Ampelopsis brevipedunculata (Maxim.) Trautv., also known as porcelain berry, has been used for many years as herbal folk medicine to treat liver diseases. This study was conducted to assess the influence of various factors on the rooting of hardwood stem cuttings of this plant species, collected from Jeju Island (Korea). Three experiments were conducted: (1) Stems of three diameters (≤ 4.5, > 4.5 – < 6.5, or ≥ 6.5 mm), (2) four types of rooting media [rockwool cubes, perlite, peat moss + perlite (3:1, v/v), or vermiculite] and (3) indole-3-butyric acid (IBA) at four treatment concentrations (0, 100, 200, or 400 mg·L-1). For the IBA concentration experiments, cuttings of the various diameters were inserted to a depth of 2 cm in a commercial propagation medium in 72-cell plug trays. The planted stem cuttings were initially placed on a fog bench for 16 days at 95% ± 5% relative humidity at a temperature of 25°C ± 3°C under 95% shade. The stem cuttings were then transferred to a greenhouse bench under ambient light and fed a nutrient solution on a daily basis for 2 weeks. Among the tested stem diameters, cuttings with a stem diameter ≥ 6.5 mm produced the largest stem diameter (2.6 mm) of new shoot, but without root growth. Initial stem diameter significantly affected the length of the new shoot, and the longest shoot length (67.8 mm) was observed in cuttings with an initial stem diameter of ≥ 6.5 mm. Cuttings inserted in vermiculite produced the largest diameter of new shoot (2.4 mm), whereas cuttings grown in rockwool cubes produced the shortest, stem diameter of new shoot growth (1.9 mm). No significant differences (p ≤ 0.05) were observed in the length of new shoot growth among the various rooting media and IBA concentrations. Furthermore, all tested IBA concentrations failed to induce rooting. Collectively, these results suggest that porcelain berry cuttings with an initial diameter of ≥ 6.5 mm have higher survival and bud break, but they did not produce rooting. Further studies are needed to optimize the protocol for the rooting of hardwood stem cuttings of A. brevipedunculata (Maxim.) Trautv.

Insulin/IGF signaling (IIS) regulates different physiological processes such as metabolism, trehalose level, growth, and reproduction. Four IIS components are identified from the bean pod borer, Maruca vitrata (Lepidoptera: Crambidae). RNA interference (RNAi) of insulin receptor (InR), Forkhead Box O (FOXO), Target of Rapamycin (TOR) or Akt led to ovary dysfunction. Especially, the RNAi treatment significantly reduced the stem cell division in the germarium. However, an addition of a porcine insulin stimulated the cell division. Immature diets significantly influenced on the ovarian stem cell development.

Musculoskeletal disorders including fracture, tendonitis, osteoarthritis, and laminitis are common diseases in racehorses that can cause large economic losses in the racehorse industry. Mesenchymal stem cells (MSCs) are being applied as new clinical tools for treatment of musculoskeletal disorders of racehorses. To investigate the immunomodulatory effects of stem cell therapy, we analyzed the anti- and pro-inflammatory factors in peripheral blood mononuclear cells of racehorses before and after stem cell application using quantitative real-time RT-PCR. The expression levels of pro-inflammatory factors (CCL5, IFN-γ, IL-2, and IL-18) were decreased while those of anti-inflammatory factors (TIMP-1, IL-10, TGF-β1, and VEGF) were increased significantly after application of equine adipose tissue-derived MSCs (eAD-MSCs) to racehorses with fracture. Moreover, the expression levels of pro-inflammatory factors (IL-2, IL-18, and TNF-α) were decreased while those of anti-inflammatory factors (TIMP-1, TIMP-2, IL-10, TGF-β1, and VEGF) increased significantly after stem cell application of eAD-MSCs in racehorses with tendonitis. After evaluating immunomodulatory effects of stem cell therapy on equine musculoskeletal disorders such as fracture and tendonitis, our results showed that expression levels of pro-inflammatory factors were decreased, while those of anti-inflammatory factors increased significantly after stem cell application of eAD-MSCs. These findings suggest that the healing effects of the stem cell therapy might be due to its modulation of inflammatory factors.

