Gastric cancer remains a leading cause of cancer-related mortality, with limited treatment options and a poor prognosis. Mesenchymal stem cells (MSCs) possess inherent tumor-homing ability but exhibit modest efficacy against solid tumors. Enhancing both tumor targeting and immune activation through genetic modification may improve the therapeutic potential of these approaches. To evaluate the anti-cancer efficacy of human umbilical cord–derived MSCs (HUC-MSCs) genetically modified to co-express C-X-C chemokine receptor type 4 (CXCR4) and TNFSF14 (LIGHT) against gastric cancer in vitro and in vivo. HUC-MSCs were transduced via lentiviral vectors to generate dual-gene–engineered Tg-HUC-MSCs. In vitro, their effects on HGC-27 gastric cancer cells were assessed using the scratch wound migration assay and Annexin V/PI apoptosis analysis. In vivo, BALB/c nude mice bearing subcutaneous HGC-27 xenografts were treated with Tg-HUC-MSCs, unmodified HUC-MSCs, or a control. Tumor growth was monitored by caliper measurements. Excised tumors were analyzed histologically with hematoxylin–eosin staining and by Ki-67 immunohistochemistry. Tg-HUC-MSCs significantly inhibited HGC-27 cell migration and induced higher apoptosis rates than unmodified HUC-MSCs in vitro (p < 0.001). In vivo, Tg-HUC-MSCs markedly reduced tumor volume, increased necrotic areas, and decreased Ki-67 proliferation indices compared to controls (p < 0.001). Elevated CXCR4 and LIGHT expression correlated with improved tumor targeting and enhanced immune-mediated tumor cell killing. Dual-gene–engineered HUC-MSCs (CXCR4 + TNFSF14) demonstrated superior tumor-homing and anti-cancer activity against gastric cancer cells in vitro and in vivo, representing a promising cell-based therapeutic strategy for solid tumors.

Cellular morphology, a critical manifestation of biological characteristics, is linked to functions. In traditional cell detection, invasive labeling and detection methods not only compromise cellular viability but also entail labor-intensive workflows. Here we presented a non-invasive artificial intelligence framework that integrated deep learning (DL) and machine learning (ML) to predict the immunomodulatory capacity of mesenchymal stem cells (MSCs) through morphological profiling. The improved PreAct-ResNet50 encoder-decoder architecture was used to achieve high-accuracy instance segmentation of cells and nuclei, enabling quantification of morphological features. A LightGBM-based predictive model was subsequently employed to predict MSCs immunomodulatory biomarkers through morphological features. This dual-model system demonstrated satisfactory cell segmentation and biological characteristics prediction capabilities through performance testing. Our method provided an efficient, non- invasive tool for real-time MSCs potency assessment, which could enhance quality controls in cell therapy manufacturing.

BackgroundThis study aims to investigate the application of visual information processing mechanisms in the segmentation of stem cell (SC) images. The cognitive principles underlying visual information processing were analyzed, and the limitations of conventional segmentation methods were evaluated using phase-contrast microscopy images of stem cells.MethodsAn optimized segmentation method incorporating halo correction was developed to address the limitations of traditional approaches. The performance of the proposed method was experimentally validated and compared with existing techniques.ResultsThe proposed method achieved segmentation accuracy, recall, precision, and F1-score values of 96.5%, 94.9%, 91.4%, and 93.9%, respectively, outperforming existing approaches. Additionally, the confluency error on the Human Mesenchymal Stem Cells dataset and the C2C12 dataset was 0.07 and 0.05, respectively, indicating superior performance compared to equivalent methods.ConclusionThe findings demonstrate that the proposed method offers enhanced efficacy for stem cell image segmentation tasks.

