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SSRN 2023年

作者： Le, Ba Anh My Nguyen, Lien Boi Linh Lam, Do Truc Phuong Lam, Chi Thien Nguyen, Nhat-Thinh Nguyen, Van Thuan Bui, Hong-Thuy Cellular Reprogramming Laboratory School of Biotechnology International University Ho Chi Minh City Viet Nam Vietnam National University Ho Chi Minh City Viet Nam

Hatched blastocyst is crucial for research on the peri-implantation period, and stem cell development, however, hatching mechanobiology still needs to be better understood due to the lack of experimental techniques. Our research aimed to use some modulating agents and timelapse systems to study hatching mechanics in late-stage porcine embryos. Fetal bovine serum (FBS), amino acids (AA), and insulin–transferrin–selenium (ITS) was singly or blended added to in vitro culture to assess their potential on developmental competence of porcine embryos. We found that addition of AA increased expansion and required supply from zygote-stage, ITS impacted detrimentally on porcine embryos. In the presence of FBS, ITS could replace AA in culture medium, and FBS solely was unfulfilled to support growth of late-stage embryos. Combination of FBS, AA and ITS was sufficient to induce an early hatching, improve quality in terms of Trophectoderm (TE) and Inner cell mass (ICM) cells, and extend culture periods of porcine hatched blastocyst. FBS supportively affected hatching dynamics in porcine blastocyst by promoting blastocoel expansion, increasing blastomere numbers in same scale of embryo, reducing hatching size and hatching completion time. Collapse and re-expansion analysis showed that FBS did not modify autonomous hydraulic control, while Mitomycin treatment in blastocyst suggested FBS induced expansion by other mechanisms rather than increase in cell numbers. Finally, for the establishment of ESC (embryonic stem cells), the result showed ICM isolated from day 7 hatched blastocyst derived in the combination of FBS, AA and ITS yielded higher efficiency in attachment, colony formation, and maintenance than those derived from blastocyst day 8. Therefore, our study demonstrated that early hatched blastocyst at day 7 might be more effective for the establishment of porcine ESCs. In conclusion, combination of FBS, AA and ITS was crucial for late-stage porcine embryo as well as derivatio

关键词： Insulin

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Treatment with Lactobacillus paracasei L30 extract induces osteogenic differentiation of human bone marrow mesenchymal stem cells in vitro

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Biomedicine and Pharmacotherapy 2025年 184卷 117913-117913页

作者： Kim, Inwook Park, Sankyu Kim, Jieun Park, So Young Seo, Jeongmin Roh, Sangho Biomedical Research Institute NeoRegen Biotech Co. Ltd. Seocho-gu 06663 South Korea Cellular Reprogramming and Embryo Biotechnology Laboratory Dental Research Institute Seoul National University School of Dentistry Seoul 08826 South Korea

Bone-related diseases such as osteoporosis pose a significant health economic burden to countries around the world and, because current treatments are insufficient, more effective therapies are desperately needed. This study explored the potential of Lactobacillus paracasei L30 extract to influence the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs). Our results showed that L30 extract significantly enhanced the expression of osteogenic markers in hBM-MSCs, including alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), and collagen type I alpha 1 (COL1A1). Mechanistic studies revealed that L30 extract activated the p38 MAPK and AKT signaling pathways, leading to phosphorylation of Glycogen synthase kinase-3 beta (GSK3β) and subsequent nuclear translocation of β-catenin. Conversely, inhibition of p38 MAPK, AKT, or knockdown of β-catenin significantly attenuated the osteogenic effects of L30 extract on hBM-MSCs. Furthermore, we found that L30 extract promoted osteogenic differentiation in primary osteoblast precursors isolated from mouse calvaria and enhances bone formation in ex vivo calvarial organ cultures. Therefore, the application of Lactobacillus paracasei L30 extract in such contexts could serve as a therapeutic approach for promoting bone formation. Collectively, our findings suggest a novel approach for the clinical management of bone-related disorders, with possible applications for treating diseases such as osteoporosis. © 2025 The Authors

关键词： GSK3β Human bone marrow mesenchymal stem cells (hBM-MSCs) Lactobacillus paracasei L30 Osteogenic differentiation p38 MAPK β-catenin

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Histone deacetylase inhibitor corrects histone H3K9 modification in round spermatid DNA at the 2-cell stage and increases the development of ROSI embryos 6th

Histone deacetylase inhibitor corrects histone H3K9 modifica...

