Assoc.Prof. Rangsun Parnpai

Year	Title	Journal
2023	The Efficiency of Neurospheres Derived from Human Wharton’s Jelly Mesenchymal Stem Cells for Spinal Cord Injury Regeneration in Rats Somredngan S., Theerakittayakorn K., Nguyen H.T., Ngernsoungnern A., Ngernsoungnern P., Sritangos P., Ketudat-Cairns M., Imsoonthornruksa S., Keeratibharat N., Wongsan R., Rungsiwiwut R., Parnpai R.	International Journal of Molecular Sciences
Abstract: Spinal cord injury (SCI) causes inflammation and neuronal degeneration, resulting in functional movement loss. Since the availability of SCI treatments is still limited, stem cell therapy is an alternative clinical treatment for SCI and neurodegenerative disorders. Human umbilical cord Wharton’s jelly-derived mesenchymal stem cells (hWJ-MSCs) are an excellent option for cell therapy. This study aimed to induce hWJ-MSCs into neural stem/progenitor cells in sphere formation (neurospheres) by using neurogenesis-enhancing small molecules (P7C3 and Isx9) and transplant to recover an SCI in a rat model. Inducted neurospheres were characterized by immunocytochemistry (ICC) and gene expression analysis. The best condition group was selected for transplantation. The results showed that the neurospheres induced by 10 µM Isx9 for 7 days produced neural stem/progenitor cell markers such as Nestin and β-tubulin 3 through the Wnt3A signaling pathway regulation markers (β-catenin and NeuroD1 gene expression). The neurospheres from the 7-day Isx9 group were selected to be transplanted into 9-day-old SCI rats. Eight weeks after transplantation, rats transplanted with the neurospheres could move normally, as shown by behavioral tests. MSCs and neurosphere cells were detected in the injured spinal cord tissue and produced neurotransmitter activity. Neurosphere-transplanted rats showed the lowest cavity size of the SCI tissue resulting from the injury recovery mechanism. In conclusion, hWJ-MSCs could differentiate into neurospheres using 10 µM Isx9 media through the Wnt3A signaling pathway. The locomotion and tissue recovery of the SCI rats with neurosphere transplantation were better than those without transplantation. © 2023 by the authors. Keyword: human umbilical cord Wharton’s jelly-derived mesenchymal stem cells; neurospheres; recovery; spinal cord injury; transplantation Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85149053611&doi=10.3390%2fijms24043846&partnerID=40&md5=050729ba09d3e126f3c4cd38ffe9da7f DOI: 10.3390/ijms24043846
2022	Signaling Pathways Impact on Induction of Corneal Epithelial-like Cells Derived from Human Wharton’s Jelly Mesenchymal Stem Cells Nguyen H.T., Theerakittayakorn K., Somredngan S., Ngernsoungnern A., Ngernsoungnern P., Sritangos P., Ketudat-Cairns M., Imsoonthornruksa S., Assawachananont J., Keeratibharat N., Wongsan R., Rungsiwiwut R., Laowtammathron C., Bui N.X., Parnpai R.	International Journal of Molecular Sciences
Abstract: Corneal epithelium, the outmost layer of the cornea, comprises corneal epithelial cells (CECs) that are continuously renewed by limbal epithelial stem cells (LESCs). Loss or dysfunction of LESCs causes limbal stem cell deficiency (LSCD) which results in corneal epithelial integrity loss and visual impairment. To regenerate the ocular surface, transplantation of stem cell-derived CECs is necessary. Human Wharton’s jelly derived mesenchymal stem cells (WJ-MSCs) are a good candidate for cellular therapies in allogeneic transplantation. This study aimed to test the effects of treatments on three signaling pathways involved in CEC differentiation as well as examine the optimal protocol for inducing corneal epithelial differentiation of human WJ-MSCs. All-trans retinoic acid (RA, 5 or 10 µM) inhibited the Wnt signaling pathway via suppressing the translocation of β-catenin from the cytoplasm into the nucleus. SB505124 downregulated the TGF-β signaling pathway via reducing phosphorylation of Smad2. BMP4 did not increase phosphorylation of Smad1/5/8 that is involved in BMP signaling. The combination of RA, SB505124, BMP4, and EGF for the first 3 days of differentiation followed by supplementing hormonal epidermal medium for an additional 6 days could generate corneal epithelial-like cells that expressed a CEC specific marker CK12. This study reveals that WJ-MSCs have the potential to transdifferentiate into CECs which would be beneficial for further applications in LSCD treatment therapy. © 2022 by the authors. Licensee MDPI, Basel, Switzerland. Keyword: Corneal epithelial cells; Differentiation; Human; Signaling pathways; Wharton’s jelly mesenchymal stem cells Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85126090509&doi=10.3390%2fijms23063078&partnerID=40&md5=53567b9a14517394706db6cbdcba606d DOI: 10.3390/ijms23063078