The development of robust and scalable culture systems is essential for the clinical-scale production of human umbilical cord (UC)-derived mesenchymal stem/stromal cells (MSCs) (UC-MSCs). While various basal and serum-free media are commercially available, systematic comparisons of their efficacy in supporting the expansion and functional properties of UC-MSCs remain limited. In this study, we conducted a comprehensive evaluation of multiple culture systems, including basal media (α-MEM, DMEM, and DMEM/F12) supplemented with human platelet lysate (HPL), and commercial serum-free media (Corning MSC Xeno-Free SFM, NutriStem XF Medium, Prime-XV MSC Expansion XSFM), for their ability to sustain UC-MSCs proliferation, maintain phenotypic properties, and support functional potency. The results demonstrated that all basal media supported cell growth, with α-MEM (Gibco) and DMEM/F12 showing superior performance over DMEM. Among serum-free formulations, Prime-XV with 2% HPL yielded the highest primary culture output and the shortest population doubling (PD) time (PDT) during passaging. Notably, cells expanded in commercial serum-free media exhibited reduced diameter and higher uniformity. Functional analyses revealed that NutriStem XF Medium supplemented with 2% HPL elicited the strongest immunomodulatory effects in mixed lymphocyte reactions (MLRs). Furthermore, all media maintained trilineage differentiation capacity and satisfied International Society for Cellular Therapy (ISCT) phenotypic criteria. Critically, no tumorigenic potential was detected in vitro or in vivo. Large-scale manufacturing using the selected medium (NutriStem XF + 2% HPL) confirmed consistent expansion kinetics, high viability, stable marker expression, and functional potency across seven production batches. This study provides a rigorous and clinically relevant framework for selecting culture media that ensure both scalability and functional integrity of UC-MSCs, highlighting the promise of serum-free systems for therapeutic manufacturing.

BACKGROUND:Osteoarthritis (OA) is a common cause of disability around the world, but the pathophysiology is still poorly understood. The study sought to investigate the effects of umbilical cord mesenchymal stem cells (UC-MSCs) injection in OA, with a focus on cartilage degradation. METHODS:The serum matrix metalloproteinase (MMPs) and a disintegrin and metalloprotease domains with thrombospondins motifs (ADAMTS) levels in OA patients were examined using ELISA. The levels of MMPs and ADAMTS in peripheral blood mononuclear cells (PBMCs) following co-cultured with UC-MSC were determined by ELISA. The anterior cruciate ligament transection (ACLT) surgery was performed on the knee joints of a rat OA model, followed by intra-articular injection of UC-MSCs at 4 weeks and 8 weeks after surgery, and the changes of the severity of OA, tibiofemoral cartilage, and subchondral bone were observed by Safranin-O staining, hematoxylin and eosin (H&E) staining, and micro-CT imaging. RESULTS:OA patients showed the higher protein levels of MMP2, 9, 13, ADAMTS4 and 5 than healthy controls. Furthermore, in vitro incubation of PBMC derived from OA patients with UC-MSCs down-regulated the protein expression of these proteases. Positive correlations were observed between serum MMP9 levels and high-sensitivity C-reactive protein (hsCRP) in OA patients, while a negative correlation between serum MMP13 and Alkaline phosphatase (ALKP) was observed in these patients. There was a negative correlation between OA patients' serum ADAMTS 4 and Lipoprotein (a)(Lp(a)). Western blotting and immunohistostaining were applied to assess the protein levels of matrix metalloproteinases and ADAMTSs in rat knee joints and these protein levels were significantly decreased in the UC-MSC intra-articular injection group. Micro-CT results showed in the OA rat model human UC-MSCs treatment alleviated the destruction of the tibial subchondral bone. CONCLUSION:UC-MSCs decreased the level of several matrix proteases, slowing the progression of OA formation in rats. Our findings suggested that matrix metalloproteinases and ADAMTSs play a role in OA progression, UC-MSC exerted beneficial therapeutic effects on OA rat model by reducing the levels of these matrix metalloproteinases and ADAMTSs.