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6th International Conference on the Development of Biomedical Engineering in Vietnam, BME 2016

作者： Minh, N.T. Tu, N.B. Tram, N.T.T. Bui, H.-T. Van Thuan, N. Laboratory for Cellular Reprogramming School of Biotechnology International University Vietnam National University Ho Chi Minh City70000 Viet Nam

ISBN: (纸本)9789811043604

Although round spermatic injection (ROSI) can be used as an assisted reproductive technology (ART) for treatment of male infertility, the successful rate of embryos and full-term development after ROSI is significantly lower than that for intracytoplasmic sperm injection (ICSI). In this study, we succeed in improving the potential of ROSI oocytes developed to the blastocyst stage and the quality of embryos by treatment of ROSI oocytes with histone deacetylase inhibitor (HDAC) scriptaid for 6 h after injection. ROSI oocytes treated with scriptaid prolong duration of histone H3K9 demethylation in round spermatid origin DNA until the 2-cell stage embryos. In contrast, high rate of ROSI embryos with histone methylated at H3K9 in round spermatid origin DNA is observed at the 2-cell stage embryos in group of ROSI oocytes without scriptaid treatment. Interestingly, our study also found that ROSI oocytes treated with scriptaid for 6 h after injection enhanced newly synthesized mRNA levels at the 2-cells stage embryos. In conclusion, this study showed that treatment of ROSI oocytes for 6 h after injection improved histone H3K9 modifications, transcriptional activity at the 2-cell stage embryos, and finally could increase the success rate of embryo development to the morula and blastocyst stage and the quality of ROSI blastocyst. © Springer Nature Singapore Pte Ltd. 2018.

关键词： DNA

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Effect of follicular fluid and floating drop culture system on the maturation of bovine oocytes 6th

Effect of follicular fluid and floating drop culture system ...

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6th International Conference on the Development of Biomedical Engineering in Vietnam, BME 2016

作者： Phung, Doan Nguyen, Linh H. K. Ho, Thanh Nhu Co, Anh Khoa N. Bui, H.-T. Van Thuan, N. Cellular Reprogramming Laboratory School of Biotechnology International University—Vietnam National University Ho Chi Minh City70000 Viet Nam

ISBN: (纸本)9789811043604

The quality of mature oocyte plays a vital role in in vitro fertilization, ICSI or cloning. Therefore, the aim of this study was to establish an optimal in vitro maturation (IVM) system for bovine oocytes. In the first experiment, the effects of the cumulus-granulosa cells complexes (OCGs) collected from ovaries of bovine from three different slaughter houses in the south of Vietnam – Ho Chi Minh City (Vissan), Di An Town (Ut Hao), Long An province (Thanh Vinh) on in vitro maturation rate were examined. In the second experiment, the IVM medium supplemented with four different concentrations of bovine follicle fluid (BFF), 5%, 10%, 15% and 20%, was investigated. Besides, two culture methods using floating drop covered with paraffin oil or 48-wells cultured dish were performed. With the results from the first experiment, oocytes collection from different slaughter houses in the South of Vietnam, Ho Chi Minh City (Vissan), Di An Town (Ut Hao), Long An (Thanh Vinh), did not affect to the oocyte maturation rate. Results from the second experiment suggested that maturation culture medium supplemented with 15% BFF and cultured in floating drop covered with paraffin oil obtained the highest rate of oocytes reached metaphase II (60.33%). In conclusion, the optimal IVM system for bovine oocytes was established consisting of floating drop containing IVM medium supplemented with 15% BFF covered with paraffin oil. © Springer Nature Singapore Pte Ltd. 2018.

关键词： Paraffins

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Female germline stem cells: Potential applications in reproductive technology 6th

Female germline stem cells: Potential applications in reprod...

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6th International Conference on the Development of Biomedical Engineering in Vietnam, BME 2016

作者： Bui, H.-T. Van Thuan, N. Cellular Reprogramming Laboratory Department of Biotechnology School of Biotechnology International University Vietnam National University Ho Chi Minh City70000 Viet Nam

ISBN: (纸本)9789811043604

One of the most significant findings in stem cell area is the establishment of female germline stem cells (FGSCs) in the early 21st century. Besides FGSC huge contribution in supporting ovarian function and fertility of females, the ability to create transgenic animals from FGSC has high efficiency. Whether the FGSC can differentiate into mature oocytes for fertilization and complete development of the embryo is an enormous question for scientists. We successfully established pig FGSC from ovarian tissue in vitro culture. Our results support that the ovary contains a small number of undifferentiated cells with stem cell characteristics. These might remain in the adult ovary and cannot proliferate in normally because of the inhibitory factors in the ovary, but under appropriate conditions, they can undergo proliferation and be differentiated into oocyte-like cells, and provide a mechanism for the self-renewal of FGSC. © Springer Nature Singapore Pte Ltd. 2018.

