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The Effect of Preferable Enrichments in the Laboratory Minipigs
Ryoung-Hoon Jeon,Seung-Joon Kim,Won-Jae Lee 한국수정란이식학회 2017 한국동물생명공학회지 Vol.32 No.4
Miniature pig (minipig) has been considered as an important laboratory animal in the developmental biotechnology researches with respect to xenotransplantation, stem cell, somatic cell nuclear transfer and embryo transfer. Given that the laboratory minipigs are normally housed at an indoor facility, they pass the time with lying or sleeping unless it is feeding time. Therefore, it is necessary to provide environmental enrichments to satisfy their innate needs and to lessen atypical behaviors caused by stress, on the purpose of welfare. We quantitatively investigated the type of preferable enrichment for the laboratory minipigs as well as its effect on their daily life. They presented a great interest to the pliable pail but a rapid loss of attraction to non-preferable enrichments. When the daily life of the single housed minipigs was quantified based on duration of playing or resting, they were more actively engaged in lively activities in the presence of enrichments. In addition, the provision of enrichments could effectively alleviate the conflicts during group housing when new pen mate was introduced, resulting in reduction of wound cases. We believe the considerations of animal welfare are essential to the conduct of better research because animals in the non-stressful environment will be more physiologically stable and provide more reliable results in the animal experiments.
Gestation length in Holstein cow depends on the number and sex of fetuses
Jeon, Ryoung-Hoon,Rho, Gyu-Jin The Korean Society of Veterinary Science 2019 大韓獸醫學會誌 Vol.59 No.4
The traditional calculation of gestation length (GL) in Holstein cows relies solely on the insemination day and has a high error rate for actual calving dates. The present study was conducted to develop a more accurate method of estimating GL. Among 3,578 Holstein cows, the number of fetuses had a significant (p < 0.05) effect, with GLs of 282.4 ± 2.5 days for singletons and 276.0 ± 1.6 days for twins. Within the singleton group, the GLs were 284.6 ± 1.9 days for males and 280.3 ± 1.5 days for females, a significant difference (p < 0.05). However, within the twin group, there was no difference in GLs among twin males, twin females, or twins of both sexes. Gestation was significantly shorter for singleton males born from June to August but significantly longer for twin males born in the May to June period (p < 0.05). Neither parity nor housing type was associated with any significant difference in GL. In order to predict the calving dates of Holstein cows accurately and improve their management, calculations should consider the number and sex of fetuses as well as the insemination season.
The Effect of Preferable Enrichments in the Laboratory Minipigs
Jeon, Ryoung-Hoon,Kim, Seung-Joon,Lee, Won-Jae The Korean Society of Embryo Transfer 2017 한국동물생명공학회지 Vol.32 No.4
Miniature pig (minipig) has been considered as an important laboratory animal in the developmental biotechnology researches with respect to xenotransplantation, stem cell, somatic cell nuclear transfer and embryo transfer. Given that the laboratory minipigs are normally housed at an indoor facility, they pass the time with lying or sleeping unless it is feeding time. Therefore, it is necessary to provide environmental enrichments to satisfy their innate needs and to lessen atypical behaviors caused by stress, on the purpose of welfare. We quantitatively investigated the type of preferable enrichment for the laboratory minipigs as well as its effect on their daily life. They presented a great interest to the pliable pail but a rapid loss of attraction to non-preferable enrichments. When the daily life of the single housed minipigs was quantified based on duration of playing or resting, they were more actively engaged in lively activities in the presence of enrichments. In addition, the provision of enrichments could effectively alleviate the conflicts during group housing when new pen mate was introduced, resulting in reduction of wound cases. We believe the considerations of animal welfare are essential to the conduct of better research because animals in the non-stressful environment will be more physiologically stable and provide more reliable results in the animal experiments.