Abstract Background Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) hold great therapeutic potential in regenerative medicine. Therefore, it is crucial to establish a Good Manufacturing Practice (GMP)-compliant methodology for the isolation and culture of WJ-MSCs. Through comprehensive research, encompassing laboratory-scale experiments to pilot-scale studies, we aimed to develop standardized protocols ensuring the high yield and quality of WJ-MSCs manufacturing. Methods Firstly, optimization of parameters for the enzymatic digestion method used to isolate WJ-MSCs was conducted. These parameters included enzyme concentrations, digestion times, seeding densities, and culture media. Additionally, a comparative analysis between the explant method and the enzymatic digestion method was performed. Subsequently, the consecutive passaging of WJ-MSCs, specifically up to passage 9, was evaluated using the optimized method. Finally, manufacturing processes were developed and scaled up, starting from laboratory-scale flask-based production and progressing to pilot-scale cell factory-based production. Furthermore, a stability study was carried out to assess the storage and use of drug products (DPs). Results The optimal parameters for the enzymatic digestion method were a concentration of 0.4 PZ U/mL Collagenase NB6 and a digestion time of 3 h, resulting in a higher yield of P0 WJ-MSCs. In addition, a positive correlation between the weight of umbilical cord tissue and the quantities of P0 WJ-MSCs has been observed. Evaluation of different concentrations of human platelet lysate revealed that 2% and 5% concentrations resulted in similar levels of cell expansion. Comparative analysis revealed that the enzymatic digestion method exhibited faster outgrowth of WJ-MSCs compared to the explant method during the initial passage. Passages 2 to 5 exhibited higher viability and proliferation ability throughout consecutive passaging. Moreover, scalable manufacturing processes from the laboratory scale to the pilot scale were successfully developed, ensuring the production of high-quality WJ-MSCs. Multiple freeze-thaw cycles of the DPs led to reduced cell viability and viable cell concentration. Subsequent thawing and dilution of the DPs resulted in a significant decrease in both metrics, especially when stored at 20–27 °C. Conclusion This study offers valuable insights into optimizing the isolation and culture of WJ-MSCs. Our scalable manufacturing processes facilitate the large-scale production of high-quality WJ-MSCs. These findings contribute to the advancement of WJ-MSCs-based therapies in regenerative medicine.

Cell counting is a common and fundamental cell measurement technique that plays a crucial role in the development and quality control of cell therapy products. However, accurate and reliable cell counting can be challenging owing to the complexity of cell preparations, diverse counting purposes, and various counting methods. This review summarizes the challenges encountered in cell counting for cell therapy products and provides strategies to improve the cell counting accuracy, thereby guiding the counting process and ensuring the quality of cell therapy products.

目的 验证基于吖啶橙(AO)/碘化丙啶(PI)荧光染色原理的自动细胞分析仪检测细胞因子诱导的杀伤细胞(CIK)数量及活率的可行性,并对CIK细胞培养全过程及冻存复苏后至使用前过程进行检测,建立相应的数量及活率标准.方法 采集健康供者的外周血分离、培养CIK细胞,取培养过程中的细胞使用AO/PI荧光染色法、应用自动细胞分析仪检测细胞数量及活率,对结果的专属性、准确度、精密度、数量线性及范围、活率线性及范围进行验证.应用建立的方法对外周血分离后的外周血单个核细胞(PBMC)、培养过程中、冻存前及复苏后的CIK细胞数量及活率进行全过程检测.结果 专属性、准确度、精密度、数量线性及范围、活率线性及范围验证均符合预期要求.CIK细胞培养全过程活率均不低于80％,样本平均倍增时间为(42.26±1.17)h.冻存前的CIK细胞在加入含5％DMSO的冷冻保护剂后90min内活率稳定,复苏后未经稀释或洗涤处理120min内活率稳定,复苏后经洗涤去除DMSO处理后12 h内活率稳定.结论 基于AO/PI荧光染色原理的自动细胞分析仪可以用于检测CIK细胞的数量及活率,其结果可以用于细胞放行评价及稳定性研究.

