Xenotransplantation最新文献

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Detection of Anti-Non-α-Gal Xenoreactive Antibodies in Human Blood Products. 人血液制品中抗非α-半乳糖异反应性抗体的检测。
IF 3.3 4区 医学
Xenotransplantation Pub Date : 2025-03-01 DOI: 10.1111/xen.70034
Joseph M Ladowski, Meghan Hu, Janghoon Yoon, Zheng Chen, Stuart Knechtle, Annette M Jackson, Jean Kwun
{"title":"Detection of Anti-Non-α-Gal Xenoreactive Antibodies in Human Blood Products.","authors":"Joseph M Ladowski, Meghan Hu, Janghoon Yoon, Zheng Chen, Stuart Knechtle, Annette M Jackson, Jean Kwun","doi":"10.1111/xen.70034","DOIUrl":"10.1111/xen.70034","url":null,"abstract":"<p><strong>Background: </strong>Surgical bleeding is a risk in any solid organ transplant, and is commonly addressed with the transfusion of human blood products to replace or supplement coagulation factors. It is unknown if these blood products would harm xenotransplanted pig organs in human recipients demonstrating coagulopathy. The aim of this study was to investigate in vitro if blood products such as fresh frozen plasma (FFP) or cryoprecipitate (cryo) contain xenoantibodies capable of cytotoxicity to GTKO pig cells.</p><p><strong>Methods: </strong>We obtained 12 individual single-donor (7 FFP and 5 cryo) blood products from our institution's blood bank for testing. Peripheral blood mononuclear cells (PBMCs) were obtained from a GTKO/hCD55 pig for use as target cells. We performed a series of flow cytometry crossmatch (FCXM) and complement-dependent cytotoxicity (CDC) assays.</p><p><strong>Results: </strong>We found that all the tested blood products contained some degree of IgM and IgG xenoantibody. Tests using a 1:50 dilution revealed a significant decrease in IgM xenoantibody binding, but an increase in the detection of IgG binding. Multiple preparations were capable of GTKO PBMC cytotoxicity but the level of antibody binding and cell death varied by preparation.</p><p><strong>Conclusions: </strong>Both FFP and cryo contain IgM and IgG non-galactose-α-1,3-galactose (αGal) xenoantibodies capable of killing GTKO PBMCs, though the level varies by preparation. Although some centers utilize a genetic background with mutations in the three enzymes responsible for the known xenoantigens, others are investigating the GTKO pig as a potential option. These results suggest that a center pursuing a human xenotransplantation study with a GTKO genetic background should pre-screen blood products prior to administration.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":"32 2","pages":"e70034"},"PeriodicalIF":3.3,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143664697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Combinatorial Decellularization as a Better Approach to Porcine Liver Extracellular Matrix Scaffold Fabrication With Preserved Bioactivity: A Comparative Evaluation. 组合脱细胞是制备具有保留生物活性的猪肝细胞外基质支架的更好方法:比较评价。
IF 3.3 4区 医学
Xenotransplantation Pub Date : 2025-03-01 DOI: 10.1111/xen.70031
Jesna Puthiya Veettil, Devika Sasikumar Lolitha, Umashankar Payanam Ramachandra
{"title":"Combinatorial Decellularization as a Better Approach to Porcine Liver Extracellular Matrix Scaffold Fabrication With Preserved Bioactivity: A Comparative Evaluation.","authors":"Jesna Puthiya Veettil, Devika Sasikumar Lolitha, Umashankar Payanam Ramachandra","doi":"10.1111/xen.70031","DOIUrl":"10.1111/xen.70031","url":null,"abstract":"<p><p>Soft tissue repair patches of decellularized extracellular matrices (ECM) with inherently preserved structural components and biomacromolecules are desirable in regenerative applications. This study characterizes three detergent-based decellularization methods to fabricate acellular porcine liver matrices to remove antigenic determinants without compromising the structural integrity, glycosaminoglycans (GAG) content, and bound growth factors within the resulting ECM. Three detergents chosen for decellularization were sodium dodecyl sulfate (SDS), SDS with sodium deoxycholate (SDS+SDC-combinatorial method), and triton X-100 followed by SDS. Combinatorial detergent decellularization effectively removed cellular components and retained intact collagenous structure with minimal residual DNA and protein. It also preserved significantly