장시정,Jeon Ryoung-Hoon,Kim Hwan-Deuk,Hwang Jong-Chan,Lee Hyeon-Jeong,배슬기,Lee Sung-Lim,노규진,김승준,Lee Won-Jae 아세아·태평양축산학회 2020 Animal Bioscience Vol.33 No.12
Objective: Quantitative polymerase chain reaction (qPCR) has been extensively used in the field of mesenchymal stem cell (MSC) research to elucidate their characteristics and clinical potential by normalization of target genes against reference genes (RGs), which are believed to be stably expressed irrespective of various experimental conditions. However, the expression of RGs is also variable depending on the experimental conditions, which may lead to false or contradictory conclusions upon normalization. Due to the current lack of information for a clear list of stable RGs in bovine MSCs, we conducted this study to identify suitable RGs in bovine MSCs. Methods: The cycle threshold values of ten traditionally used RGs (18S ribosomal RNA [18S], beta-2-microglobulin [B2M], H2A histone family, member Z [H2A], peptidylprolyl isomerase A [PPIA], ribosomal protein 4 [RPL4], succinate dehydrogenase complex, subunit A [SDHA], beta actin [ACTB], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], TATA box binding protein [TBP], and hypoxanthine phosphoribosyltrasnfrase1 [HPRT1]) in bovine bone marrow-derived MSCs (bBMMSCs) were validated for their stabilities using three types of RG evaluation algorithms (geNorm, Normfinder, and Bestkeeper). The effect of validated RGs was then verified by normalization of lineage-specific genes (fatty acid binding protein 4 [FABP4] and osteonectin [ON]) expressions during differentiations of bBMMSCs or POU class 5 homeobox 1 (OCT4) expression between bBMMSCs and dermal skins. Results: Based on the results obtained for the three most stable RGs from geNorm (TBP, RPL4, and H2A), Normfinder (TBP, RPL4, and SDHA), and Bestkeeper (TBP, RPL4, and SDHA), it was comprehensively determined that TBP and RPL4 were the most stable RGs in bBMMSCs. However, traditional RGs were suggested to be the least stable (18S) or moderately stable (GAPDH and ACTB) in bBMMSCs. Normalization of FABP4 or ON against TBP, RPL4, and 18S presented significant differences during differentiation of bBMMSCs. However, although significantly low expression of OCT4 was detected in dermal skins compared to that in bBMMSCs when TBP and RPL4 were used in normalization, normalization against 18S exhibited no significance. Conclusion: This study proposes that TBP and RPL4 were suitable as stable RGs for qPCR study in bovine MSCs.
Lee, Won-Jae,Jeon, Ryoung-Hoon,Jang, Si-Jung,Park, Ji-Sung,Lee, Seung-Chan,Baregundi Subbarao, Raghavendra,Lee, Sung-Lim,Park, Bong-Wook,King, William Allan,Rho, Gyu-Jin Hindawi Publishing Corporation 2015 Stem cells international Vol.2015 No.-
<P>The identification of stable reference genes is a prerequisite for ensuring accurate validation of gene expression, yet too little is known about stable reference genes of porcine MSCs. The present study was, therefore, conducted to assess the stability of reference genes in porcine MSCs derived from bone marrow (BMSCs), adipose (AMSCs), and skin (SMSCs) with their in vitro differentiated cells into mesenchymal lineages such as adipocytes, osteocytes, and chondrocytes. Twelve commonly used reference genes were investigated for their threshold cycle (Ct) values by qRT-PCR. The Ct values of candidate reference genes were analyzed by geNorm software to clarify stable expression regardless of experimental conditions. Thus, Pearson's correlation was applied to determine correlation between the three most stable reference genes (NF<SUB>3</SUB>) and optimal number of reference genes (NF<SUB>opt</SUB>). In assessment of stability of reference gene across experimental conditions by geNorm analysis, undifferentiated MSCs and each differentiated status into mesenchymal lineages showed slightly different results but similar patterns about more or less stable rankings. Furthermore, Pearson's correlation revealed high correlation (<I>r</I> > 0.9) between NF<SUB>3</SUB> and NF<SUB>opt</SUB>. Overall, the present study showed that<I> HMBS</I>,<I> YWHAZ</I>,<I> SDHA</I>, and<I> TBP </I>are suitable reference genes for qRT-PCR in porcine MSCs.</P>