2022	Strategies to Improve the Efficiency of Somatic Cell Nuclear Transfer Srirattana K., Kaneda M., Parnpai R.	International Journal of Molecular Sciences
Abstract: Mammalian oocytes can reprogram differentiated somatic cells into a totipotent state through somatic cell nuclear transfer (SCNT), which is known as cloning. Although many mammalian species have been successfully cloned, the majority of cloned embryos failed to develop to term, resulting in the overall cloning efficiency being still low. There are many factors contributing to the cloning success. Aberrant epigenetic reprogramming is a major cause for the developmental failure of cloned embryos and abnormalities in the cloned offspring. Numerous research groups attempted multiple strategies to technically improve each step of the SCNT procedure and rescue abnormal epigenetic reprogramming by modulating DNA methylation and histone modifications, overexpression or repression of embryonic-related genes, etc. Here, we review the recent approaches for technical SCNT improvement and ameliorating epigenetic modifications in donor cells, oocytes, and cloned embryos in order to enhance cloning efficiency. © 2022 by the authors. Licensee MDPI, Basel, Switzerland. Keyword: Cloning efficiency; Embryo; Epigenetic modification; Nuclear reprogramming; Somatic cell nuclear transfer Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85124257106&doi=10.3390%2fijms23041969&partnerID=40&md5=42e26125bed9a72f670dbc74a9481b0d DOI: 10.3390/ijms23041969
2022	Current status of assisted reproductive technologies in buffaloes Srirattana K., Hufana-Duran D., Atabay E.P., Duran P.G., Atabay E.C., Lu K., Liang Y., Chaikhun-Marcou T., Theerakittayakorn K., Parnpai R.	Animal Science Journal
Abstract: Buffaloes are raised by small farm holders primarily as source of draft power owing to its resistance to hot climate, disease, and stress conditions. Over the years, transformation of these animals from draft to dairy was deliberately carried out through genetic improvement program leading to the development of buffalo-based enterprises. Buffalo production is now getting more attention and interest from buffalo raisers due to its socioeconomic impact as well as its contribution to propelling the livestock industry in many developing countries. Reproduction of buffaloes, however, is confronted with huge challenge and concern as being generally less efficient to reproduce compared with cattle due to both intrinsic and extrinsic factors such as poor estrus manifestation, silent heat, marked seasonal infertility, postpartum anestrus, long calving interval, delayed puberty, inherently low number of primordial follicles in their ovaries, high incidence of atresia, and apoptosis. Assisted reproductive technologies (ARTs) are major interventions for the efficient utilization of follicle reserve in buffaloes. The present review focuses on estrus and ovulation synchronization for fixed time artificial insemination, in vitro embryo production, intracytoplasmic sperm injection, cryopreservation of oocytes and embryos, somatic cell nuclear transfer, the factors affecting utilization in various ARTs, and future perspectives in buffaloes. © 2022 The Authors. Animal Science Journal published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Animal Science. Keyword: buffalo; embryo and oocytes cryopreservation; estrus synchronization; in vitro embryo production; SCNT Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85138130298&doi=10.1111%2fasj.13767&partnerID=40&md5=138289fa72f025922f505481a0242f60 DOI: 10.1111/asj.13767
