• Journal of Animal Reproduction and Biotechnology
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• v.36 no.4
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• pp.175-182
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• 2021

Pluripotent stem cells could self-renew and differentiate into various cells. In particular, porcine pluripotent stem cells are useful for preclinical therapy, transgenic animals, and agricultural usage. These stem cells have naïve and primed pluripotent states. Naïve pluripotent stem cells represented by mouse embryonic stem cells form chimeras after blastocyst injection. Primed pluripotent stem cells represented by mouse epiblast stem cells and human embryonic stem cells. They could not produce chimeras after blastocyst injection. Populations of embryonic stem cells are not homogenous; therefore, reporter systems are used to clarify the status of stem cells and to isolate the cells. For this reason, studies of the OCT4 reporter system have been conducted for decades. This review will discuss the naïve and primed pluripotent states and recent progress in the development of porcine OCT4 reporter systems.

• Journal of Plant Biotechnology
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• v.49 no.1
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• pp.3-14
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• 2022

Leonotis nepetifolia (L.) R. Br, commonly called dagga, klip dagga, or lion's ear, has been used to effectively treat various diseases and other health problems for a long time because of its antimicrobial, anti-inflammatory, antioxidant, and analgesic activities. Several studies have attributed these biological activities to L. nepetifolia's constituent secondary metabolites, such as alkaloids, phenolics, flavonoids, tannins, steroids, glycosides, coumarins, anthocyanins, and saponins. This review aims to examine the evidence-based ethnopharmacological uses of L. nepetifolia in the treatment of bronchial asthma, diarrhea, skin diseases, malaria, burns, cancer, diabetes mellitus, and rheumatism. However, although L. nepetifolia has great potential to treat these diseases, further isolation and identification of its therapeutic phytochemical constituents are required. In addition, the performance of its extracts and phytochemicals should be thoroughly tested in preclinical and clinical trials in order to ascertain their safety and efficacy, which will prove valuable in developing new medicines.

• IMMUNE NETWORK
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• v.22 no.5
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• pp.37.1-37.16
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• 2022

Autoimmune diseases are caused by a dysfunction of the acquired immune system. In a subset of autoimmune diseases, B cells escaping immune tolerance present autoantigen and produce cytokines and/or autoantibodies, resulting in systemic or organ-specific autoimmunity. Therefore, B cell depletion with monoclonal Abs targeting B cell lineage markers is standard care therapy for several B cell-mediated autoimmune disorders. In the last 5 years, genetically-engineered cellular immunotherapies targeting B cells have shown superior efficacy and long-term remission of B cell malignancies compared to historical clinical outcomes using B cell depletion with monoclonal Ab therapies. This has raised interest in understanding whether similar durable remission could be achieved with use of genetically-engineered cell therapies for autoimmunity. This review will focus on current human clinical trials using engineered cell therapies for B cell-associated autoimmune diseases.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.47-57
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• 2000