关键词： Stem cells

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Improve the meiotic competence of growing porcine oocytes from preantral follicle 6th

Improve the meiotic competence of growing porcine oocytes fr...

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6th International Conference on the Development of Biomedical Engineering in Vietnam, BME 2016

作者： Hoai, L.T. Yen, N.X. Thoai, L.K. Van Thuan, N. Bui, H.-T. Cellular Reprogramming Laboratory School of Biotechnology International University Vietnam National University 6 Linh Trung Thu Duc District Hochiminh70000 Viet Nam

ISBN: (纸本)9789811043604

In this study, we evaluated the effect of in vitro growth culture on meiotic competence and fertilization ability of porcine oocytes derived from preantral follicles (2–3 mm in diameter). It is reported that these growing oocytes were not able to obtain full meiotic competence after normal maturation culture. Therefore, oocyte—granulose cell complexes (OGCs) were cultured in vitro growth for 24 h in medium supplemented with 1 mM dbcAMP, 0.01 IU/mL FSH, 0.001 mg/ml 17β Estradiol and 10 ng/ml Androstenedione;then subsequent to in vitro maturation culture for 48 h before undergoing in vitro fertilization. After in vitro maturation, the percentage of growing oocytes reaching to metaphase II was similar to fully-grown oocytes collected from large antral follicles (4–6 mm in diameter). However, the fertilization ability of growing oocytes after in vitro growth was still lower compared to that of fully-grown oocytes (16 and 37%, respectively). Thus, the growing oocytes from 2 to 3 mm follicles should be pre-culture in vitro growth before undergoing in vitro maturation. © Springer Nature Singapore Pte Ltd. 2018.

关键词： Growing oocytes In vitro fertilization (IVF) In vitro growth (IVG) In vitro maturation (IVM) Preantral follicles

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Effect of dibutyryl-camp (Dbc-AMP) and follicle stimulation hormone (FSH) on meiotic competence of porcine oocytes during in vitro maturation and fertilization 6th

Effect of dibutyryl-camp (Dbc-AMP) and follicle stimulation ...

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6th International Conference on the Development of Biomedical Engineering in Vietnam, BME 2016

作者： Thuy-Van, N.T. Duy, P.T. An, T.B. Van Thuan, N. Bui, H.T. Cellular Reprogramming Laboratory Department of Biotechnology School of Biotechnology International University Vietnam National University Ho Chi Minh City70000 Viet Nam

ISBN: (纸本)9789811043604

In order to produce good mature oocyte supporting for in vitro fertilization (IVF) and enhancing the developmental competence of embryo, dibutyryl cAMP (dbcAMP) and follicle stimulation hormone (FSH) were supplemented to the in vitro maturation (IVM) medium. This study investigated the effect of dbcAMP and FSH on meiotic competence after IVM and IVF parameter. First, we examined the concentrations (0.5 and 1.0 mM) of dbcAMP on the maturation of fully-grown porcine oocytes. There was no significant difference in maturation rate. However, cumulus expansion was remarkably higher in treatment with dbcAMP 1.0 mM than 0.5 mM and non-treatment group (P © Springer Nature Singapore Pte Ltd. 2018.

关键词： Biomedical engineering

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Advances in somatic cell reprogramming: Applications in regenerative biomedicine and agriculture 6th

Advances in somatic cell reprogramming: Applications in rege...

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6th International Conference on the Development of Biomedical Engineering in Vietnam, BME 2016

作者： Kieu Linh, N.H. Bui, H.-T. Van Thuan, N. Cellular Reprogramming Laboratory Department of Biotechnology School of Biotechnology International University Vietnam National University Ho Chi Minh City70000 Viet Nam

ISBN: (纸本)9789811043604

Dolly the sheep (Wilmut et al in Nature 385:810–813, [1]), the world’s first mammal to be cloned in 1995 from a somatic cell was the greatest scientific achievement of the 20 century, which fundamentally changed the basic knowledge of biologists for somatic cells. In 2006, Shinya Yamanaka made a groundbreaking discovery that adult somatic cells can be reprogrammed to become pluripotent cells by the introduction of four pluripotent genes into somatic cells—so-called induced pluripotent stem (iPS) cells (Takanashi and Yamanaka in Cell 385:810–813, [2]). Those discoveries have opened promising in the research and applications of genomic reprogramming for regenerative biomedicine, biopharmaceutical, stem cell therapy, bio-organ, conservation of the rare and endangered animals, development of transgenic animals for breeding new animal varieties, etc. … Currently, there are three basic methods to reprogram somatic cells into totipotent stem cells or pluripotent stem cells: the first method is somatic cell nuclear transfer (SCNT) by injection of a somatic cell into an enucleated oocyte in order to produce totipotent cell (cloned animals) (Wakayama et al in Nature 394:369–374, [3]), the second is fusion of somatic cell with embryonic stem cells, and the third method is introducing 2 to 4 pluripotent genes, Oct4, Sox2, Klf4 and c-MyC into somatic cell (Takanashi and Yamanaka in Cell 385:810–813, [2]). Recently, we found that treatment of somatic cells with germinal vesicle (GV) oocytes extracts could reprogram somatic cells to stem cells, we named these cells "gviPS" Cells (Bui et al in Development 141:2235–2244, [4]). In the four methods listed above, only SCNT method can reprogram somatic cell into full-term development (offspring), also known as cloning animals. In this presentation we will focus on the most advanced technology in the world and the latest research technologies in animal cloning and the applicability of those technologies in medicine, recombinant human pro

