Cell Transplantation最新文献

Compared to primary pancreatic islets, insulinoma cell-derived 3D pseudoislets offer a more accessible, consistent, renewable, and widely applicable model system for optimization and mechanistic studies in type 1 diabetes (T1D). Here, we report a simple and efficient method for generating 3D pseudoislets from MIN6 and NIT-1 murine insulinoma cells. These pseudoislets are homogeneous in size and morphology (~150 µm), exhibit functional glucose-stimulated insulin secretion (GSIS) up to 18 days (NIT-1) enabling long-term studies, are produced in high yield [>35,000 Islet Equivalence from 30 ml culture], and are suitable for both in vitro and in vivo studies, including for encapsulation studies. To enable non-invasive longitudinal monitoring of graft survival in vivo, we transduced NIT-1 cells with green fluorescent protein-luciferase and confirmed comparable morphology, viability, and GSIS to untransduced cells in vitro. After subcutaneous implantation, we show capability to monitor graft survival in immunodeficient mice, recurrence of autoimmunity in non-obese diabetic mice, and allorejection in C57BL/6 mice. Overall, this platform provides an accessible protocol for generating high yields of 3D pseudoislets and non-invasive longitudinal monitoring of graft survival in different models offer advantages over primary islets for optimization and mechanistic studies of β cell biology, drug discovery, T1D pathogenesis and prevention, and β cell transplantation.

Adult glioblastoma (GBM) is a highly malignant tumor with a poor prognosis and high mortality rate. As versatile 3D culture systems in vitro, organoid models are emerging as a promising new tool for GBM research and combat. However, Matrigel, the most used extracellular matrix, is animal-derived with a complex composition and significant batch-to-batch variability, requiring further optimization for GBM organoid construction. Hydrogels, high-affinity polymers, have been widely employed in organoid construction for their customizable properties. In the present study, we selected and tested several commonly used hydrogel materials-hyaluronic acid methacryloyl (HAMA), chitosan methacryloyl (CSMA), and gelatin methacryloyl (GelMA)-for the construction of GBM organoids. To address the limitations of a single Matrigel, we combined Matrigel with different hydrogels and found that hydrogels influenced glioblastoma stem cells and organoid formation in distinct ways. Matrigel-HAMA (MH) promoted the formation of independent spherical clones but with a significantly lower glioblastoma stem cell (GSC) proliferation rate. GelMA-HAMA(GH) could replace Matrigel preserving the characteristics and proliferative capacity of GSCs and supported the formation of more compact spherical clones than MH did. Further experimentation with ribosomal inhibitor CX5461 and CX5461 + IFNβ indicated that GH-based GBM organoid model constituted an efficient system for GBM drug testing, discovery, and precision medicine.

Subcutaneous transplantation is garnering attention as a potential transplantation site for pluripotent stem cell-derived islet cells to address the shortage of pancreatic islet transplant donors. However, subcutaneous transplantation of cells presents challenges related to angiogenesis, which is necessary for successful islet bioproduction. This study aimed to investigate a novel method for enhancing vascularization at the transplant site and thereby promote islet engraftment using a clinically available bioabsorbable medical device. A nonabsorbable device (agarose) or a bioabsorbable device (collagen-gelatin sheet [CGS]) loaded with basic fibroblast growth factor (bFGF) was implanted subcutaneously in C57BL/6 mice. There were two other groups of mice, one of which was implanted with CGS alone, which acted as a control, and another group that was implanted with bFGF-loaded agarose rods. Subsequently, 200 islets were transplanted into the subcutaneous pre-vascularized sites. An equivalent number of islets was also transplanted into the portal vein (IPTx) to compare transplantation efficacy. Vascularization of the graft site was evaluated before and after transplantation. bFGF significantly enhanced angiogenesis in the CGS mice. The normalization rate of blood glucose levels following islet transplantation in the bFGF-loaded CGS was group comparable to that in the bFGF-loaded agarose rod and IPTx groups. The presence of islets was confirmed using single-photon emission computed tomography (SPECT/CT), histological examination. Furthermore, it was noted that blood glucose levels rapidly increased after graft removal, showing that graft function was crucial to maintain normoglycemia. Importantly, the bFGF-loaded CGS showed a high rate of engraftment. This novel bioabsorbable medical device method exhibited remarkable efficacy in enhancing subcutaneous islet engraftment, potentially paving the way for a more straightforward and less invasive approach for islet cell transplantation in future clinical applications.