Mesenchymal stem cells (MSCs), which are present in all tissues, can differentiate into cells with various specific functions. Recently, cell-based therapies using MSCs have been increasing in the veterinary research and related fields. In this study, we investigated the cellular morphology, proliferating capacities, expression of cell surface markers such as CD13, CD34, CD44, CD45, CD90, and CD105, mesodermal differentiation potentials, and expression of senescence-related markers of p53, p21, and telomerase reverse transcriptase in equine adipose tissue-derived MSCs (eAD-MSCs) after cryopreservation. The eAD-MSCs were analyzed immediately and after being frozen in liquid nitrogen for 1 year (< 1 year, G1) and more than 3 years (> 3 years, G2), respectively. After cryopreservation for 1 - 3 years, G2 eAD-MSCs showed similar cellular morphology, proliferating capacities, and expression of cell surface markers when compared with G1 eAD-MSCs. Moreover, cryopreservation did not affect the adipogenic, chondrogenic, or osteogenic differentiation potentials of G1 and G2 eAD-MSCs. Collectively, cryopreservation for (or over) 3 years maintained the stem cell phenotype and differentiation potentials of eAD-MSCs. These results will be an advantage that can be effectively used for future development of cell-based therapies.

Porcine spermatogonial stem cells (SSCs) prefer three-dimensional (3D) culture systems to 2D ones for the maintenance of self-renewal. Of the many 3D culture systems, agar-based hydrogels are candidates for supporting porcine SSC self-renewal, and there are various types of agar powder that can be used. In this study, we sought to identify an agar-based 3D hydrogel system that exhibited strong efficacy in the maintenance of porcine SSC self-renewal. First, 3D hydrogels with different mechanics were prepared with various concentrations of Bacto agar, lysogeny broth (LB) agar, and agarose powder, and the 3D hydrogel with the strongest alkaline phosphatase (AP) activity and greatest increase in colony size was identified for the different types of agar powder. Second, among the porcine SSCs cultured in the different 3D hydrogels, we analyzed the colony formation, morphology, and size; AP activity; and transcription and translation of porcine SSC-related genes, and these were compared to determine the optimal 3D hydrogel system for the maintenance of porcine SSC self-renewal. We found that 0.6% (w/v) Bacto agar-, 1% (w/v) LB agar-, and 0.2% (w/v) agarose-based 3D hydrogels showed the strongest maintenance of AP activity and the most pronounced increase in colony size in the culture of porcine SSCs. Moreover, among these hydrogels, the strongest transcription and translation of porcine SSC-related genes and largest colony size were detected in porcine SSCs cultured in the 0.2% (w/v) agarose-based 3D hydrogel, whereas there were no significant differences in colony formation and morphology. These results demonstrate that the 0.2% (w/v) agarose-based 3D hydrogel can be effectively used for the maintenance of porcine SSC self-renewal.

Induced pluripotent stem cells (iPSCs) can be generated from adult cells. Somatic cells can be reprogrammed to form iPSCs by overexpressing transcription factors such as Oct4, Sox2, cMyc, and Klf4. To maintain undifferentiated state of iPSCs in vitro, cells have traditionally been maintained on mouse embryonic fibroblast feeders and passaged by enzymatic or mechanical dissociation methods. In this study, we compared the morphology and pluripotency of porcine iPSCs (piPSCs) after subsequent passaging using enzymatic and mechanical dissociation methods. Enzymatically and mechanically passaged piPSCs showed embryonic stem cell-like morphologies with compact cell adhesion and clear colony borders. In addition, alkaline phosphatase staining was positive for both enzymatically and mechanically passaged piPSCs. However, visual observation revealed that some colonies of enzymatically passaged piPSCs were spontaneously differentiated more than those of piPSCs mechanically passaged from 5 passage. Quantitative real-time RT-PCR demonstrated that enzymatically and mechanically passaged piPSCs expressed pluripotent genes such as Oct4, Sox2 and Nanog well at early passage. Immunofluorescent staining also confirmed that pluripotent markers such as Oct4, Sox2, and Nanog were positively expressed at early passage. However, expression levels of pluripotent genes in mechanically passaged piPSCs were also higher than those in enzymatically passaged piPSCs at early passage. Collectively, we found that mechanical passage method was better than enzymatic passage in terms of morphology and pluripotency of piPSCs at early passage. Further studies are needed to compare these dissociation methods with those obtained after more passages of piPSCs.

