• Molecules and Cells
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• 제23권2호
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• pp.132-137
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• 2007

With the advance of stem cell transplantation research, in vivo cell tracking techniques have become increasingly important in recent years. Magnetic resonance imaging (MRI) may provide a unique tool for non-invasive tracking of transplanted cells. Since the initial findings on the stem cell migration by MRI several years ago, there have been numerous studies using various animal models, notably in heart or brain disease models. In order to develop more reliable and clinically applicable methodologies, multiple aspects should be taken into consideration. In this review, we will summarize the current status and future perspectives of in vivo cell tracking technologies using MRI. In particular, use of different MR contrast agents and their detection methods using MRI will be described in much detail. In addition, various cell labeling methods to increase the sensitivity of signals will be extensively discussed. We will also review several key experiments, in which MRI techniques were utilized to detect the presence and/or migration of transplanted stem cells in various animal models. Finally, we will discuss the current problems and future directions of cell tracking methods using MRI.

• Asian-Australasian Journal of Animal Sciences
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• 제20권7호
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• pp.1015-1022
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• 2007

The spermatogonial stem cell (SSCs) is unique in that it is the only cell in the adult male that can contribute genes to a subsequent generation. Permanent modification of the germ cell line may be realized if stem cells could be cultured, transfected with unique genes, and then transplanted into recipient testes. We developed a culture system that supported long-term viability of SSCs. We used a retrovirus vector (pMSCV including ${\beta}$-galactosidase) to stably transfect spermatogonia following long-term culture using the system developed. Expression of the reporter gene ${\beta}$-galactosidase controlled by the retroviral vector was stable in long-term cultured SSCs. We confirmed the retroviral-mediated ${\beta}$-galactsidase gene could be expressed in germ cells in recipient mice following SSCs transplantation.

• 한국발생생물학회지:발생과생식
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• 제23권2호
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• pp.79-92
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• 2019

Humanized mice, containing engrafted human cells and tissues, are emerging as an important in vivo platform for studying human diseases. Since the development of Nod scid gamma (NSG) mice bearing mutations in the IL-2 receptor gamma chain, many investigators have used NSG mice engrafted with human hematopoietic stem cells (HSCs) to generate functional human immune systems in vivo, results in high efficacy of human cell engraftment. The development of NSG mice has allowed significant advances to be made in studies on several human diseases, including cancer and graft-versus-host-disease (GVHD), and in regenerative medicine. Based on the human HSC transplantation, organ transplantation including thymus and liver in the renal capsule has been performed. Also, immune reconstruction of cells, of the lymphoid as well as myeloid lineages, has been partly accomplished. However, crosstalk between pluripotent stem cell derived therapeutic cells with human leukocyte antigen (HLA) mis/matched types and immune CD3 T cells have not been fully addressed. To overcome this hurdle, human major histocompatibility complex (MHC) molecules, not mouse MHC molecules, are required to generate functional T cells in a humanized mouse model. Here, we briefly summarize characteristics of the humanized mouse model, focusing on development of CD3 T cells with MHC molecules. We also highlight the necessity of the humanized mouse model for the treatment of various human diseases.

• 대한약침학회지
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• 제26권4호
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• pp.357-365
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• 2023

Objectives: Since stroke is a serious health issue, novel therapeutic strategies are required. In a mouse model of ischemic stroke, this study analyzed the potential of electroacupuncture (EA) and tenuigenin (TE) to improve the efficacy of human mesenchymal stem cell (hMSC) transplantation. Methods: Middle cerebral artery occlusion (MCAO) with reperfusion was used to generate ischemic stroke. Forty-eight male C57BL/6 mice were randomly divided into five groups: control, MCAO-operated, MCAO-EA, MCAO-TE, or MCAO + EA + TE. Subsequently, hMSCs were transplanted into the ischemic region and EA, TE, or the combination was administered. Behavior assessments and immunohistochemistry were conducted to evaluate motor and cognitive recovery and hMSCs survival, migration, and differentiation. Results: The combined treatment of EA and TE exhibited enhanced hMSCs survival, migration and differentiation into neural cell lineages while suppressing astrocyte formation. Immunohistochemistry demonstrated increased neurogenesis through hMSCs transplantation in the ischemic brain. Immediate behavioral improvements were not significantly different between groups, but there was a gradual recovery in motor and cognitive function over time. Conclusion: These findings highlight the potential of EA and TE co-treatment as a therapeutic strategy for ischemic stroke, opening avenues for further research to optimize treatment protocols and elucidate underlying mechanisms.

