• International Journal of Oral Biology
• /
• v.45 no.4
• /
• pp.197-203
• /
• 2020

Mesenchymal stem cells in the dental pulp exhibit a tendency for differentiation into various dental lineages and hold great potential as a major conduit for regenerative treatment in dentistry. Although they can be readily isolated from teeth, the exact characteristics of these stem cells have not been fully understood so far. When compared to two-dimensional (2D) cultures, three-dimensional (3D) cultures have the advantage of enriching the stem cell population. Hence, 3D-organoid culture and 3D-sphere culture were applied to dental pulp cells in the current study. Although the establishment of the organoid culture proved unsuccessful, the 3D-sphere culture readily initiated the stable generation of cell aggregates, which continued to grow and could be passaged to the second round. Interestingly, a significant increase in SOX2 expression was detected in the 3D-spheroid culture compared to the 2D culture. These results indicate the enrichment of the stemness-high population in the 3D-sphere culture. Thus, 3D-sphere culture may act as a link between the conventional and 3D-organoid cultures and aid in understanding the characteristics of dental pulp stem cells.

• Journal of the Korean Applied Science and Technology
• /
• v.38 no.1
• /
• pp.186-195
• /
• 2021

In this study, when stem cell culture solution is used as a cosmetic ingredient, one of the most prominent problems is that the ingredients generally have low thermal stability. Therefore, in this study, in order to find out how the stem cell culture medium is heated or preserved at high temperature, the effect of various effects of stem cells on the various effects of the stem cells was investigated. Investigated. As a result of the experiment, the wound healing assay confirmed that the cell migration increased after 6 hours, and after 24 hours, it was confirmed that the cell mobility was increased and cell division was promoted, thereby being concentrated. As a result of investigating the amount of transdermal water loss by preparing a cosmetic product containing stem cell culture solution, it was confirmed that the culture solution addition group showed an improvement rate of 31% compared to the non-added group, thereby helping in skin wound recovery. As a result of this, it is considered that this point should be considered when the stem cell culture medium is used as an active ingredient in cosmetics in the future.

• Molecules and Cells
• /
• v.37 no.6
• /
• pp.473-479
• /
• 2014

Spermatogonial stem cells (SSCs, also called germline stem cells) are self-renewing unipotent stem cells that produce differentiating germ cells in the testis. SSCs can be isolated from the testis and cultured in vitro for long-term periods in the presence of feeder cells (often mouse embryonic fibroblasts). However, the maintenance of SSC feeder culture systems is tedious because preparation of feeder cells is needed at each subculture. In this study, we developed a Matrigel-based feeder-free culture system for long-term propagation of SSCs. Although several in vitro SSC culture systems without feeder cells have been previously described, our Matrigel-based feeder-free culture system is time- and cost-effective, and preserves self-renewability of SSCs. In addition, the growth rate of SSCs cultured using our newly developed system is equivalent to that in feeder cultures. We confirmed that the feeder-free cultured SSCs expressed germ cell markers both at the mRNA and protein levels. Furthermore, the functionality of feeder-free cultured SSCs was confirmed by their transplantation into germ cell-depleted mice. These results suggest that our newly developed feeder-free culture system provides a simple approach to maintaining SSCs in vitro and studying the basic biology of SSCs, including determination of their fate.

• Anatomy and Cell Biology
• /
• v.56 no.4
• /
• pp.508-517
• /
• 2023

In cancer patients, chemo/radio therapy may cause infertility by damaging the spermatogenesis affecting the self-renewal and differentiation of spermatogonial stem cells (SSCs). In vitro differentiation of stem cells especially mesenchymal stem cells (MSCs) into germ cells has recently been proposed as a new strategy for infertility treatment. The aim of this study was to evaluate the proliferation and differentiation of SSCs using their co-culture with Sertoli cells and conditioned medium (CM) from adipose tissue-derived MSCs (AD-MSCs). Testicular tissues were separated from 2-7 days old neonate Wistar Rats and after mechanical and enzymatic digestion, the SSCs and Sertoli cells were isolated and cultured in Dulbecco's modified eagle medium with 10% fetal bovine serum, 1X antibiotic, basic fibroblast growth factor, and glial cell line-derived neurotrophic factor. The cells were treated with the CM from AD-MSCs for 12 days and then the expression level of differentiation-related genes were measured. Also, the expression level of two major spermatogenic markers of DAZL and DDX4 was calculated. Scp3, Dazl, and Prm1 were significantly increased after treatment compared to the control group, whereas no significant difference was observed in Stra8 expression. The immunocytochemistry images showed that DAZL and DDX4 were positive in experimental group comparing with control. Also, western blotting revealed that both DAZL and DDX4 had higher expression in the treated group than the control group, however, no significant difference was observed. In this study, we concluded that the CM obtained from AD-MSCs can be considered as a suitable biological material to induce the differentiation in SSCs.