Objective: The effects of cryopreservation with or without coculture on the in vitro development of blastomere-biopsied 8-cell mouse embryos were investigated. This experimental study was originally designed for the setup of a preclinical mouse model for the preimplantation genetic diagnosis (PGD) in human. Methods: Eight-cell embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BL(표현불가)/CBA(표현불가)). Using micromanipulation, one to four blastomeres were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acid Tyrode's solution (ATS). A slow-freezing and rapid-thawing protocol with 1.5M dimethyl sulfoxide (DMSO) and 0.1M sucrose as cryoprotectant was used for the cryopreservation of blastomere- biopsied 8-cell mouse embryos. After thawing, embryos were cultured for 110 hours in Ham's F-10 supplemented with 0.4% bovine serum albumin (BSA). In the coculture group, embryos were cultured for 110 hours on the monolayer of Vero cells in the same medium. The blastocyst formation was recorded, and the embryos developed beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. Results: The survival rate of embryos after cryopreservation was significantly lower in the blastomere-biopsied (7/8, 6/8, 5/8, and 4/8 embryos) groups than in the non-biopsied, zona intact (ZI) group. Without the coculture, the blastocyst formation rate of embryos after cryopreservation was not significantly different among ZI, the zona drilling only (ZD), and the balstomere-biopsied groups, but it was significantly lower than in the non-cryopreserved control group. The mean number of cells in embryos beyond blastocyst stage was significantly higher in the control group ($50.2{\pm}14.0$) than in 6/8 ($26.5{\pm}6.2$), 5/8 ($25.0{\pm}5.5$), and 4/8 ($17.8{\pm}7.8$) groups. With the coculture using Vero cells, the blastocyst formation rate of embryos after cryopreservation was significantly lower in 5/8 and 4/8 groups, compared with the control, 7/8, and 6/8 groups. The mean number of cells in embryos beyond blastocyst stage was also significantly lower in 4/8 group ($25.9{\pm}10.2$), compared with the control ($50.2{\pm}14.0$), 7/8 ($56.0{\pm}22.2$), and 6/8 ($55.3{\pm}25.5$) groups. Conclusion: After cryopreservation, blastomere-biopsied mouse embryos have a significantly impaired developmental competence in vitro, but this detrimental effect might be prevented by the coculture with Vero cells in 8-cell mouse embryos biopsied one or two blastomeres. Biopsy of mouse embryos after ZD with ATS is a safe and highly efficient preclinical model for PGD of human embryos.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.1
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• pp.9-20
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• 1999

The genetic defects in human gametes and embryos can cause adverse effects on overall reproductive events. Biopsy of embryos for preimplantation genetic diagnosis (PGD) offers a new possibility of having children free of the genetic disease. In addition, advanced embryo culture method may enhance the effectiveness of embryo biopsy for the practical application of PGD. This experimental study was undertaken to evaluate the effects of coculture on the development in vitro of biopsied mouse embryos as a preclinical model for PGD of human embryos. Embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BLfemale/CBAmale). Using micromanipulation, 1, 2, 3 or 4 blastomeres of 8-cell stage embryos were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acidic Tyrode's solution (ATS). After biopsy of blastomeres, embryos were cultured in vitro for 110 hours in Ham's F-10 supplemented with 0.4% BSA or cocultured on the monolayer of Vero cells in the same medium. The frequence of blastocyst formation were recorded, and the embryos beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. There was no significant difference in the blastocyst formation between the zona intact control group and the zona drilling (ZD) only, or biopsied groups. The hatching rate of all the treatment groups except 4/8 group was significantly higher than that of control group. In all the treatment groups, there was a significant reduction in the mean cell number of embryos beyond blastocyst stage ($50.2{\pm}14.0$ in control group vs. $41.2{\pm}7.9$ in ZD, $39.3{\pm}8.8$ in 7/8, $29.7{\pm}6.4$ in 6/8, $25.1{\pm}5.7$ in 5/8, and $22.1{\pm}4.3$ in 4/8 groups, p<0.05). When the same treatments were followed by coculture with Vero cells, a similar pattern was seen in the blastocyst formation and the hatching rate. However, in all the treatment groups, there was a significant increase in the mean cell number of embryos beyond blastocyst stage with coculture, compared with the parallel groups without coculture. In the cleavage rate of biopsied blastomeres cultured for 110 hours after IVF, there was no significant difference between coculture and non-coculture groups (87.2% vs. 78.7%). However, the mean cell number of embryos developed from the biopsied blastomeres was significantly higher in coculture group ($11.5{\pm}4.7\;vs.\;5.9{\pm}1.9$, p<0.05). In conclusion, biopsy of mouse embryos after ZD with ATS is a safe and highly efficient method for PGD, and coculture with Vero cells showed a positive effect on the development in vitro of biopsied mouse embryos and blastomeres as a preclinical model for PGD of human embryos.