2022	The relationship between reactive oxygen species, DNA fragmentation, and sperm parameters in human sperm using simplified sucrose vitrification with or without triple antioxidant supplementation Juanpanich T., Suttirojpattana T., Parnpai R., Vutyavanich T.	Clinical and Experimental Reproductive Medicine
Abstract: Objective: This study examined whether the addition of triple antioxidants (3A)—10 μM acetyl-L-carnitine, 10 μM N-acetyl-L-cysteine, and 5 μM α-lipoic acid—in freezing-thawing medium during human sperm cryopreservation using the sucrose vitrification (SuV) and liquid nitrogen vapor (Vapor) techniques could improve post-thaw survival of spermatozoa. Methods: We analyzed 30 samples from healthy human sperm donors. Each sample was allocated into one of five groups: fresh control, SuV, SuV+3A, Vapor, and Vapor+3A. The sperm motility, morphology, viability, intracellular and extracellular reactive oxygen species (ROS) levels, and sperm DNA fragmentation (SDF) were evaluated. Results: The cryopreserved spermatozoa had significantly reduced percentages of motility (p<0.05) and viability (p<0.05). Antioxidant supplementation non-significantly improved these parameters (p>0.05). No significant differences were found in sperm morphology between the fresh and frozen-thawed groups (p>0.05). After freezing, the extracellular ROS levels in the frozen-thawed groups were significantly higher (p<0.05) than in the fresh group. However, we did not find any differences in intracellular ROS parameters among these groups (p>0.05). The SDF was higher in the SuV and Vapor groups than in the fresh group, but without statistical significance (p=0.075 and p=0.077, respectively). Conclusion: Cryopreservation had detrimental effects on sperm motility, viability, and extracellular ROS levels, without changing the morphology or intracellular ROS levels. Antioxidant supplementation was slightly effective in preventing SDF in frozen-thawed spermatozoa © 2022. THE KOREAN SOCIETY FOR REPRODUCTIVE MEDICINE Keyword: Antioxidants; Dna fragmentation; Human; Spermatozoa; Sucrose; Vitrification Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85131455701&doi=10.5653%2fcerm.2021.05120&partnerID=40&md5=c33fc1834a42e7548aa51fd819612b6f DOI: 10.5653/cerm.2021.05120
2021	Effect of vitrification at different meiotic stages on epigenetic characteristics of bovine oocytes and subsequently developing embryos Yodrug T., Parnpai R., Hirao Y., Somfai T.	Animal Science Journal
Abstract: Vitrification by the Cryotop method is frequently used for bovine oocyte cryopreservation. Nevertheless, vitrified oocytes still have reduced developmental competency compared with fresh counterparts. The objective of this study was to compare the effect of vitrification either at the germinal vesicle (GV) stage or at the metaphase II (MII) stage on epigenetic characteristics of bovine oocytes and subsequently developing embryos. Our results demonstrated that vitrification of oocytes at each meiotic stage significantly reduced blastocyst development after in vitro fertilization (IVF). However, vitrification at the GV stage resulted in higher blastocyst development than did vitrification at the MII stage. Irrespective of the meiotic stage, oocyte vitrification did not affect 5-methylcytosine (5mC) immunostaining intensity in oocyte DNA. However, at both stages, it caused a similar reduction of 5mC levels in DNA of subsequently developing blastocysts. Oocyte vitrification had no effect on the intensity of H3K9me3 and acH3K9 immunostaining in oocytes and subsequent blastocysts. The results suggest that irrespective of meiotic stage, oocyte vitrification alters global methylation in resultant embryos although such alteration in the oocytes was not detected. Oocyte vitrification might not influence histone acetylation and methylation in oocytes and resultant embryos. Vitrification at the immature stage was more advantageous for blastocyst development than at the mature stage. © 2021 Japanese Society of Animal Science Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85112295316&doi=10.1111%2fasj.13596&partnerID=40&md5=658eb2538296eedb8189b5f6b169ba8f DOI: 10.1111/asj.13596