higher amounts of GAG, HGF, and bFGF. TX100 decellularization was highly destructive with the least preservation of GAG and GFs. The SDS method showed an intermediate level of preservation of biomolecules. The correlation obtained between GAG and GFs revealed quantification of GAG to be an indirect way of estimating the bound GFs preserved within the ECM. In vitro experiments revealed the non-cytotoxic nature of the scaffolds. The study revealed that, among the three methods of decellularization, the ECM scaffold fabricated by combinatorial detergent decellularization is extremely promising to be used as a soft tissue repair patch with inherent bioactive molecules for scaffold-based regenerative therapies.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":"32 2","pages":"e70031"},"PeriodicalIF":3.3,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143664691","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
International Xenotransplantation Association (IXA) Position Paper on Infectious Disease Considerations in Xenotransplantation. 国际异种移植协会(IXA)关于异种移植中传染病考虑的立场文件。
IF 3.3 4区 医学
Xenotransplantation Pub Date : 2025-03-01 DOI: 10.1111/xen.70001
Jay A Fishman, Joachim Denner, Linda Scobie
{"title":"International Xenotransplantation Association (IXA) Position Paper on Infectious Disease Considerations in Xenotransplantation.","authors":"Jay A Fishman, Joachim Denner, Linda Scobie","doi":"10.1111/xen.70001","DOIUrl":"10.1111/xen.70001","url":null,"abstract":"<p><p>Clinical xenotransplantation has the potential to address shortages of human organs for patients with end-stage organ failure. Advances in genetic engineering, immunosuppressive regimens, and infectious disease diagnostics have improved prospects for clinical xenotransplantation. Management of the infectious risks posed by clinical xenotransplantation requires biosecure breeding and validated methods for microbiological surveillance of source animals and recipients. Novel infection control protocols may complement biosafety requirements. Infectious risks in xenotransplantation include both known human pathogens common to immunosuppressed organ recipients and from porcine organisms or xenozoonoses for which the clinical manifestations are less well defined and for which microbial assays and therapies are more limited. Some pig-specific organisms do not infect human cells but have systemic manifestations when active within the xenograft. The human risk posed by porcine endogenous retroviruses (PERV) is uncertain. There are no documented transmissions of PERV in humans and swine are available with inactivated genomic PERV loci. Metagenomic sequencing will complement more traditional diagnostic tools in the detection of any unknown pathogens in xenotransplantation recipients. Such data are required for the development of protocols for donor and recipient microbiological surveillance, infection control, and antimicrobial therapies that will enhance the safety of clinical xenotransplantation.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":"32 2","pages":"e70001"},"PeriodicalIF":3.3,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143804239","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
International Xenotransplantation Association (IXA) Position Paper on Kidney Xenotransplantation. 国际异种移植协会(IXA)关于肾脏异种移植的立场文件。
IF 3.3 4区 医学
Xenotransplantation Pub Date : 2025-03-01 DOI: 10.1111/xen.70003
Raphael P H Meier, Richard N Pierson, Jay A Fishman, Leo H Buhler, Rita Bottino, Joseph M Ladowski, Burcin Ekser, Eckhard Wolf, Paolo Brenner, Francesco Ierino, Muhammad Mohiuddin, David K C Cooper, Wayne J Hawthorne