• Molecules and Cells
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• v.45 no.12
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• pp.896-910
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• 2022

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly transmissible and potentially fatal virus. So far, most comprehensive analyses encompassing clinical and transcriptional manifestation have concentrated on the lungs. Here, we confirmed evident signs of viral infection in the lungs and spleen of SARS-CoV-2-infected K18-hACE2 mice, which replicate the phenotype and infection symptoms in hospitalized humans. Seven days post viral detection in organs, infected mice showed decreased vital signs, leading to death. Bronchopneumonia due to infiltration of leukocytes in the lungs and reduction in the spleen lymphocyte region were observed. Transcriptome profiling implicated the meticulous regulation of distress and recovery from cytokine-mediated immunity by distinct immune cell types in a time-dependent manner. In lungs, the chemokine-driven response to viral invasion was highly elevated at 2 days post infection (dpi). In late infection, diseased lungs, post the innate immune process, showed recovery signs. The spleen established an even more immediate line of defense than the lungs, and the cytokine expression profile dropped at 7 dpi. At 5 dpi, spleen samples diverged into two distinct groups with different transcriptome profile and pathophysiology. Inhibition of consecutive host cell viral entry and massive immunoglobulin production and proteolysis inhibition seemed that one group endeavored to survive, while the other group struggled with developmental regeneration against consistent viral intrusion through the replication cycle. Our results may contribute to improved understanding of the longitudinal response to viral infection and development of potential therapeutics for hospitalized patients affected by SARS-CoV-2.

• IMMUNE NETWORK
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• v.24 no.2
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• pp.7.1-7.19
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• 2024

Viral load and the duration of viral shedding of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are important determinants of the transmission of coronavirus disease 2019. In this study, we examined the effects of viral doses on the lung and spleen of K18-hACE2 transgenic mice by temporal histological and transcriptional analyses. Approximately, 1×10⁵ plaque-forming units (PFU) of SARS-CoV-2 induced strong host responses in the lungs from 2 days post inoculation (dpi) which did not recover until the mice died, whereas responses to the virus were obvious at 5 days, recovering to the basal state by 14 dpi at 1×10² PFU. Further, flow cytometry showed that number of CD8+ T cells continuously increased in 1×10² PFU-virus-infected lungs from 2 dpi, but not in 1×10⁵ PFU-virus-infected lungs. In spleens, responses to the virus were prominent from 2 dpi, and number of B cells was significantly decreased at 1×10⁵ PFU; however, 1×1² PFU of virus induced very weak responses from 2 dpi which recovered by 10 dpi. Although the defense responses returned to normal and the mice survived, lung histology showed evidence of fibrosis, suggesting sequelae of SARS-CoV-2 infection. Our findings indicate that specific effectors of the immune response in the lung and spleen were either increased or depleted in response to doses of SARS-CoV-2. This study demonstrated that the response of local and systemic immune effectors to a viral infection varies with viral dose, which either exacerbates the severity of the infection or accelerates its elimination.

• Clinical and Experimental Reproductive Medicine
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• v.17 no.1
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• pp.45-56
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• 1990

Maternal serum ${\beta}$-specific human chorionic gonadotropin(${\beta}$-hCG) and pregnancy-specific ${\beta}_1$-glycoprotein($SP_1$) levels were determined more than one per week during 11-41 days post embryo transfer(ET) in 21 consecutive pregnancies after in vitro fertilization(IVF), which included 8 normal singleton pregnancies, 3 twin pregnancies, 4 clinical abortions, 1 ectopic pregnancy, and 5 preclinical abortions. The sensitivity of serum ${\beta}$-hCG and $SP_1$ radioimmunoassays was 3mIU/ml and 0.7ng/ml relatively. At the 7th to 8th week of gestation, ultrasonographic confirmation of fetal pole and fetal heartbeat was performed. Both serm ${\beta}$-hCG and $SP_1$ levels showed logarithmic increase, but log[$SP_1$] had more steep rising curve and had wider variation than log[${\beta}$-hCG] in normal singleton pregnancies. In 3 twin pregnancies and one ectopic pregnancy, both serum ${\beta}$-hCG and $SP_1$ levels located within the 95% confidence interval of the mean levels of 8 normal singleton pregnancies(normal range). In 2 clinical abortions which had a fetal pole without heartbeat, serum ${\beta}$-hCG level showed lower limit of the normal range or just below, but all $SP_1$ levels showed within the normal range. In other 2 clinical abortions which were diagnosed as blighted ovum, both serum ${\beta}$-hCG levels from 11 days post-ET and serum $SP_1$ levels from later days compared with ${\beta}$-hCG were below the normal range. In 5 preclinical abortions, serum $SP_1$ levels were within the normal range but serum ${\beta}$-hCG levels were far below the normal range. In conclusion, both serum ${\beta}$-hCG and $SP_1$ levels increased exponentially with similar pattern in normally conceived pregancy after IVF-ET. Both serum ${\beta}$-hCG and $SP_1$ levels could predict outcome of early pregnancy to a certain degree, but log[${\beta}$-hCG] levels had more significant correlation with outcome of pregnancy compared with log[$SP_1$] levels. In addition, ultrasonographic examination of fetal poles and fetal heartbeats gives very important clinical information and prognosis.