Islet transplantation represents one of the most promising therapies for curing type 1 diabetes, yet it encounters significant challenges, including early islet damage due to inflammation and hypoxia, which complicate engraftment and survival within the host. It is urgent to develop new strategies to improve islet grafts survival. In this study, we developed a microporous silk fibroin scaffold loaded with liraglutide (SF-Lira). It can provide mechanical support for the islets seeded on its surface and prevent excessive aggregation. The SF-Lira scaffold significantly protected the islets from inflammatory injury, notably enhancing islet viability. In the syngeneic islet transplantation model, SF-Lira significantly improved transplantation outcomes at the epididymal fat pad (EFP) site, with a higher percentage of mice achieving and maintaining normoglycemia compared to the control. Histological analysis revealed superior graft morphology in the SF-Lira group. Our study provides new insights into the application of SF scaffold in islet transplantation and shows potential for clinical translation in extrahepatic islet transplantation.

Adaptive transfer of autologous regulatory T cells (Treg), or ex vivo-generated immunomodulatory cells, has shown promise in reducing/withdrawing immunosuppression after organ transplantation. The effect of cryopreserving such cells is still unclear. This study aims to evaluate the effects of cryopreservation on the immunomodulatory functions of peripheral blood mononuclear cells (PBMCs) with or without pretreatment with red blood cell (RBC) lysate (ACK). Human PBMCs enriched from buffy coats of healthy blood donors were treated either with ACK or phosphate-buffered saline (PBS). Thereafter, a batch of the PBS-control subset was cryopreserved with 10% dimethyl sulfoxide (DMSO) and subsequently examined for phenotype, functionality, and relative gene expression. We found that ACK-treated PBMCs exhibited higher numbers of interferon gamma (IFN-γ)-producing cells when stimulated with viral peptides (p = 0.0078), indicating that ACK treatment may improve the antigen sensitivity of memory T cells. After cryopreservation, contaminated RBCs and granulocytes, cell viability, and CD4⁺ T-cell population decreased (p = 0.0078); IL-1β expression increased; and FoxP3 expression decreased (p = 0.0312), where the Treg population remained otherwise unchanged. Enriched Tregs from both fresh and frozen PBMCs suppressed the proliferation of anti-CD3/CD28-antibody-stimulated PBMCs equally. In conclusion, the preservation of Treg function following cryopreservation highlights its potential utility in tolerance-induction trials, providing experimental flexibility and simplified logistics.

Colorectal cancer (CRC) represents a significant cause of cancer-related mortality on a global scale. It is a highly heterogeneous cancer, and the response of patients to homogeneous drug therapy varies considerably. Patient-derived tumor organoids (PDTOs) represent an optimal preclinical model for cancer research. A substantial body of evidence from numerous studies has demonstrated that PDTOs can accurately predict a patient's response to different drug treatments. This article outlines the utilization of PDTOs in the management of CRC across a range of therapeutic contexts, including postoperative adjuvant chemotherapy, palliative chemotherapy, neoadjuvant chemoradiotherapy, targeted therapy, third-line and follow-up treatment, and the treatment of elderly patients. This article delineates the manner in which PDTOs can inform therapeutic decisions at all stages of CRC, thereby assisting clinicians in selecting treatment options and reducing the risk of toxicity and resistance associated with clinical drugs. Moreover, it identifies shortcomings of existing PDTOs, including the absence of consistent criteria for assessing drug sensitivity tests, the lack of vascular and tumor microenvironment models, and the high cost of the technology. In conclusion, despite their inherent limitations, PDTOs offer several advantages, including rapid culture, a high success rate, high consistency, and high throughput, which can be employed as a personalized treatment option for CRC. The use of PDTOs in CRC allows for the prediction of responses to different treatment modalities at various stages of disease progression. This has the potential to reduce adverse drug reactions and the emergence of resistance associated with clinical drugs, facilitate evidence-based clinical decision-making, and guide CRC patients in the selection of personalized medications, thereby advancing the individualized treatment of CRC.

B-cell lymphoma, the most common subtype of non-Hodgkin lymphoma, presents major therapeutic challenges due to molecular heterogeneity and high relapse rates. While autologous hematopoietic stem cell transplantation (ASCT) has been a cornerstone for relapsed/refractory (R/R) B-cell lymphoma, its efficacy is often compromised by minimal residual disease (MRD) persistence and an immunosuppressive tumor microenvironment. Chimeric antigen receptor (CAR)-T cell therapy has transformed treatment paradigms but faces limited long-term durability due to antigen escape and T-cell exhaustion. The integration of ASCT with CAR-T therapy may offer a complementary approach to address these limitations, leveraging ASCT-induced immune reconstitution to enhance CAR-T-cell persistence and reprogram the tumor milieu. Emerging clinical evidence supports this approach, indicating improved disease control and progression-free survival. Although preliminary clinical outcomes are encouraging, unresolved challenges persist, particularly in terms of cumulative toxicity, optimal therapeutic sequencing, CAR-T-cell longevity, and financial feasibility associated with these advanced therapies. This review provides a comprehensive overview of mechanistic synergies between ASCT and CAR-T therapy, critically evaluates emerging clinical evidence on treatment sequencing, and explores innovative strategies to increase safety, efficacy, and accessibility.