关键词： Stem cells

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The histone deacetylase inhibitor scriptaid improves the expression pattern of TIF1β in cloned outbred mouse embryos 6th

The histone deacetylase inhibitor scriptaid improves the exp...

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6th International Conference on the Development of Biomedical Engineering in Vietnam, BME 2016

作者： Tram, N.T.T. Linh, N. H. Kieu Minh, N.T. Bui, H.-T. Van Thuan, N. Cellular Reprogramming Laboratory Department of Biotechnology School of Biotechnology International University Vietnam National University Ho Chi Minh City70000 Viet Nam

ISBN: (纸本)9789811043604

Cloning animals by somatic cell nuclear transfer (SCNT) has been successful over the past decade;however, the success rate is still extremely low. In mice, compared with other species, the success rate for the procedure has been particularly low, and most of the inbred mouse strains have never been cloned. Recently, Van Thuan et al. indicated that treatment of SCNT mouse embryos with Scriptaid, a histone deacetylase inhibitor (HDACi), significantly improved the full-term development of mouse clones. To characterize the effect of Scriptaid on the enhancement of embryos’ development after SCNT, we investigated the expression and localization of the protein TIF1β during the preimplantation development of cloned embryos of ICR strain mice. In untreated cloned blastocysts, TIF1β was absent in the inner cell mass (ICM) cells, but it was hyperexpressed in the cytoplasm of trophectoderm (TE) cells;however, the protein was detected in both TE and ICM cells in ICSI-generated blastocysts. Here, we show that the treatment of cloned embryos with Scriptaid induced a normal expression level of TIF1β in the nucleoplasm of 2-cell embryos and ICM cells of blastocysts. The results show that the success rate of embryos in developing to the blastocyst stage via Scriptaid-treated cloning was higher than that of untreated cloned embryos. We suggest that Scriptaid corrected the expression and the location pattern of TIF1β and improved the preimplantation development of cloned ICR mouse embryos. © Springer Nature Singapore Pte Ltd. 2018.

关键词： Cloning

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In vitro growth of pig oocytes isolated from early antral follicles 6th

In vitro growth of pig oocytes isolated from early antral fo...

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6th International Conference on the Development of Biomedical Engineering in Vietnam, BME 2016

作者： Tran, N.T.B. Hoang, N.H. Linh, P.K. Van Thuan, N. Bui, H.-T. Cellular Reprogramming Laboratory Department of Biotechnology School of Biotechnology International University Vietnam National University Ho Chi Minh City70000 Viet Nam

ISBN: (纸本)9789811043604

Growing oocytes (108.2 ± 2.6 μm) from pig early antral follicles (1–1.5 mm in diameter) isolated from pig ovaries were difficult to become fully grown oocytes (122.1 ± 2.2 μm) under existing culture system. The objective of this research was to establish an optimum culture system for in vitro growth of pig oocytes. The report was hypothesized androstenedione supported growth of porcine oocytes from pre antral follicles. Growing oocytes were cultured for 4 and 5 days in two kinds of culture medium in the presence of 0.001 μg/ml oestradiol alone or both 0.001 μg/ml oestradiol and 10 ng/ml androgen. In vitro growth of oocytes was performed by using 96 well plate, 48 well plate and floating drop technique. After 5 days of culture, the proliferation of oocyte surrounding cells was increased significantly. Furthermore, oocytes reached to fully growth stage when culturing in both oestradiol and androgen with 53.4, 78.3 and 69.0% as compared to control medium with 47, 52.5 and 69.4% in 96 well plate, 48 well plate and floating drop, respectively. The result demonstrates that androstenedione supported the growth of porcine oocytes and the proliferation of surrounding cells under 48 well plate culture system. © Springer Nature Singapore Pte Ltd. 2018.

关键词： Drops

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