Stem cell investigation最新文献

{"title":"Interplay between BMP2 and Notch signaling in endothelial-mesenchymal transition: implications for cardiac fibrosis.","authors":"Pavel Docshin,&nbsp;Ahmad Bairqdar,&nbsp;Anna Malashicheva","doi":"10.21037/sci-2023-019","DOIUrl":"10.21037/sci-2023-019","url":null,"abstract":"<p><strong>Background: </strong>The endothelial-to-mesenchymal transition (EndoMT) is a crucial process in cardiovascular development and disorders. Cardiac fibrosis, characterized by excessive collagen deposition, occurs in heart failure, leading to the organ remodeling. Embryonic signaling pathways such as bone morphogenetic protein 2 (BMP2) and Notch are involved in its regulation. However, the interplay between these pathways in EndoMT remains unclear.</p><p><strong>Methods: </strong>This study investigates the downstream targets of Notch and BMP2 and their effect on EndoMT markers in cardiac mesenchymal cells (CMCs) and human umbilical vein endothelial cells (HUVECs). We transduced cell cultures with vectors carrying intracellular domain of NOTCH1 (<i>NICD</i>) and/or <i>BMP2</i> and evaluated gene expression and activation of EndoMT markers.</p><p><strong>Results: </strong>The results suggest that the Notch and BMP2 signaling pathways have common downstream targets that regulate EndoMT. The activation of BMP2 and Notch is highly dependent on cell type, and co-cultivation of CMCs and HUVECs produced opposing cellular responses to target gene expression and α-smooth muscle actin (α-SMA) synthesis.</p><p><strong>Conclusions: </strong>The balance between Notch and BMP2 signaling determines the outcome of EndoMT and fibrosis in the heart. The study's findings highlight the need for further research to understand the interaction between Notch and BMP2 in the heart and develop new therapeutic strategies for treating cardiac fibrosis.</p>","PeriodicalId":21938,"journal":{"name":"Stem cell investigation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/12/03/sci-10-2023-019.PMC10570623.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41238589","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The quest for the holy grail: overcoming challenges in expanding human hematopoietic stem cells for clinical use.","authors":"Sepideh Bastani,&nbsp;Frank J T Staal,&nbsp;Kirsten Canté-Barrett","doi":"10.21037/sci-2023-016","DOIUrl":"10.21037/sci-2023-016","url":null,"abstract":"<p><p>Hematopoietic stem cell (HSC) transplantation has been the golden standard for many hematological disorders. However, the number of HSCs obtained from several sources, including umbilical cord blood (UCB), often is insufficient for transplantation. For decades, maintaining or even expanding HSCs for therapeutic purposes has been a \"holy grail\" in stem cell biology. Different methods have been proposed to improve the efficiency of cell expansion and enhance homing potential such as co-culture with stromal cells or treatment with specific agents. Recent progress has shown that this is starting to become feasible using serum-free and well-defined media. Some of these protocols to expand HSCs along with genetic modification have been successfully applied in clinical trials and some others are studied in preclinical and clinical studies. However, the main challenges regarding <i>ex vivo</i> expansion of HSCs such as limited growth potential and tendency to differentiate in culture still need improvements. Understanding the biology of blood stem cells, their niche and signaling pathways has provided possibilities to regulate cell fate decisions and manipulate cells to optimize expansion of HSCs <i>in vitro</i>. Here, we review the plethora of HSC expansion protocols that have been proposed and indicate the current state of the art for their clinical application.</p>","PeriodicalId":21938,"journal":{"name":"Stem cell investigation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/2c/18/sci-10-2023-016.PMC10345135.