• Clinical and Experimental Pediatrics
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• 제49권2호
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• pp.173-180
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• 2006

목 적 : CMV 감염은 여전히 조혈모세포 이식 후 가장 중요한 감염 중 하나로 이환율과 사망률의 주요 원인이다. 조혈모세포 이식 후 CMV 감염 발생에 대한 위험인자의 분석 및 CMV pp65 항원혈증에 입각한 선제치료의 효과와 질환의 경과를 평가하고자 본 연구를 시행하였다. 방 법 : 1998년 10월부터 2003년 12월까지 가톨릭대학교 성모병원 소아과에서 이식을 시행받은 환아를 대상으로 하였다. pp66항원을 이용한 항원혈증검사를 토대로 혈연간 이식 환아의 경우 CMV 항원 양성세포가 5개 이상 발견된 경우, 비혈연간 이식 환아의 경우는 CMV 항원 양성세포가 하나라도 발견된 경우 ganciclovir 선제치료를 시작하였다. 결 과 : CMV 항원혈증은 대상 환아 213명 중 88명(41.3%)에서 관찰되었고, 각각 비혈연간 골수이식(62.5%), 비혈연간 제대혈이식(36.8%), HLA-일치 혈연간 이식(25.3%)이었다. 이식유형에 따른 CMV 항원혈증 발생확률은 비혈연간 골수이식($62.5{\pm}5.4%$)이 비혈연간 제대혈이식($36.8{\pm}7.8%$) 또는 HLA-일치 혈연간 이식($25.3{\pm}4.5%$)보다 통계적으로 유의하게 높았다. 단변량 분석에 의하면 비혈연간 이식, 이식 시 환자 연령(5세 이상), 이식 전 환자의 CMV-IgG, 전처치로 전신방사선조사의 사용 및 2도 이상의 급성 이식편대 숙주병의 발생이 CMV 항원혈증 발생의 위험인자이었다. 다변량분석에 의하면 비혈연간 이식, 이식전 환자의 CMV-IgG 양성상태 및 2도 이상의 급성 이식편대 숙주병의 발생이 독립적인 위험인자이었다. 이식환자 213명 중 7례(3.3%)에서 CMV 질환이 발생하였다(고항원혈증에서 6례 발생). 결 론 : 소아 조혈모세포 이식에 있어서 CMV 감염의 위험인자는 이식 전 환자의 CMV 혈청학적 상태, 조혈모세포 공급원, 급성 이식편대 숙주병이었으며, CMV 항원혈증에 입각한 ganciclovir 선제치료는 CMV 질환의 발생을 예방하는데 효과적이었다.

• IMMUNE NETWORK
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• 제11권6호
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• pp.368-375
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• 2011

Background: Recent clinical observation reported that there was a significant correlation between change in circulating vascular endothelial growth factor (VEGF) levels and the occurrence of severe acute graft-versus-host disease (GVHD) following allogeneic hematopoietic stem cell transplantation (allo-HSCT), but the action mechanisms of VEGF in GVHD have not been demonstrated. Methods: This study investigated whether or not blockade of VEGF has an effect on acute GVHD in a lethally irradiated murine allo-HSCT model of $B6\;(H-2^b)\;{\rightarrow}B6D2F1\;(H-2^{b/d})$. Syngeneic or allogeneic recipient mice were injected subcutaneously with anti-VEGF peptides, dRK6 ($50{\mu}g/dose$) or control diluent every other day for 2 weeks (total 7 doses). Results: Administration of the dRK6 peptide after allo-HSCT significantly reduced survival with greaterclinical GVHD scores and body weight loss. Allogeneic recipients injected with the dRK6 peptide exhibited significantly increased circulating levels of VEGF and expansion of donor $CD3^+$ T cells on day +7 compared to control treated animals. The donor $CD4^+$ and $CD8^+$ T-cell subsets have differential expansion caused by the dRK6 injection. The circulating VEGF levels were reduced on day +14 regardless of blockade of VEGF. Conclusion: Together these findings demonstrate that the allo-reactive responses after allo-HSCT are exaggerated by the blockade of VEGF. VEGF seems to be consumed during the progression of acute GVHD in this murine allo-HSCT model.