• Journal of Animal Reproduction and Biotechnology
• /
• v.38 no.1
• /
• pp.26-31
• /
• 2023

Chicken embryonic stem (ES) cells have great potential and provide a powerful tool to investigate embryonic development and to manipulate genetic modification in a genome. However, very limited studies are available on the functional characterization and robust expansion of chicken ES cells compared to other species. Here, we have developed a method to generate chicken embryonic stem cell-like cells under pluripotent culture conditions. The chicken embryonic stem cell-like cells were cultivated long-term over several passages of culture without loss of pluripotency in vitro and had the specific expression of key stem cell markers. Furthermore, they showed severe changes in morphology and a significant reduction in pluripotent genes after siRNA-mediated NANOG knockdown. Collectively, these results demonstrate the efficient generation of chicken embryonic stem cell-like cells from EGK stage X blastoderm-derived singularized cells and will facilitate their potential use for various purposes, such as biobanking genetic materials and understanding stemness in the fields of animal biotechnology.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.62 no.1
• /
• pp.60-65
• /
• 2017

This study was conducted to dentify the optimum plantlet type of 'Solara' potatoes (Solanum tuberosum L.) for growth in an aeroponics system. Plantlets of 'Solara' were transplanted on March 16, 2015 in a greenhouse, and growth and yield characteristics were investigated at 70 and 78 days after transplanting, respectively. Stem length was shorter in plantlet of 15-day-old stem cuttings and acclimatization of culture, and the stem length of plantlets of stem cuttings tended to increase with increasing stem cutting age. The fresh weight of plants was the highest in the plantlets of 40-day-old stem cuttings and the lowest in non-rooted stem cuttings and acclimatization of culture. The highest number of first stolons was obtained in 35-day-old stem cuttings. The number of second stolons was the highest in plantlets of 35-day-old stem cuttings, acclimatization of culture, and 30- day-old stem cuttings. The total number of tubers was higher in plantlets of 35-day-old stem cuttings and acclimatization of culture, and the number of tubers above 3 g was the highest in plantlets of 35-day-old stem cuttings. The weight of tubers above 3 g was the heaviest in plantlets of 35-day-old stem cuttings(1,947 g per 10 plants), followed by plantlets of 30-day-old stem cuttings. These results indicate that plantlets of 30 to 35-day-old stem cuttings could be the best for production of 'Solara' potato tubers in an aeroponics system.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.50 no.2
• /
• pp.160-168
• /
• 2017

Optimizing the conditions for stem cell culture is an essential prerequisite for the efficient utilization of stem cells. In the culture of fish embryonic stem cells (ESCs) or ESC-like cells, embryo extracts are important for stable growth, but there is no rule for determining the developmental stage of the embryos used to obtain extracts. Therefore, this study investigated the effects of the developmental stage of extract donor embryos on the culture of Oryzias dancena ESC-like cells. O. dancena ESC-like cells were cultured in different media containing each of four types of embryo extract depending on the developmental stage of the extract donor embryos. Growth, morphology, colony-forming ability, alkaline phosphatase (AP) activity, and embryoid body (EB) formation of the cells were investigated. While the developmental stage of the extract donor embryos did not influence the growth, morphology, AP activity, or EB formation of ESC-like cells, colony-forming ability was affected and the pattern of the effects differed completely between the two ESC-like cells investigated. These results suggest that the developmental stage of extract donor embryos should be selected carefully for the culture of ESC-like cells, according to the research purpose and type of cell line.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.48 no.2
• /
• pp.71-78
• /
• 2022

This study was conducted to review the efficacy of different sources of stem cells in bone regeneration of cleft palate patients. The majority of previous studies focused on the transplantation of bone marrow mesenchymal stem cells. However, other sources of stem cells have also gained considerable attention, and dental stem cells have shown especially favorable outcomes. Additionally, approaches that apply the co-culture and co-transplantation of stem cells have shown promising results. The use of different types of stem cells, based on their accessibility and efficacy in bone regeneration, is a promising method in cleft palate bone regeneration. In this regard, dental stem cells may be an ideal choice due to their efficacy and accessibility. In conclusion, stem cells, despite the lengthy procedures required for culture and preparation, are a suitable alternative to conventional bone grafting techniques.

• BMB Reports
• /
• v.48 no.12
• /
• pp.668-676
• /
• 2015

Pluripotent stem cells only exist in a narrow window during early embryonic development, whereas multipotent stem cells are abundant throughout embryonic development and are retainedin various adult tissues and organs. While pluripotent stem cell lines have been established from several species, including mouse, rat, and human, it is still challenging to establish stable multipotent stem cell lines from embryonic or adult tissues. Based on current knowledge, we anticipate that by manipulating extrinsic and intrinsic signaling pathways, most if not all types of stem cells can be maintained in a long-term culture. In this article, we summarize current culture conditions established for the long-term maintenance of authentic pluripotent and multipotent stem cells and the signaling pathways involved. We also discuss the general principles of stem cell maintenance and propose several strategies on the establishment of novel stem cell lines through manipulation of signaling pathways.

• Food Science of Animal Resources
• /
• v.40 no.4
• /
• pp.659-667
• /
• 2020

Muscle stem cells isolated from domestic animals, including cows and pigs, were recently spotlighted as candidates for the production of alternative protein resources, so-called cultured meat or lab-grown meat. In the present study, we aimed to optimize the in vitro culture conditions for the long-term expansion of pig muscle stem cells via the screening of various signaling molecules. Pig muscle stem cells were collected from the biceps femoris muscles of 3-d-old crossbred pigs (Landrace×Yorkshire×Duroc, LYD) and cultured in minimum essential medium-based growth media. However, the pig muscle stem cells gradually lost their proliferation ability and featured morphologies during the long-term culture over two weeks. To find suitable in vitro culture conditions for an extended period, skeletal muscle growth medium-2, including epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580), was used to support the stemness of the pig muscle stem cells. Interestingly, pig muscle stem cells were stably maintained in a long-term culture without loss of the expression of myogenic marker genes as determined by PCR analysis. Immunostaining analysis showed that the stem cells were capable of myogenic differentiation after multiple passaging. Therefore, we found that basal culture conditions containing EGF, dexamethasone, and a p38 inhibitor were suitable for maintaining pig muscle stem cells during expanded culture in vitro. This culture method may be applied for the production of cultured meat and further basic research on muscle development in the pig.