2021	Vitrification of mouse two-cell and blastocyst stage embryos in simplified closed system using either a hemi-straw or a hollow fiber device Suttirojpattana T., Juanpanich T., Parnpai R., Vutyavanich T.	Animal Science Journal
Abstract: Two-cell stage and blastocyst stage mouse embryos were equilibrated in a medium containing 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO) for 8–15 min. Vitrification was performed in a medium containing 0.5 M sucrose and either 15% EG + 15% DMSO, 17.5% EG + 17.5% DMSO, or 20% EG + 20% DMSO for 30 s. They were then placed either on a hemi-straw (HS) or a hollow fiber vitrification (HFV) device and vitrified by cooled air inside a 0.5-ml straw. In two-cell embryos, a 100% survival rate was obtained from all groups except the 20% HS group (P >.05). All vitrified two-cell groups showed similar rates of blastocyst development to that of fresh control group (P >.05), except 17.5% and 20% HFV groups, which were significantly lower than the other groups (P <.05). In the blastocyst embryos, the HFV groups were divided into two subgroups (non-collapsed; HFV-NC and collapsed; HFV-C blastocyst). Re-expansion rate in 15% HFV-NC, 17.5% HFV-NC, and 15% HFV-C groups was reduced (P <.05), whereas the rest were similar to control. In conclusion, we established a simplified, reliable, and closed system for HFV vitrification applying hemi-straw, which does not require skilled practitioners. © 2021 Japanese Society of Animal Science Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85110964740&doi=10.1111%2fasj.13585&partnerID=40&md5=af695cd3da30d53ced4681e15d9b28e8 DOI: 10.1111/asj.13585
2021	Blastocyst development after fertilization with in vitro spermatids derived from nonhuman primate embryonic stem cells Khampang S., Cho I.K., Punyawai K., Gill B., Langmo J.N., Nath S., Greeson K.W., Symosko K.M., Fowler K.L., Tian S., Statz J.P., Steves A.N., Parnpai R., White M.A., Hennebold J.D., Orwig K.E., Simerly C.R., Schatten G., Easley C.A., IV	F and S Science
Abstract: Objective: To demonstrate that functional spermatids can be derived in vitro from nonhuman primate pluripotent stem cells. Design: Green fluorescent protein-labeled, rhesus macaque nonhuman primate embryonic stem cells (nhpESCs) were differentiated into advanced male germ cell lineages using a modified serum-free spermatogonial stem cell culture medium. In vitro-derived round spermatid-like cells (rSLCs) from differentiated nhpESCs were assessed for their ability to fertilize rhesus oocytes by intracytoplasmic sperm(atid) injection. Setting: Multiple academic laboratory settings. Patient(s): Not applicable. Intervention(s): Intracytoplasmic sperm(atid) injection of in vitro-derived spermatids from nhpESCs into rhesus macaque oocytes. Main Outcome Measure(s): Differentiation into spermatogenic cell lineages was measured through multiple assessments including ribonucleic acid sequencing and immunocytochemistry for various spermatogenic markers. In vitro spermatids were assessed for their ability to fertilize oocytes by intracytoplasmic sperm(atid) injection by assessing early fertilization events such as spermatid deoxyribonucleic acid decondensation and pronucleus formation/apposition. Preimplantation embryo development from the one-cell zygote stage to the blastocyst stage was also assessed. Result(s): Nonhuman primate embryonic stem cells can be differentiated into advanced germ cell lineages, including haploid rSLCs. These rSLCs undergo deoxyribonucleic acid decondensation and pronucleus formation/apposition when microinjected into rhesus macaque mature oocytes, which, after artificial activation and coinjection of ten-eleven translocation 3 protein, undergo embryonic divisions with approximately 12% developing successfully into expanded blastocysts. Conclusion(s): This work demonstrates that rSLCs, generated in vitro from primate pluripotent stem cells, mimic many of the capabilities of in vivo round spermatids and perform events essential for preimplantation development. To our knowledge, this work represents, for the first time, that functional spermatid-like cells can be derived in vitro from primate pluripotent stem cells. © 2021 The Author(s) Keyword: blastocysts; In vitro spermatogenesis; round spermatids; TET3 Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85118827937&doi=10.1016%2fj.xfss.2021.09.001&partnerID=40&md5=e8966a3af81b625aceea2aa0af415b83 DOI: 10.1016/j.xfss.2021.09.001