{"title":"International Xenotransplantation Association (IXA) Position Paper on Kidney Xenotransplantation.","authors":"Raphael P H Meier, Richard N Pierson, Jay A Fishman, Leo H Buhler, Rita Bottino, Joseph M Ladowski, Burcin Ekser, Eckhard Wolf, Paolo Brenner, Francesco Ierino, Muhammad Mohiuddin, David K C Cooper, Wayne J Hawthorne","doi":"10.1111/xen.70003","DOIUrl":"10.1111/xen.70003","url":null,"abstract":"<p><p>Porcine kidney xenotransplantation for end-stage renal disease (ESRD) has reached the stage of clinical testing following major advances in donor pig genetic modifications and effective immunosuppressive strategies through decades of rigorous translational research. Reports of pig kidney xenograft survival beyond 1 year posttranplant in nonhuman primate (NHP) models justify optimism for its potential as an alternative to allotransplantation. In the United States, experimental transplantations of genetically engineered (GE) porcine kidneys into brain-dead subjects and a small number of ESRD patients have shown no evidence of hyperacute rejection and adequate pig kidney function for up to several months. Here we discuss pre-clinical/clinical results, infectious disease, ethical, and regulatory considerations, and propose evidence-based recommendations. For initial clinical trials in kidney xenotransplantation, we make the following recommendations: (i) transplantation with organs from a triple knockout (TKO) donor pig, preferably with added human transgenes, (ii) an immunosuppressive regimen with induction therapy to deplete T (and possibly B) cells, and maintenance therapy based on a cluster of differentiation (CD)40/CD154 co-stimulation pathway blockade, (iii) the patient should be fully acceptable as a candidate for allotransplantation but should be unlikely ever to receive an allograft. Patients aged 60-69 years (extendable to 40-75 years, if one of the criteria mentioned below is present), of blood group B or O, and with diabetes are most at risk in this regard. Other patients who could be considered are (i) those who have lost two or more previous kidney allografts from recurrent disease in the graft, (ii) those with broad human leukocyte antigen (HLA)-reactivity but no evidence of anti-pig antibodies, including swine leukocyte antigen (SLA), and (iii) those with failing vascular access. Clinical pilot studies in carefully and highly selected patients with no alternative therapy will provide the foundation upon which to base subsequent formal expanded clinical trials.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":"32 2","pages":"e70003"},"PeriodicalIF":3.3,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143804185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
International Xenotransplantation Association (IXA) Position Paper on the History, Current Status, and Regulation of Xenotransplantation. 国际异种移植协会(IXA)关于异种移植的历史、现状和监管的立场文件。
IF 3.3 4区 医学
Xenotransplantation Pub Date : 2025-03-01 DOI: 10.1111/xen.70002
Wayne J Hawthorne, Richard N Pierson, Leo Buhler, Peter J Cowan, Jay Fishman, Rita Bottino, Raphael P H Meier, Paolo Brenner, Eckhard Wolf, Emanuele Cozzi, Muhammad M Mohiuddin
{"title":"International Xenotransplantation Association (IXA) Position Paper on the History, Current Status, and Regulation of Xenotransplantation.","authors":"Wayne J Hawthorne, Richard N Pierson, Leo Buhler, Peter J Cowan, Jay Fishman, Rita Bottino, Raphael P H Meier, Paolo Brenner, Eckhard Wolf, Emanuele Cozzi, Muhammad M Mohiuddin","doi":"10.1111/xen.70002","DOIUrl":"https://doi.org/10.1111/xen.70002","url":null,"abstract":"<p><p>Recent landmark clinical translation of xenotransplantation depended upon multiple innovations by the xenotransplant community, including the introduction of a variety of source pig genetic modifications, technical innovations, and novel immunosuppressive strategies, as well as the development of ethical and regulatory frameworks to support translation to the clinic. Each organ, tissue, or cell type intended for xenotransplantation will require application-specific preclinical milestones to be met in order to predict \"success\", as measured by ethical, safe, and efficacious translation to the clinic. Based on successful pre-clinical results and emerging evidence from decedent studies and initial clinical cases, evidence-based infectious disease, ethical, and regulatory considerations are emerging, and will be the foundations for the application-specific position papers that are currently under development. Here, we describe significant landmark events focusing upon safe and efficacious results underpinned by appropriate guidance documents developed over the past three decades that enabled recent translation to the clinic for heart and kidney xenografts. These steps have been undertaken over the past three decades by the xenotransplant community specifically led by the International Xenotransplantation Association (IXA) in consultation with the Transplantation Society (TTS) and the World Health Organization (WHO) to usher xenotransplantation to the clinic.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":"32 2","pages":"e70002"},"PeriodicalIF":3.3,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143812482","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