• 생명과학회지
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• 제12권3호
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• pp.306-316
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• 2002

흰쥐의 좌골신경을 절단한 후 혈관내에서 배양한 신경줄기세포를 이식하여 말초신경에서도 수초의 재생이 일어나는지를 형태 학적으로 규명하고 배양한 신경줄기 세포들로부터 분화한 Schwann cell들이 회복할 수 있는지를 조사하여 다음과 같은 결론을 얻었다. 이식한 20일 후 동맥내 배양한 신경줄기세포는 Schwann cell로 분화하여 신경섬유의 재생이 일어나기 시작하였다. Schwann cell은 증식 후 재수초화를 형성하기 위하여 다른 Schwann cell들로부터 여러 가지를 자극을 받고 있었으며 NGF 소견으로 볼 때 신경외막으로부터 기존의 Schwann cell로부터 신경줄기세포의 분화가 유도되었으며 PCNA 반응으로 볼 때도 기존의 신경섬유의 Schwann cell주위에서부터 증식이 일어났다. 미세구조적으로는 Schwann cell의 재수초화, 축삭내 사립체와 미세소관의 수의 증가를 관찰할 수 있었다.

• Anatomy and Cell Biology
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• 제57권3호
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• pp.333-345
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• 2024

Stem cells transplantation (SCT) is known as a newfound strategy for multiple sclerosis (MS) treatment. Human umbilical cord mesenchymal stem cells (hUCMSCs) contain various regenerative features. Experimental autoimmune encephalomyelitis (EAE) is a laboratory model of MS. This meta-analysis study was conducted to assess the overall therapeutic effects of hUCMSCs on reduction of clinical score (CS) and restoration of active movement in EAE-induced animals. For comprehensive searching (in various English and Persian databases until May 1, 2024), the main keywords of "Experimental Autoimmune Encephalomyelitis", "Multiple Sclerosis", "Human", "Umbilical Cord", "Mesenchymal", and "Stem Cell" were hired. Collected data were transferred to the citation manager software (EndNote x8) and duplicate papers were merged. Primary and secondary screenings were applied (according to the inclusion and exclusion criteria) and eligible studies were prepared for data collection. CS of two phases of peak and recovery of EAE were extracted as the difference in means and various analyses including heterogeneity, publication bias, funnel plot, and sensitivity index were reported. Meta-analysis was applied by CMA software (v.2), P<0.05 was considered a significant level, and the confidence interval (CI) was determined 95% (95% CI). Six eligible high-quality (approved by ARRIVE checklist) papers were gathered. The difference in means of peak and recovery phases were -0.775 (-1.325 to -0.225; P=0.006; I²=90.417%) and -1.230 (-1.759 to -0.700; P<0.001; I²=93.402%), respectively. The overall therapeutic effects of SCT of hUCMSCs on the EAE cases was -1.011 (95% CI=-1.392 to -0.629; P=0.001). hUCMSCs transplantation through the intravenous route to the animal MS model (EAE) seems a considerably effective procedure for the alleviation of motor defects in both phases of peak and recovery.

• Molecules and Cells
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• 제37권6호
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• pp.473-479
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• 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.

• International Journal of Stem Cells
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• 제9권2호
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• pp.186-191
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• 2016

Background and Objectives: With the global demand for dairy protein for consumption growing annually, there has been increasing activity in the research field of dairy protein synthesis and production. From a manipulation perspective, it is more difficult to use live cattle for laboratory studies on the production of milk as well as of dairy protein such as casein, as compared with using laboratory animals like rodents. Therefore, we aimed to develop a mouse model of bovine mammary alveolar ducts for laboratory-scale studies. We studied the formation of the bovine mammary gland ductal structure by transplanting the MAC-T bovine alveolar cell line into mice. Methods and Results: MAC-T cells ($1{\times}10^7$) were suspended in Matrigel and injected into the dorsal tissue of 8-week-old male BALB/C nude mice. Histological analysis of tissue dissected from the MAC-T cell-transplanted mice after 6 weeks showed the typical morphology of the tubuloalveolar female gland, as well as glands made up of branching ducts that were surrounded by smooth muscle with small alveoli budding off the ducts. In addition, the epithelial markers CK14 and CK18 were expressed within the duct-like structure. Prolactin was detected in the duct interior in these CK14+ and CK18+ cells but not in the non-transplanted MAC-T cells. Conclusions: These results showed that duct-like tissue had been successfully formed after 6 weeks of transplantation of the CK14+ and CK18+ MAC-T cells into mice dorsal tissue. This mouse model will be a useful tool for further research on the bovine mammary gland.