2021	CAG repeat instability in embryonic stem cells and derivative spermatogenic cells of transgenic Huntington’s disease monkey Khampang S., Parnpai R., Mahikul W., Easley C.A., IV, Cho I.K., Chan A.W.S.	Journal of Assisted Reproduction and Genetics
Abstract: Purpose: The expansion of CAG (glutamine; Q) trinucleotide repeats (TNRs) predominantly occurs through male lineage in Huntington’s disease (HD). As a result, offspring will have larger CAG repeats compared to their fathers, which causes an earlier onset of the disease called genetic anticipation. This study aims to develop a novel in vitro model to replicate CAG repeat instability in early spermatogenesis and demonstrate the biological process of genetic anticipation by using the HD stem cell model for the first time. Methods: HD rhesus monkey embryonic stem cells (rESCs) were cultured in vitro for an extended period. Male rESCs were used to derive spermatogenic cells in vitro with a 10-day differentiation. The assessment of CAG repeat instability was performed by GeneScan and curve fit analysis. Results: Spermatogenic cells derived from rESCs exhibit progressive expansion of CAG repeats with high daily expansion rates compared to the extended culture of rESCs. The expansion of CAG repeats is cell type–specific and size-dependent. Conclusions: Here, we report a novel stem cell model that replicates genome instability and CAG repeat expansion in in vitro derived HD monkey spermatogenic cells. The in vitro spermatogenic cell model opens a new opportunity for studying TNR instability and the underlying mechanism of genetic anticipation, not only in HD but also in other TNR diseases. © 2021, The Author(s), under exclusive licence to Springer Science+Business Media, LLC part of Springer Nature. Keyword: CAG repeat instability; Genetic anticipation; HD rhesus monkey embryonic stem cells (rESCs); Huntington’s disease; Spermatogenic cells Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85101087848&doi=10.1007%2fs10815-021-02106-3&partnerID=40&md5=c4249fbb7c39c2e39eb618569de706e2 DOI: 10.1007/s10815-021-02106-3
2020	Differentiation induction of human stem cells for corneal epithelial regeneration Theerakittayakorn K., Nguyen H.T., Musika J., Kunkanjanawan H., Imsoonthornruksa S., Somredngan S., Ketudat-Cairns M., Parnpai R.	International Journal of Molecular Sciences
Abstract: Deficiency of corneal epithelium causes vision impairment or blindness in severe cases. Transplantation of corneal epithelial cells is an effective treatment but the availability of the tissue source for those cells is inadequate. Stem cells can be induced to differentiate to corneal epithelial cells and used in the treatment. Multipotent stem cells (mesenchymal stem cells) and pluripotent stem cells (embryonic stem cells and induced pluripotent stem cells) are promising cells to address the problem. Various protocols have been developed to induce differentiation of the stem cells into corneal epithelial cells. The feasibility and efficacy of both human stem cells and animal stem cells have been investigated for corneal epithelium regeneration. However, some physiological aspects of animal stem cells are different from those of human stem cells, the protocols suited for animal stem cells might not be suitable for human stem cells. Therefore, in this review, only the investigations of corneal epithelial differentiation of human stem cells are taken into account. The available protocols for inducing the differentiation of human stem cells into corneal epithelial cells are gathered and compared. Also, the pathways involving in the differentiation are provided to elucidate the relevant mechanisms. © 2020 by the authors. Licensee MDPI, Basel, Switzerland. Keyword: Corneal epithelium; Differentiation; Mesenchymal stem cell; Pluripotent stem cell Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85094112207&doi=10.3390%2fijms21217834&partnerID=40&md5=23039ea71ff3873f8fbe9e92c027d7bc DOI: 10.3390/ijms21217834