How to Detect Porcine Endogenous Retrovirus (PERV) Infections in Patients After Transplantation of Pig Organs. 如何检测猪器官移植后患者的猪内源性逆转录病毒 (PERV)感染。
IF 3.3 4区 医学
Xenotransplantation Pub Date : 2025-01-01 DOI: 10.1111/xen.70028
Joachim Denner, Hina Jhelum, Jinzhao Ban, Ludwig Krabben, Benedikt B Kaufer
{"title":"How to Detect Porcine Endogenous Retrovirus (PERV) Infections in Patients After Transplantation of Pig Organs.","authors":"Joachim Denner, Hina Jhelum, Jinzhao Ban, Ludwig Krabben, Benedikt B Kaufer","doi":"10.1111/xen.70028","DOIUrl":"10.1111/xen.70028","url":null,"abstract":"<p><p>Porcine endogenous retroviruses (PERVs) are integrated into the genome of all pigs and can infect human cells in culture. However, no PERV infections have been reported in recipients following preclinical or clinical xenotransplantation or deliberate infection experiments. Detection of PERV infection in transplanted recipients is challenging due to microchimerism, such as the presence of pig cells containing PERV proviruses in the recipient. Based on our previous publications on PERV detection in xenotransplant recipients, particularly from the first clinical trials, we developed a comprehensive strategy to screen for PERV infections. Recipients can be monitored for increasing levels of viral genomic RNA and mRNA using real-time reverse transcriptase (RT)-PCR, which can indicate PERV expression and replication. To test this strategy, explanted pig hearts and organs from baboons after pig heart transplantation were analyzed. No PERV genomic RNA or mRNA was detected in these tissues, although both were found in PERV-producing human control cells. Screening for antibodies against PERV as indirect evidence of infection is the method of choice. Recombinant viral proteins were prepared for use in Western blot assays. Animal antisera generated through immunization with recombinant PERV proteins served as positive controls. No antibodies against PERV were detected in transplanted baboons, even though microchimerism was observed in many of the animals' organs. For effective antibody screening, at least two PERV proteins should be used as antigens.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":"32 1","pages":"e70028"},"PeriodicalIF":3.3,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11850954/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143493936","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Xenotransplantation Literature Update July-December 2024. 异种移植文献更新2024年7月- 12月。
IF 3.3 4区 医学
Xenotransplantation Pub Date : 2025-01-01 DOI: 10.1111/xen.70027
Kasra Shirini, Joseph M Ladowski, Raphael P H Meier
{"title":"Xenotransplantation Literature Update July-December 2024.","authors":"Kasra Shirini, Joseph M Ladowski, Raphael P H Meier","doi":"10.1111/xen.70027","DOIUrl":"10.1111/xen.70027","url":null,"abstract":"<p><p>This review highlights groundbreaking progress from July to December 2024, including developments in gene-edited pigs, cellular therapies, organ preservation, and transplantation techniques. Recent advancements, particularly in gene-editing technologies and immunosuppressive protocols, have brought the field closer to clinical application. While significant ethical, immunological, and societal challenges remain, this progress underscores the immense potential of xenotransplantation to revolutionize transplantation medicine.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":"32 1","pages":"e70027"},"PeriodicalIF":3.3,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11856842/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143504637","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Patients' Information Needs for Informed Consent to Participate in First-in-Human Pig Kidney Xenotransplant Clinical Trials: A Mixed Methods Study. 患者知情同意参与首次人类猪肾异种移植临床试验的信息需求:一项混合方法研究