Organoids are three-dimensional (3D) cell cultures derived from human pluripotent stem cells or adult stem cells that recapitulate the cellular heterogeneity, structure, and function of human organs. These microstructures are invaluable for biomedical research due to their ability to closely mimic the complexity of native tissues while retaining human genetic material. This fidelity to native organ systems positions organoids as a powerful tool for advancing our understanding of human biology and for enhancing preclinical drug testing. Recent advancements have led to the successful development of a variety of organoid types, reflecting a broad range of human organs and tissues. This progress has expanded their application across several domains, including regenerative medicine, where organoids offer potential for tissue replacement and repair; disease modeling, which allows for the study of disease mechanisms and progression in a controlled environment; drug discovery and evaluation, where organoids provide a more accurate platform for testing drug efficacy and safety; and microecological research, where they contribute to understanding the interactions between microbes and host tissues. This review provides a comprehensive overview of the historical development of organoid technology, highlights the key achievements and ongoing challenges in the field, and discusses the current and emerging applications of organoids in both laboratory research and clinical practice.

Considering the limitations of intraportal transplantation (Tx), we sought to establish an alternative approach for it-transplanting islets onto the liver surface (LS) by optimizing adipose tissue-derived stem cell (ADSC) co-Tx procedures with a gelatin hydrogel nonwoven fabric (GHNF). In the in vivo study, we examined the use of the GHNF, the effectiveness of islet covering materials, and preferred procedures for ADSC co-Tx using a syngeneic rat model. Immunohistochemical staining was performed to evaluate the extracellular matrix (ECM) expression and angiogenesis. In the in vitro study, we analyzed the culture supernatants to identify crucial factors secreted from ADSCs in different ADSC co-Tx procedures. It was shown that the GHNF should be used to cover the islets but not to embed internally (encapsulate) them. Utilization of the GHNF in LS Tx resulted in significantly better glucose changes (P = 0.0002) and cure rate of diabetic recipients (P = 0.0003) than the use of a common adhesion barrier. Although neovascularization was comparable among groups, ECM reconstitution tended to be higher when the GHNF was used. ADSC co-Tx further enhanced ECM reconstitution only when ADSCs were cultured in the GHNF before islet Tx. Leptin, vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and several chemokines were identified as candidate factors for enhancing ECM reconstitution (P < 0.001). The inhibition assay using antagonist suggested that leptin might be at least in part responsible for the difference in transplant efficiency in distinct ADSC co-Tx methods. This study showed that the GHNF effectively improved the outcomes of LS islet Tx, mainly due to ECM reconstitution around the islets. Furthermore, we established a novel method of LS islet Tx by combining a GHNF with ADSCs, which is equally effective as intraportal Tx.

Long noncoding RNAs (lncRNAs) are crucial in porcine preimplantation embryonic development, yet their regulatory role during zygote genome activation (ZGA) is poorly understood. We analyzed transcriptome data from porcine fetal fibroblasts (PEF), induced pluripotent stem cells (iPS), and preimplantation embryos, identifying ZGA-specific lncRNAs like XLOC-040580, and further predicted its potentially interacting genes TPRA1 and BCL2L1 via co-expression network. XLOC-040580 was knocked down by siRNA microinjection and the expression of ZGA-related genes was detected by qRT-PCR. After microinjecting siRNA targeting TPRA1 and BCL2L1 at the one-cell stage, we counted the blastocyst development rate. The blastocyst development rate was consistent with the results from si-XLOC-040580 after si-TPRA1. Through dual-luciferase reporter assays, we found that XLOC-040580 was a downstream target of TPRA1. To further elucidate the mechanism of XLOC-040580, Single-cell mRNA sequencing after XLOC-040580 knockdown revealed its regulatory network involved in embryonic developmental defects. Transcriptome analysis revealed that XLOC-040580 was specifically expressed during zygote activation. Knockdown of XLOC-040580 decreased the blastocyst development rate and reduced both the total blastocyst cell number and TE cell number. TPRA1 and BCL2L1 were specifically co-expressed with XLOC-040580 during ZGA stage, and TPRA1 could interact with the promoter region of XLOC-040580 and regulate its expression. Knockdown of TPRA1 or XLOC-040580 blocked porcine embryonic development by affecting the expression of ZGA-related genes. We found and validated that lncRNA XLOC-040580 played a key role in the ZGA process, which was regulated by TPRA1. These results implied that the functional axis of TPRA1-XLOC-040580-downstream genes involved in ZGA-related functions also coordinated early embryonic development in porcine.