• Journal of Korean Medicine Rehabilitation
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• v.30 no.1
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• pp.79-93
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• 2020

Objectives While sciatic neuropathy is one of the common symptoms which have the lifetime incidence of 13~40%, still there is no consensus about the standardized and the most effective conservative treatment. In addition, the importance of systematic review and meta-analysis of preclinical are growing as they could suggest possible effective treatment strategy for future studies. Therefore, we conducted systematic review and meta-analysis to estimate analgesic effect of acupuncture on sciatica in rat models. Methods Systematic search was conducted for all controlled comparative preclinical trials which assessed analgesic effect of acupuncture in sciatica rat models. Database of PubMed, EMBASE, Web of Science, CNKI and 6 Korean databases were used. The primary outcome was pain, which is evaluated by stimulus behavior tests in rat models. We assessed the methodological quality with Systematic Review Centre for Laboratory Animal Experimentation's risk of bias tool. RevMan 5.3 was used for meta-analysis and subgroup analysis was conducted according to treatment site, acupuncture point, treatment period and frequency used in electroacupuncture. Results 14 studies were finally included following our inclusion criteria. The data from meta-analysis indicated that the acupuncture significantly improved the result values of behavior tests for pain evaluation, compared to no-treatment control group in animal models (standardized mean difference=4.43, 95% confidence interval 3.16 to 5.69, Z=6.84, p<0.00001; χ²=68.02, p<0.00001; I²=82%). The results of subgroup analysis indicate that acupuncture treatment of unilateral site, distal acupoints, longer treatment period and applying 2/100 Hz frequency in electroacupuncture could be more effective. Conclusions Systematic review and meta-analysis of animal studies are getting important for the future clinical studies and the improvement of heatlh care. Therefore the results of the study would provide evidence and better design for the forthcoming studies.

• Bulletin of the Korean Chemical Society
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• v.29 no.7
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• pp.1303-1310
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• 2008

Inhibitors of farnesyltransferase (FT), a key enzyme in the post-translational modifications of Ras proteins, have been extensively studied as novel anticancer agents in the preclinical stages, some of which are currently in clinical development. Previously, it has been reported that a novel FT inhibitor LB42907 inhibits Ras farnesylation in the nanomolar range in vitro. The aim of this study was to assess the antitumor efficacy of LB42907 in vitro and in vivo. Anchorage-independent growth of various human tumor cell lines was potently inhibited by treatment with LB42907, comparable to other FT inhibitors in clinical development. In the nude mouse, oral administration of LB42907 demonstrated potent antitumor activity in several human tumor xenograft models including bladder, lung and pancreas origin. Interestingly, significant tumor regression in EJ (bladder) and A549 (lung) xenografts was induced by LB42907 treatment. The effectiveness of LB42907 was also investigated in simultaneous combination with paclitaxel, vincristine, cisplatin or gemcitabine against NCI-H460, A549, and HCT116 cells in vitro using median-effect analysis. LB42907 markedly synergized with most anticancer drugs tested in this study in NCI-H460 cell. In contrast, LB42907 displayed antagonism or partial synergism with these drugs in A549 and HCT116 cells, depending on the class of combined drugs and/ or the level of cytotoxicity. Our results demonstrate that LB42907 is an effective antitumor agent in vitro and in vivo and combination of LB42907 with other chemotherapeutic drugs results in synergistic or antagonistic effects mainly in a cell line-dependent manner. Further preclinical study is warranted.