IF 3.3 4区 医学
Xenotransplantation Pub Date : 2025-01-01 DOI: 10.1111/xen.70016
Elisa J Gordon, Michael K Gusmano, Jessica Gacki-Smith, Hannah L Brooks, Margaret M Matthews, Dahlya Manning, Joseph Leventhal, Karen J Maschke
{"title":"Patients' Information Needs for Informed Consent to Participate in First-in-Human Pig Kidney Xenotransplant Clinical Trials: A Mixed Methods Study.","authors":"Elisa J Gordon, Michael K Gusmano, Jessica Gacki-Smith, Hannah L Brooks, Margaret M Matthews, Dahlya Manning, Joseph Leventhal, Karen J Maschke","doi":"10.1111/xen.70016","DOIUrl":"10.1111/xen.70016","url":null,"abstract":"<p><strong>Background: </strong>Transplant programs preparing to initiate first-in-human pig kidney xenotransplant clinical trials must be especially careful when obtaining participants' informed consent. Little is known about the kind of information patients want for making an informed decision about trial participation.</p><p><strong>Methods: </strong>We conducted semi-structured telephone interviews with waitlisted kidney transplant patients about information needs regarding participating in a first-in-human pig kidney xenotransplant trial, which guided development of a prototype consent form. Subsequent usability testing interviews sought patient feedback on the consent form. We analyzed qualitative data by thematic analysis and quantitative data by descriptive statistics.</p><p><strong>Results: </strong>Twenty-eight patients participated in semi-structured interviews; 16 patients participated in usability testing interviews. Most interview participants were male (68%, 56%), White (54%, 56%), or Black (36%, 31%), respectively. Interview participants identified five types of information needs: (1) the potential for infection contraction and transmission; (2) risks, benefits, and impact of xenotransplant trials; (3) xenotransplant clinical trial and recipient experience; (4) clinical trial logistics; and (5) the pig and its kidney. Usability testing participants suggested adding details to the prototype. Participants' preparedness to make a decision about participating in a xenotransplant trial increased after reviewing the prototype (12.5% vs. 31.3%, n.s.).</p><p><strong>Conclusion: </strong>We identified multiple unique types of information patients desired to make informed decisions about pig kidney xenotransplant trial participation. Transplant programs initiating xenotransplant trials should be prepared to address patients' information needs to optimize informed decision-making for trial participation. The prototype consent form may support a patient-centered approach to informed consent.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":"32 1","pages":"e70016"},"PeriodicalIF":3.3,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11851052/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143493363","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Combinatorial Decellularization as a Better Approach to Porcine Liver Extracellular Matrix Scaffold Fabrication With Preserved Bioactivity: A Comparative Evaluation. 组合脱细胞是制备具有保留生物活性的猪肝细胞外基质支架的更好方法:比较评价。
IF 3.3 4区 医学
Xenotransplantation Pub Date : 2025-01-01 DOI: 10.1111/xen.70025
Jesna Puthiya Veettil, Devika Sasikumar Lolitha, Umashankar Payanam Ramachandra
{"title":"Combinatorial Decellularization as a Better Approach to Porcine Liver Extracellular Matrix Scaffold Fabrication With Preserved Bioactivity: A Comparative Evaluation.","authors":"Jesna Puthiya Veettil, Devika Sasikumar Lolitha, Umashankar Payanam Ramachandra","doi":"10.1111/xen.70025","DOIUrl":"10.1111/xen.70025","url":null,"abstract":"<p><p>Soft tissue repair patches of decellularized extracellular matrices (ECM) with inherently preserved structural components and biomacromolecules are desirable in regenerative applications. This study characterizes three detergent-based decellularization methods to fabricate acellular porcine liver matrices to remove antigenic determinants without compromising the structural integrity, glycosaminoglycans (GAG) content, and bound growth factors within the resulting ECM. Three detergents chosen for decellularization were sodium dodecyl sulfate (SDS), SDS with