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9817176","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification and characterization of enhancer elements controlling cell type-specific and signalling dependent chromatin programming during hematopoietic development.","authors":"Alexander Maytum, Ben Edginton-White, Constanze Bonifer","doi":"10.21037/sci-2023-011","DOIUrl":"10.21037/sci-2023-011","url":null,"abstract":"<p><p>The development of multi-cellular organisms from a single fertilized egg requires to differentially execute the information encoded in our DNA. This complex process is regulated by the interplay of transcription factors with a chromatin environment, both of which provide the epigenetic information maintaining cell-type specific gene expression patterns. Moreover, transcription factors and their target genes form vast interacting gene regulatory networks which can be exquisitely stable. However, all developmental processes originate from pluripotent precursor cell types. The production of terminally differentiated cells from such cells, therefore, requires successive changes of cell fates, meaning that genes relevant for the next stage of differentiation must be switched on and genes not relevant anymore must be switched off. The stimulus for the change of cell fate originates from extrinsic signals which set a cascade of intracellular processes in motion that eventually terminate at the genome leading to changes in gene expression and the development of alternate gene regulatory networks. How developmental trajectories are encoded in the genome and how the interplay between intrinsic and extrinsic processes regulates development is one of the major questions in developmental biology. The development of the hematopoietic system has long served as model to understand how changes in gene regulatory networks drive the differentiation of the various blood cell types. In this review, we highlight the main signals and transcription factors and how they are integrated at the level of chromatin programming and gene expression control. We also highlight recent studies identifying the <i>cis</i>-regulatory elements such as enhancers at the global level and explain how their developmental activity is regulated by the cooperation of cell-type specific and ubiquitous transcription factors with extrinsic signals.</p>","PeriodicalId":21938,"journal":{"name":"Stem cell investigation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/93/7f/sci-10-2023-011.PMC10316067.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9799136","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exosome-based cancer vaccine for prevention of lung cancer.","authors":"Shuhan Meng, Aaron G Whitt, Bryce F Stamp, John W Eaton, Chi Li, Kavitha Yaddanapudi","doi":"10.21037/sci-2022-030","DOIUrl":"10.21037/sci-2022-030","url":null,"abstract":"<p><strong>Background: </strong>Our earlier work has shown that a unique stem cell-based vaccine that comprises of murine embryonic stem cells (ESCs) and murine fibroblasts expressing the immunostimulant granulocyte-macrophage colony stimulating factor (GM-CSF) successfully protects mice from the outgrowth of an implantable form of murine lung cancer. The use of live ESCs raises the potential risks of inducing teratomas and autoimmunity. We have attempted to improve the safety and utility of this prophylactic vaccine by employing exosomes derived from murine ESCs engineered to produce GM-CSF (ES-exo/GM-CSF vaccine).</p><p><strong>Methods: </strong>We have previously reported that ES-exo/GM-CSF immunization does protect mice from the outgrowth of an implantable form of murine lung cancer. Here, we have investigated the cancer prevention efficacy of ES-exo/GM-CSF vaccine in an experimental metastasis model of murine lung cancer, in which Lewis lung carcinoma (LLC) cells were administered into female C57BL/6 mice (8 weeks of age) through tail vein injection and subsequently LLC tumors were established in lungs.</p><p><strong>Results: </strong>Our objective is to test the anti-cancer efficacy of ES-exo/GM-CSF vaccine in a mouse model of metastatic lung cancer. Our studies indicate that vaccination of mice with ES-exo/GM-CSF vaccine inhibited the growth of metastatic lung tumors. ES-exo/GM-CSF vactionation reduced lung tumor burden from 1.86% in non-vaccinated, LLC-challenged mice to 0.036% in corresponding vacinnated mice. Importantly, control exosomes without GM-CSF failed to provide protection against metastasized pulmonary tumors. The efficacy of ES-exo/GM-CSF vaccination was associated with a decrease in the frequencies of tumor-infiltrating immunosuppressive immune cells, including T regulatory cells, myeloid derived suppressor cells (MDSCs) and tumor-associated macrophages, as well as an increase in effector cytokine production from intra-tumoral CD8⁺ T cells.</p><p><strong>Conclusions: </strong>Overall, our research provides a novel strategy for developing a cell-free prophylactic vaccine against lung tumors.</p>","PeriodicalId":21938,"journal":{"name":"Stem cell investigation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/59/bd/sci-10-2022-030.PMC9892015.