (28.1%) was higher than that of day 7 (19.1%). Grade 1 (G1) and grade 2 (G2) expanded blastocysts at day 7 and day 8 were vitrified by Cryotop device using 20% (v/v) DMSO, 20% (v/v) EG and 0.5M. Except for the G2 expanded blastocysts at day 8, we found that the survival rates of vitrified G1 and G2 expanded blastocysts were not significantly different from the control group in both day 7 and day 8. However, the day 7 vitrified embryos showed superior rates of hatchability than those of day 8. In a curve estimation of correlation regressions, the hatching rate of day 7 G1 expanded blastocysts at 48 h showed a strong correlation (R2=0.914) with their survival rate. Therefore, we concluded that the day 7 culture period is the most suitable for vitrification of IVF derived blastocysts. Keyword: Blastocysts; Bovine; In vitro fertilization; Vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84929320982&partnerID=40&md5=ae5cfab7c599e7737ef553699c5acfa0 DOI: