Cell Transplantation最新文献

Cell transplantation is a promising approach for addressing neurodegenerative conditions. In this study, we developed a robot-assisted stereotactic microinjection system for transplanting cells. We evaluated the factors that affect cellular graft viability and other properties, including the gauge of the syringe needle and the injection rate. We systematically compared the synchronous withdrawal injection (SWI) and fixed-point injection (FPI) procedures in agarose and rat brain models. In vitro assessments revealed superior dye dispersion with SWI compared to FPI, and in vivo analyses confirmed that SWI reduced the tissue injury and improved cell distribution in the striatum. We applied this robot-assisted technique to evaluate the accuracy and safety of cell transplantation in canine models. Overall, this strategy enhances the accuracy and safety of graft delivery, potentially improving outcomes and advancing therapeutic strategies for the clinical treatment of neurodegenerative disorders.

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.

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.

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.

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, which are three-dimensional cultures derived from pluripotent or adult stem cells, meticulously mimic human organ architecture and function, revolutionizing biomedical research. Patient-derived organoids have emerged as powerful tools in disease modeling, particularly in cancer research. For instance, in the case of colorectal cancer, organoids are developed from tumor tissues of patients, allowing for drug sensitivity tests that can inform personalized treatment plans, as seen in the case of a patient responding well to specific chemotherapy drugs. In addition, organoids have been used to study regenerative mechanisms, such as the repair of intestinal stem cells post-radiation, showcasing their versatility in biomedical research. In drug development, they facilitate screening for efficacy and toxicity, with applications in testing poly-ADP ribose polymerase (PARP) inhibitors and cosmetic ingredients, while aligning with ethical imperatives as the U.S. Food and Drug Administration (FDA) plans to phase out animal testing for certain drugs by 2025. In addition, organoids show promise in regenerative medicine, such as endometrial and retinal regeneration, and bone tissue engineering. Despite challenges such as variable culture conditions, limited vascularization, and high costs, standardizing protocols and integrating microenvironmental factors will enhance their clinical utility, driving a shift toward human-centric therapeutic advancements.

Cesarean section (CS) scar tissue fibrosis and decreased muscle density increase the risk of uterine rupture and placental implantation in subsequent pregnancies. Given the potent anti-fibrotic and regenerative properties of human amniotic mesenchymal stem cells (hAMSCs), this study investigated their therapeutic potential in repairing uterine scars in a rat model. A full-thickness uterine wall excision model was established to mimic CS scarring. hAMSCs were transplanted at the edge of the incision. Myometrial thickness and collagen deposition within the scar were assessed histologically using hematoxylin and eosin (H&E) and Masson's staining. Immunohistochemistry evaluated the expression of MMP8, TGF-β1, VEGFA, and α-SMA within the scar region. mRNA transcriptome sequencing and quantitative real-time polymerase chain reaction (qRT-PCR) were employed to explore possible mechanisms. A total of 110 rats were used in the study, 30 in the sham group, 40 in the phosphate-buffered saline (PBS) control group and 40 in the hAMSCs treatment group. Compared with the PBS group, the hAMSCs group exhibited a 35% reduction in collagen fiber area and a 28% increase in smooth muscle cell density (P < 0.05). The MMP8, VEGFA, and α-SAM expressions in the uterine scar area of rats increased, whereas the TGF-β1 expression decreased (P < 0.05). Transcriptome sequencing and real-time fluorescence quantitative PCR results showed that the expression levels of Wnt4, Fzd5, Wnt5a, and PPARD genes were lower in the uterine scar region of rats in the hAMSCs group compared with those in the PBS group. Transplantation of hAMSCs inhibits scar formation in uterine wounds and promotes regeneration of smooth muscle tissue and neovascularization, which in turn promotes uterine wound repair. This effect may be related to the activation of Wnt pathway and the inhibition of PPARD gene expression.