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10662347","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of adeno-associated virus (AAV) gene delivery into human bone marrow stem cells (hBMSCs).","authors":"Shaomian Yao,&nbsp;Weiqiong Rong,&nbsp;Yuanying Yuan","doi":"10.21037/sci-2022-042","DOIUrl":"https://doi.org/10.21037/sci-2022-042","url":null,"abstract":"<p><strong>Background: </strong>Efficiently delivering nucleic acid into mammalian cells is essential to overexpress genes for assessing gene functions. Human bone marrow stem cells (hBMSCs) are the most studied tissue-derived stem cells. Adeno-associated viruses (AAVs) have been used to deliver DNA into hBMSCs for various purposes. Current literature reported that transduction efficiencies of up to 65% could be achieved by AAV gene delivery into hBMSCs. Further improvement of efficiency is needed and possible. This study tested a selection of AAV serotypes for high-efficient DNA delivery into hBMSCs.</p><p><strong>Methods: </strong>hBMSCs from different donors were infected with different serotypes of AAVs containing the enhanced green fluorescence protein (<i>eGFP</i>) reporter gene driven by the CMV promoter. Green fluorescence was monitored in the infected cells at five-day intervals. Cells were collected at designated time points after the infection for reverse-transcription polymerase chain reaction (RT-PCR) and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) to assess eGFP mRNA transcription.</p><p><strong>Results: </strong>The results indicated that the order of transduction efficiency of the AAV serotypes was AAV2 > AAV2.7m8 > AAV6 > AAV6.2 > AAV1 > AAV-DJ. AAV2 could achieve almost 100% transduction at the multiplicity of infection (MOI) greater than 100K. Over 90% of cells could be transduced at 20K to 50K MOI. About 80% transduction was seen at MOIs of 10K and 15K. RT-PCR analysis showed that eGFP mRNA could be detected from day 5 to day 30 post-AAV infection. The differences in the observed transduction efficiencies of the hBMSCs from different patients indicate donor-to-donor variability, and increased eGFP mRNA was generally seen after day 15 post-AAV2 infection. Maximal eGFP transcription was detected on day 30 post-infection.</p><p><strong>Conclusions: </strong>We conclude that AAV2 and AAV2.7m8 at an MOI of 100K or greater can efficiently deliver transgene into hBMSCs with up to near 100% transduction efficiency for sustained expression over one month. However, donor-to-donor variation exists in transduction efficiency and transgene expression, especially at MOIs less than 100K.</p>","PeriodicalId":21938,"journal":{"name":"Stem cell investigation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/29/49/sci-10-2022-042.PMC9905037.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10688110","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcriptomic analysis of feeder-free culture system for maintaining naïve-state pluripotency in human pluripotent stem cells.","authors":"Wataru Isono,&nbsp;Tomoyuki Kawasaki,&nbsp;Justin K Ichida,&nbsp;Kazunori Nagasaka,&nbsp;Osamu Hiraike,&nbsp;Akihiro Umezawa,&nbsp;Hidenori Akutsu","doi":"10.21037/sci-2022-043","DOIUrl":"https://doi.org/10.21037/sci-2022-043","url":null,"abstract":"<p><strong>Background: </strong>Human pluripotent stem cells (hPSCs) such as embryonic stem cells (ESCs) and induced pluripotent stem cells (PSCs) have the capacity of self-renewal and multilineage differentiation <i>in vitro</i>. Conventional hPSCs, which are in a primed state, can produce various types of differentiated cells. However, the variability in their degree of pluripotency and differentiation propensities, which is influenced by the inductive methods and culture conditions, limit their availability. Therefore, PSCs in a naïve state are a promising source of PSCs.</p><p><strong>Methods: </strong>We recently developed a culture system for naïve hPSCs using an inhibitor of the NOTCH signaling pathway and a histone H3 methyltransferase disruptor. This culture system requires feeder cells for stably maintaining the naïve hPSCs. We aimed to develop a culture system for hPSCs that could maintain pluripotency under feeder-free conditions.</p><p><strong>Results: </strong>We used two inhibitors to develop an alternative feeder-free culture system to obtain naïve hPSCs. The naïve cells underwent stable cellular proliferation and were positive for naïve stem cell markers; in addition, they could differentiate into the three germ layers. These feeder-free dome-shaped induced pluripotent stem cells (FFDS-iPSCs) have characteristics similar to that of naïve-like PSCs.</p><p><strong>Conclusions: </strong>The naive hPSCs under feeder-free conditions could ensure supply of cells for various applications in regenerative medicine and disease modeling.</p>","PeriodicalId":21938,"journal":{"name":"Stem cell investigation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/7b/c1/sci-10-2022-043.PMC10122725.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9438013","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The role of macrophages in fracture healing: a narrative review of the recent updates and therapeutic perspectives.","authors":"Bianca Braga Frade,&nbsp;Rhayra Braga Dias,&nbsp;Sara Gemini Piperni,&nbsp;Danielle Cabral Bonfim","doi":"10.21037/sci-2022-038","DOIUrl":"https://doi.org/10.21037/sci-2022-038","url":null,"abstract":"<p><strong>Objective: </strong>This review addresses the latest advances in research on the role of macrophages in fracture healing, exploring their relationship with failures in bone consolidation and the perspectives for the development of advanced and innovative therapies to promote bone regeneration.</p><p><strong>Background: </strong>The bone can fully restore its form and function after a fracture. However, the regenerative process of fracture healing is complex and is influenced by several factors, including macrophage activity. These cells have been found in the fracture site at all stages of bone regeneration, and their general depletion or the knockdown of receptors that mediate their differentiation, polarization, and/or function result in impaired fracture healing.</p><p><strong>Methods: </strong>The literature search was carried out in the PubMed database, using combinations of the keywords \"macrophage\", \"fracture healing, \"bone regeneration\", and \"bone repair\". Articles published within the last years (2017-2022) reporting evidence from <i>in vivo</i> long bone fracture healing experiments were included.</p><p><strong>Conclusions: </strong>Studies published in the last five years on the role of macrophages in fracture healing strengthened the idea that what appears to be essential when it comes to a successful consolidation is the right balance between the M1/M2 populations, which have different but complementary roles in the process. These findings opened promising new avenues for the development of several macrophage-targeted therapies, including the administration of molecules and/or biomaterials intended to regulate macrophage differentiation and polarization, the local transplantation of macrophage precursors, and the use of exosomes to deliver signaling molecules that influence macrophage activities. However, more research is still warranted to better understand the diversity of macrophage phenotypes and their specific roles in each step of fracture healing and to decipher the key molecular mechanisms involved in the <i>in vivo</i> crosstalk between macrophages and other microenvironmental cell types, such as endothelial and skeletal stem/progenitor cells.</p>","PeriodicalId":21938,"journal":{"name":"Stem cell investigation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/eb/1e/sci-10-2022-038.PMC9936163.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9314973","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Malignant clonal evolution from high proportion of monocytes in patients with aplastic anemia: a case report.","authors":"Qiuhao Fu,&nbsp;Lingling Liu,&nbsp;Yingmei Li","doi":"10.21037/sci-2022-049","DOIUrl":"https://doi.org/10.21037/sci-2022-049","url":null,"abstract":"<p><strong>Background: </strong>Aplastic anemia (AA) is a heterogeneous group of hematopoietic failure diseases, characterized mainly by immune hyperfunction, impaired immune tolerance, the hematopoietic microenvironment, and hematopoietic stem or progenitor cell deficiency. Oligoclonal hematopoiesis and clonal evolution make the disease more complicated, and extremely challenging to diagnose. After immunosuppressive therapy (IST) and granulocyte colony-stimulating factor (G-CSF) treatment, AA patients have a risk of developing acute leukemia.</p><p><strong>Case description: </strong>Here we report a patient with a relatively high proportion of monocytes, and all other tests were consistent with severe aplastic anemia (SAA). Monocytes increased rapidly after G-CSF treatment and were eventually diagnosed as hypo-hyperplastic acute monocytic leukemia 7 months later. A high proportion of monocytes may predict malignant clonal evolution in patients with AA. In combination with the literature, we recommend paying close attention to monocytes' elevation in patients with AA for clonal evolution and accurately selecting treatment options.</p><p><strong>Conclusions: </strong>The proportion of monocytes in the blood and bone marrow of AA patients should be closely monitored. Hematopoietic stem cell transplantation (HSCT) should be performed as early as possible once monocytes continue to increase or are associated with phenotypic abnormalities or genetic mutations. The unique value of this study is that although there were case reports about AA-derived acute leukemia, we suggested that an early high proportion of monocytes may predict malignant clonal evolution in patients with AA.</p>","PeriodicalId":21938,"journal":{"name":"Stem cell investigation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/f9/13/sci-10-2022-049.PMC10200712.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9569980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A selective type of autophagy to maintain glioma stem cell activity.","authors":"Khushbu Patel,&nbsp;Esperanza Arias","doi":"10.21037/sci-2022-047","DOIUrl":"https://doi.org/10.21037/sci-2022-047","url":null,"abstract":"LAMP2A","PeriodicalId":21938,"journal":{"name":"Stem cell investigation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/dd/5a/sci-10-2022-047.PMC9892014.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9251815","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Novel Human Umbilical Di-Chimeric (HUDC) cell therapy for transplantation without life-long immunosuppression.","authors":"Maria Siemionow,&nbsp;Joanna Cwykiel,&nbsp;Lucile Chambily,&nbsp;Stephanie Gacek,&nbsp;Sonia Brodowska","doi":"10.21037/sci-2023-024","DOIUrl":"https://doi.org/10.21037/sci-2023-024","url":null,"abstract":"<p><strong>Background: </strong>Cell-based therapies are promising for tolerance induction in bone marrow (BM), solid organs, and vascularized composite allotransplantation (VCA). The toxicity of bone marrow transplantation (BMT) protocols precludes this approach from routine clinical applications. To address this problem, we developed a new therapy of Human Umbilical Di-Chimeric (HUDC) cells for tolerance induction in transplantation. This study established <i>in vitro</i> characterization of the created HUDC cells.</p><p><strong>Methods: </strong>We performed sixteen <i>ex vivo</i> polyethylene glycol (PEG)-mediated fusions of human umbilical cord blood (UCB) cells from two unrelated donors. Fusion feasibility was confirmed <i>in vitro</i> by flow cytometry (FC) and confocal microscopy (CM). The HUDC cells' genotype was assessed by lymphocytotoxicity test and short tandem repeat-polymerase chain reaction (STR-PCR) analysis, phenotype by FC, viability by LIVE/DEAD^® assay, and apoptosis level by Annexin V staining. We used COMET assay to assess HUDC cells' genotoxicity after the fusion procedure. Clonogenic properties of HUDC cells were evaluated by colony forming unit (CFU) assay. Mixed lymphocyte reaction (MLR) assay assessed immunogenic and tolerogenic properties of HUDC cells.</p><p><strong>Results: </strong>We confirmed the creation of HUDC cells from two unrelated human donors of UCB cells by FC and CM. Human leukocyte antigen (HLA) class I and II typing, and STR-PCR analysis of HUDC cells confirmed the presence of alleles and loci from both unrelated UCB donors (donor chimerism: 49%±8.3%, n=4). FC confirmed the hematopoietic phenotype of HUDC cells. We confirmed high HUDC cells' viability (0.47% of dead cells) and a low apoptosis level of fused HUDC cells (15.9%) compared to positive control of PKH-stained UCB cells (20.4%) before fusion. COMET assay of HUDC cells revealed a lack of DNA damage. CFU assay confirmed clonogenic properties of HUDC cells, and MLR assay revealed a low immunogenicity of HUDC cells.</p><p><strong>Conclusions: </strong>This study confirmed creation of a novel HUDC cell line by <i>ex vivo</i> PEG-mediated fusion of UCB cells from two unrelated donors. The unique concept of creating a HUDC cell line, representing the genotype and phenotype of both, transplant donor and the recipient, introduces a promising approach for tolerance induction in BM, solid organs, and VCA transplantation.</p>","PeriodicalId":21938,"journal":{"name":"Stem cell investigation","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/29/c5/sci-10-2023-024.PMC10442563.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10063590","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}