In this study, we assessed antioxidant, anti-diabetic, and anti-Alzheimer activities of Opuntia ficus-indica var. saboten (OFI) at harvest time. OFIs were cultivated December 2015~November 2016 in Jeju island. The 70% ethanol extracts of OFI were used to investigate total polyphenol and flavonoid contents, antioxidant(DPPH and ABTS radical scavenging assay), anti-diabetic(yeast α-glucosidase and rat α-glucosidase inhibition assay), and anti-Alzheimer(Acetylcholinesterase and butyrylcholinesterase inhibition assay) activities. Total polyphenol and flavonoid contents of OFIs were 17.40~23.11 μg garlic acid/mg Ex and 2.17~6.22 ug (+)-catechine/mg Ex, respectively. DPPH and ABTS radical scavenging activity of OFIs were 131.98~184.90 mg ascorbic acid(AA) eq/100 g and 63.60~101.83 mg AA eq/100 g, respectively. In the anti-diabetic and anti-Alzheimer activities, 70% ethanol extracts of OFI exhibited moderate inhibition activity, compared to control (acarbose and beberine). Total polyphenol and flavonoid contents, antioxidant, anti-diabetic, and anti-Alzheimer activities were no significant differences by season, respectively. Therefore, information on comparative biological evaluations of OFI may be a beneficial in exploring functional food and drug development.

The purpose of this study was to utilize the stem of Opuntia ficus-indica and to optimize the extraction conditions and standardize extract process for water soluble dietary fiber and solid. Extraction process was optimized by applying various conditions such as pH, ethanol concentration, extracting temperature and time. Maximum extraction yield for water soluble dietary fiber and solid were obtained using 100% water, indicating that ethanol hampered extraction. Also, extraction was unstable with higher heat, longer extraction time, and low acidic conditions. Maximum extraction of water soluble dietary fiber (27.92%) and solid (5.6 g) were obtained at 50°C, pH7, and 7 hr extraction time. Extract had antioxidant capacity at 1105.98 mg vitamin C equivalents (VCE) on ABTS assay and 118.10 mg VCE on DPPH assay.

In this study, manufacturing equipments for urushiol free raw rhus verniciflua stem(RRVS) extracts are developed which do not need for drying process and high pressure device. The three layered pressure tank and heating medium oil boiler are designed which are more efficient than the conventional ones, and the safety of the tank is assured by the structural analysis software. Finally, the RRVS extracts are prepared by water extraction technique at 100℃ using the developed equipments. In the physicochemical experiments for the RRVS extracts, an allergen such as urushiol is not detected, whereas the antioxidant such as polyphenol and flavonoid are contained.

In this study, we investigated the effect of bisphosphonate on the osteoblastic differentiation of human dental stem cells (hDPSCs). In the first experiment, we evaluated the effect of bisphosphonate on the differentiation of hDPSCs into osteoblasts by alkaline phosphatase staining after culturing hDPSCs. As a result, on day 13, the osteogenic differentiation of hDPSC was suppressed at 5 μM in clodronate and 2 μM in zolendronate. In NBP, osteogenic differentiation is more suppressed. In second experiment, cytotoxicity and proliferation test, the cell proliferation (examined by MTT assay) was more suppressed as the concentrations of zolendronate were larger than those of alendronate and clodronate. Western blotting, a third experiment, was found that AKT phosphorylation was inhibited in cell signaling proteins involved in cell proliferation inhibition and death by bisphosphonate concentration. In human dental stem cells, bisphosphonates inhibit osteoblast differentiation, and this phenomenon is clearly observed in NBPs (zolendronate), and it has been found that it is related to AKT phosphorylation of cell signaling proteins.

The estrogen-mediated effect of mesenchymal stem cells (MSCs) is a highly critical factor for the clinical application of MSCs. However, the present study is conducted on MSCs derived from adult donors, which have different physiological status with steroid hormonal changes. Therefore, we explores the important role of 17β-estradiol (E2) in MSCs derived from female and male newborn piglets (NF- and NM-pBMSCs), which are non-sexually matured donors with steroid hormones. The results revealed that in vitro treatment of MSCs with E2 improved cell proliferation, but the rates varied according to the gender of the newborn donors. Following in vitro treatment of newborn MSCs with E2, mRNA levels of Oct3/4 and Sox2 increased in both genders of MSCs and they may be correlated with both estrogen receptor α (ERα) and ERβ in NF-pBMSCs, but NM-pBMSCs were only correlated with ERα. Moreover, E2-treated NF-pBMSCs decreased in β-galactosidase activity but no influence on NM-pBMSCs. In E2-mediated differentiation capacity, E2 induced an increase in the osteogenic and chondrogenic abilities of both pBMSCs, but adipogenic ability may increased only in NF-pBMSCs. These results demonstrate that E2 could affect both genders of newborn donor-derived MSCs, but the regulatory role of E2 varies depending on gender-dependent characteristics even though the original newborn donors had not been affected by functional steroid hormones.