sodium deoxycholate (SDS + SDC-combinatorial method), and Triton X-100 followed by SDS. Combinatorial detergent decellularization effectively removed cellular components and retained intact collagenous structure with minimal residual DNA and protein. It also preserved significantly higher amounts of GAG, HGF, and bFGF. TX100 decellularization was highly destructive with the least preservation of GAG and GFs. The SDS method showed an intermediate level of preservation of biomolecules. The correlation obtained between GAG and GFs revealed quantification of GAG to be an indirect way of estimating the bound GFs preserved within the ECM. In vitro experiments revealed the noncytotoxic nature of the scaffolds. The study revealed that, among the three methods of decellularization, ECM scaffold fabricated by combinatorial detergent decellularization is extremely promising for use as a soft tissue repair patch with inherent bioactive molecules for scaffold-based regenerative therapy.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":"32 1","pages":"e70025"},"PeriodicalIF":3.3,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143442228","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Evaluation of Complement-Dependent Cytotoxicity Assays for Gene-Edited Pig-to-Human Xenotransplantation. 基因编辑猪到人异种移植中补体依赖性细胞毒性测定的评价。
IF 3.3 4区 医学
Xenotransplantation Pub Date : 2025-01-01 DOI: 10.1111/xen.70021
Hao Feng, Man Zhang, Qiangbing Xia, Jiaxiang Du, Tao Li, Song Chen, Yi Wang, Dengke Pan, Lan Zhu, Gang Chen
{"title":"Evaluation of Complement-Dependent Cytotoxicity Assays for Gene-Edited Pig-to-Human Xenotransplantation.","authors":"Hao Feng, Man Zhang, Qiangbing Xia, Jiaxiang Du, Tao Li, Song Chen, Yi Wang, Dengke Pan, Lan Zhu, Gang Chen","doi":"10.1111/xen.70021","DOIUrl":"10.1111/xen.70021","url":null,"abstract":"<p><strong>Background: </strong>Gene-edited pigs for xenotransplantation usually contain one or more transgenes encoding human complement regulatory proteins (CRPs). Because of species differences, human CRP(s) expressed in gene-edited pigs may have difficulty inhibiting the activation of exogenous rabbit complement added to a complement-dependent cytotoxicity (CDC) assay. The use of human complement instead of rabbit complement in CDC experiments may more accurately reflect the actual regulatory activity of human CRP(s).</p><p><strong>Methods: </strong>Peripheral blood mononuclear cells (PBMCs) were obtained from one GTKO pig and two GTKO/hCD55 pigs with a high or low level of hCD55 expression. After incubation of heat-inactivated normal human sera (HINHS) with porcine PBMCs, CDC levels were measured after the addition of commercial rabbit complement or human complement. In addition, a modified one-step CDC method was established using pooled normal human sera (NHS) without the addition of an exogenous complement.</p><p><strong>Results: </strong>There was no significant difference in the binding of IgM/IgG to PBMCs from the three pigs. Both rabbit and human complement-mediated a significant cytotoxic effect on GTKO pig PBMCs (98.97% vs. 82.73%). Even the high expression of hCD55 only had a very limited inhibitory effect on rabbit complement-mediated cytotoxicity (81.70% vs. 98.97%). However, regardless of whether the expression level was high or low, hCD55 had a very remarkable inhibitory effect on human complement-mediated cytotoxicity (2.94% and 23.83% vs. 82.73%; p < 0.01). Similar results were obtained using the modified one-step CDC method. In addition, the inhibitory effect of hCD55 on C3c and C5b-9 deposition on pig PBMCs was positively correlated with the expression level of hCD55.</p><p><strong>Conclusion: </strong>The use of human complement instead of rabbit complement in CDC assays can better reflect the actual cytotoxic effect of human xenoantibodies against pig PBMCs expressing human CRP(s), and thus may have potential application to gene-edited pig-to-human xenotransplantation.</p>","PeriodicalId":23866,"journal":{"name":"Xenotransplantation","volume":"32 1","pages":"e70021"},"PeriodicalIF":3.3,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143442277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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