Hyeon-Jeong Lee,Ryoung-Hoon Jeon,Byung-Joon Park,Si-Jung Jang,Sung-Lim Lee,Gyu-Jin Rho,Seung-Joon Kim,Won-Jae Lee 한국동물생명공학회(구 한국동물번식학회) 2019 Journal of Animal Reproduction and Biotechnology Vol.34 No.2
Telomeres are known as a specialized region in the end of chromosomes to protect DNA destruction, but their lengths are shortened by repetition of cell division. This telomere shortening can be preserved or be elongated by telomerase and TERT expression. Although a certain condition in the cells may affect to the cellular and molecular characteristics, the effect of differentiation induction to telomere length and telomerase activity in mesenchymal stem cells (MSCs) has been less studied. Therefore, the present study aimed to uncover periodical alterations of telomere length, telomerase activity and TERT expression in the dental pulp-derived MSCs (DP-MSCs) under condition of differentiation inductions into adipocytes and osteoblasts on a weekly basis up to 3 weeks. Shortening of telomere was significantly (p < 0.05) identified from early-middle stages of both differentiations in comparison with undifferentiated DP-MSCs by non-radioactive chemiluminescent assay and qRT-PCR method. Telomere length in undifferentiated DP-MSCs was 10.5 kb, but the late stage of differentiated DP-MSCs which can be regarded as the adult somatic cell exhibited 8.1-8.6 kb. Furthermore, the relative-quantitative telomerase repeat amplification protocol or western blotting presented significant (p < 0.05) decrease of telomerase activity since early stages of differentiations or TERT expression from middle stages of differentiations than undifferentiated state, respectively. Based on these results, it is supposed that shortened telomere length in differentiated DP-MSCs was remained along with prolonged differentiation durations, possibly due to weakened telomerase activity and TERT expression. We expect that the present study contributes on understanding differentiation mechanism of MSCs, and provides standardizing therapeutic strategies in clinical application of MSCs in the animal biotechnology.
Alteration of Apoptosis during Differentiation in Human Dental Pulp-Derived Mesenchymal Stem Cell
Hyeon-Jeong Lee,Byung-Joon Park,Ryoung-Hoon Jeon,Si-Jung Jang,Young-Bum Son,Sung-Lim Lee,Gyu-Jin Rho,Seung-Joon Kim,Won-Jae Lee 한국동물생명공학회(구 한국동물번식학회) 2019 Journal of Animal Reproduction and Biotechnology Vol.34 No.1
Because mesenchymal stem cells (MSCs) maintain distinct capacities with respect to self-renewal, differentiation ability and immunomodulatory function, they have been highly considered as the therapeutic agents for cell-based clinical application. Of particular, differentiation condition alters characteristics of MSCs, including cellular morphology, expression of gene/protein and cell surface molecule, immunological property and apoptosis. However, the previous results for differentiation-related apoptosis in MSCs have still remained controversial due to varied outcomes. Therefore, the present study aimed to disclose periodical alterations of pro- and anti-apoptosis in MSCs under differentiation inductions. The human dental pulp-derived MSCs (DP-MSCs) were differentiated into adipocytes and osteoblasts during early (1 week), middle (2 weeks) and late (3 weeks) stages, and were investigated on their apoptosis-related changes by Annexin V assay, qRT-PCR and western blotting. The ratio of apoptotic cell population was significantly (p < 0.05) elevated during the early to middle stages of differentiations but recovered up to the similar level of undifferentiated state at the late stage of differentiation. In the expression of mRNA and protein, whereas expressions of pro-apoptosis-related makers (BAX and BAK) were not altered in any kind and duration of differentiation inductions, anti-apoptosis marker (BCL2) was significantly (p < 0.05) elevated even at the early stage of differentiations. The recovery of apoptotic cell population at the late stage of differentiation is expected to be associated with the response by elevation of anti-apoptotic molecules. The present study may contribute on understanding for cellular mechanism in differentiation of MSCs and provide background data in clinical application of MSCs in the animal biotechnology to develop effective and safe therapeutic strategy.