The purpose of this study is to confirm whether spontaneous adipocyte generation during chondrogenic induction culture affects the chondrogenic differentiation of porcine skin-derived stem cells (pSSCs). For this purpose, chondrogenic differentiation characteristics and specific marker gene expression were analyzed using cell lines showing different characteristics of spontaneous adipocyte formation. Of the four different lines of pSSCs, the pSSCs-IV line showed higher Oil red O (ORO) and glycosaminoglycan (GAG) extraction levels. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed that the levels of adipogenic markers peroxisome proliferator-activated receptor gamma 2 (PPARγ2) and adipocyte Protein 2 (aP2) mRNAs were significantly higher in pSSCs-IV than those of the other pSSC lines (P<0.05). Among three chondrogenic markers, collagen type II (Col II) and sex determining region Y-box (Sox9) mRNAs were strongly expressed in pSSCs-IV (P<0.05), but not in aggrecan (Agg), which was significantly higher in pSSCs-II (P<0.05). These results demonstrate that the spontaneous adipocyte generation during chondrogenic differentiation has a positive effect on the chondrogenesis of pSSCs. More research is needed on the correlation between adipocyte generation and cartilage formation.

Mesenchymal stem cells (MSCs) have restricted life spans in vitro and can therefore only be expanded for a limited number of cell divisions before entering a senescent state and unequivocally stopping proliferation. Several types of cell culture systems have been used for large-scale expansion of MSCs. A recent trend in cell culture has been the change from serum-use to serum-supplement media. This study was conducted to investigate the proliferative effects of vegetable resources (VR) on equine adipose tissue-derived mesenchymal stem cells (eAD-MSCs) in the absence of serum and their possible mechanisms of action. Regulation of cell cycling is a key process involved in the fate of stem cells, including renewal and differentiation. In this study, we observed that the viability of eAD-MSCs was increased significantly when treated with VR under serum-free conditions. We also observed that expression levels of cell cycling-related proteins such as p53 and p21 were decreased, and proliferating cell nuclear antigen increased significantly in response to treatment with VR in eAD-MSCs under serum-free conditions. Furthermore, expression levels of cell survival-related proteins were increased in response to treatment with VR in eAD-MSCs under serum-free conditions. Therefore, our results suggest that VR promotes proliferation of eAD-MSCs under serum-free conditions.

Type1 diabetes mellitus (DM) is generally known to be caused by destruction of insulin-producing pancreatic β cells or an immune-related problem. Polydipsia is a representative symptom of DM, and it has been reported that this condition is closely related to xerostomia and is considered that hyposalivation from the salivary gland results in this phenomenon. Although various studies have reported that induction of diabetes reduces endogenous stem cells in other organs (heart, brain etc.), diabetes-related changes in endogenous stem cells in the salivary gland have not yet been well established. Therefore, in this study, to verify the change in salivary gland stem cells after diabetes, salivary gland tissues in the control and diabetes-induced groups were processed by histochemistry (Masson’s trichrome staining) for morphological analysis, TUNEL assay for cell death, and immunohistochemistry (Ki-67 and c-Kit) for cell proliferation and maturation. Diabetes induced by STZ leads to vacuolization, apoptosis, and reduction in proliferating cells/salivary gland stem cells in salivary glands of rats. This result suggests that diabetes may be associated with reduction in salivary gland function such as degeneration and inhibition of regeneration in the salivary gland.

Mesenchymal stem cells (MSCs) are multipotent cells able to differentiate into several cell lineages, which has implications for cell therapy and reproductive biotechnologies. Although MSCs have been isolated from many species, including humans and animals, there is limited data on MSCs from large ruminants, such as bovines. In this study, we tried to isolate and characterize bovine tongue tissue-derived MSCs (boT-MSCs) by investigating phenotype morphology, performing proliferation properties, and determining cell surface marker expression patterns, self-renewal, and differentiation potentials. As a result, the boT-MSCs were successfully isolated by collagenase digestion and maintained proliferative capacity until 20 passages. Moreover, the boT-MSCs expressed pluripotency markers (OCT3/4, SOX2, and NANOG) and MSC-specific surface markers including CD44, CD90, and CD105, but not CD45 and MHC-II. The boT-MSCs could also differentiate into mesodermal (adipocyte, osteocyte, and chondrocyte) cell lineages. Our results suggest that the tongues of bovines could be used as a source of MSCs.

This study conducted a social network analysis to investigate stem cell research which was actively being studied as an alternative for the treatment of intractable diseases due to infinite proliferation and differentiation ability. Papers was extracted from the PubMed database (DB) on the subject of ‘Induced Pluripotent Stem Cells (iPS) and Embryonic Stem Cells (ES)’, and ‘Adult Stem Cell (AS) and Mesenchymal Stem Cell (MS)’. Medical Subject Headings (MeSH) term was filtrated from each area. MeSH term of iPS and ES field was 148 (international), 71 (domestic), otherwise AS and MS were 89 (international), 78 (domestic). Keyword networks were visualized using degree centrality value and the core keywords compared. There was no difference in iPS and ES field compared with the domestic and international high-ranked keywords. Gene Therapy in the international level, Liver Regeneration and the Umbilical Cord in domestic were highly centered. In AS and MS fields, Neuron was high degree centrality both. Time lagged high ranked 30 keywords in slope were different, ‘Adipose Tissue’ increased both, otherwise ‘Stem Cell Transplantation’ did domestically. Although the absolute amounts of the research papers are different, research subjects had become similar to international trends following certain time lags. On the other hand research is conducted on the specific subjects in Korea. Keyword analysis will be useful method for searching a subject to actively being studied in stem cell research area.

The purpose of this study was to investigate and analyze the total phenolic content (TPC), antioxidant activities (FRAP, ABTS, and DPPH), and antibacterial properties of lotus (Nelumbo nucifera) extracts. Lotus leaves, stems, and seed pods were extracted with deionized water at 95℃, and with 70.5% ethanol at 85℃. The TPC ranged from 8.12 to 215.12 GAE mg/g. The ethanol extract of the seed pod had the highest TPC, and the TPC of the corresponding deionized water extract was 161.45 mg/g. FRAP values ranged from 104.03 to 3,546.39 TEAC μmol/g, ABTS radical cation scavenging activities ranged from 105.11 to 3,956.94 TEAC μmol/g, and DPPH radical scavenging activities ranged from 37.29 to 2,549.46 TEAC μmol/g. EC50 values ranged from 0.26 to 9.63 mg/mL, and 0.31 to 21.21 mg/mL for ABTS and DPPH, respectively. The ethanol and deionized water extracts of the seed pod showed higher TPC and stronger antioxidant properties (FRAP, ABTS, and DPPH) than those of characteristic of the leaf extracts. The ethanol and deionized water extracts of the seed pod showed antimicrobial activity against Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa with inhibition zones of 9.0 to 14.0 mm, and the ethanol extract of the leaf showed antimicrobial activity against B. subtilis and S. aureus with inhibition zones of 9.0 and 10.0 mm, respectively. Thus, the lotus seed pod could be used to produce novel teas, and could be a potential source of therapeutic ingredients for food and medicine.

본 연구는 농산물의 미생물학적 수준에 관하여 체계적인 연구분석(systemic analysis) 기법을 이용하여 비가열 농산물의 위생안전관리를 목적으로 연구를 수행하였다. 2001년부터 2015년까지 인쇄된 관련 연구논문을 NDSL 검색 엔진을 활용하여 자료를 수집, 리뷰, 자료 정리, 분석 및 결과 정리하였다. 농산물 분류체계를 비교 조사하여 미국의 IFSAC (Interagency Food Safety Analytics Collaboration) 의 식품분류체계에 따라 국내 농산물을 분류하였다. 선정된 22건의 논문 내 89건의 데이터를 종합한 결과, 분 류체계 내에서 국내 농산물에서 총균수 수준이 높고 대장균 오염이 가장 많은 농산물은 엽경채류와 새싹·발아채 소(sprouts)로 나타났다. 단체급식에서 생채소로 소비되는 엽경채류에서 단체급식의 다소비 농산물로서 식중독과 연관성이 높은 품목인 상추, 부추, 배추에 관하여 미생물에 관한 자료 검색, 수집, 선정 및 분석 결과 최종 33편의 논문에서 미생물의 정량적 수준은 총균수 4.15~ 7.69 log CFU/ g, 대장균군 1~6.99 log CFU/g, B. cereus 0.51~3.9 log CFU/ g으로 나타났다. GAP 데이터 기반 농산물의 미생물학적 영향을 미치는 환경인자 문헌조사 결과 퇴비, 토양, 작업 자 손, 장갑이 주요 영향인자로 나타났다. 농산물의 미생 물학적 관리를 위하여 전문가 의견과 문헌조사를 통하여 GAP도입, 취약 품목의 미생물학적 분석계획, 토양으로부터 작물의 오염을 최소화 할 수 있는 작물생산방식 도입 및 실행규범 준수가 필요하다.