Coronavirus disease 2019 (COVID-19) treatments are still urgently needed for critically and severely ill patients. Human umbilical cord-mesenchymal stem cells (hUC-MSCs) infusion has therapeutic benefits in COVID-19 patients; however, uncertain therapeutic efficacy has been reported in severe patients. In this study, we selected an appropriate cytokine, IL-18, based on the special cytokine expression profile in severe pneumonia of mice induced by H1N1virus to prime hUC-MSCs in vitro and improve the therapeutic effect of hUC-MSCs in vivo. In vitro, we demonstrated that IL-18-primed hUC-MSCs (IL18-hUCMSC) have higher proliferative ability than non-primed hUC-MSCs (hUCMSCcon). In addition, VCAM-1, MMP-1, TGF-β1, and some chemokines (CCL2 and CXCL12 cytokines) are more highly expressed in IL18-hUCMSCs. We found that IL18-hUCMSC significantly enhanced the immunosuppressive effect on CD3 + T-cells. In vivo, we demonstrated that IL18-hUCMSC infusion could reduce the body weight loss caused by a viral infection and significantly improve the survival rate. Of note, IL18-hUCMSC can also significantly attenuate certain clinical symptoms, including reduced activity, ruffled fur, hunched backs, and lung injuries. Pathologically, IL18-hUCMSC transplantation significantly enhanced the inhibition of inflammation, viral load, fibrosis, and cell apoptosis in acute lung injuries. Notably, IL18-hUCMSC treatment has a superior inhibitory effect on T-cell exudation and proinflammatory cytokine secretion in bronchoalveolar lavage fluid (BALF). Altogether, IL-18 is a promising cytokine that can prime hUC-MSCs to improve the efficacy of precision therapy against viral-induced pneumonia, such as COVID-19.

【视频简介】 当今世界下，国际细胞相关研究正如火如荼的开展，目前细胞研究方向除了细胞治疗、组织器官移植修复、基因治疗之外，还向着药物研发、毒性评估工具、发育生物学模型等领域转变。随着细胞基本原理和相关技术的成熟和更新，以及监管政策的不断转暖，各国已纷纷加快细胞的临床研究，并将其列入国家科技战略必争领域。本视频介绍了如今国际细胞治疗产业发展的现状及面临的机遇与挑战，从A面来看：产业链条相对完善，投融资活动活跃，资本市场看好，不断有新产品获批上市；从B面来讲这种发展是一种非理性繁荣，在这背后更是面临"未被满足"的临床需求。基于国际发展的现状，刘沐芸博士对国内细胞治疗产业"未来该何去何从"提出了几点看法与建议，应当将产业发展的A、B面进行协调统一，通过数字化、自动化与智能化实现细胞产业发展的提质增效，提供终极解决方案。

Objective: Human umbilical cord mesenchymal stem cells (hUC-MSCs) have shown very attractive potential in clinical applications for the treatment of various diseases. However, the data about the reproductive and developmental toxicity of hUC-MSCs remains insufficient. Thus, we assessed the potential effects of intravenous injection of hUC-MSCs on reproduction and development in Sprague-Dawley rats.Methods: In the fertility and early embryonic development study, hUC-MSCs were administered at dose levels of 0, 6.0 × 106, 8.5 × 106, and 1.2 × 107/kg to male and female rats during the pre-mating, mating and gestation period. In the embryo-fetal development study, the pregnant female rats received 0, 6.0 × 106, 1.2 × 107, and 2.4 × 107/kg of hUC-MSCs from gestation days (GD) 6–15. Assessments made included mortality, clinical observations, body weight, food consumption, fertility parameters of male and female, litter, and fetus parameters, etc.Results: No hUC-MSCs-related toxicity was observed on the fertility of male and female rats, and no teratogenic effect on fetuses. hUC-MSCs at 1.2 × 107/kg caused a mildly decrease in body weight gain of male rats, transient listlessness, tachypnea, and hematuria symptoms in pregnant female rats. Death was observed in part of the pregnant females at a dose of 2.4 × 107/kg, which could be due to pulmonary embolism.Conclusion: Based on the results of the studies, the no-observed-adverse-effect levels (NOAELs) are 8.5 × 106/kg for fertility and early embryonic development, 1.2 × 107/kg for maternal toxicity and 2.4 × 107/kg for embryo-fetal development in rats intravenous injected with hUC-MSCs, which are equivalent to 8.5-fold, 12-fold, and 24-fold respectively of its clinical dosage in humans. These findings may provide a rational basis for human health risk assessment of hUC-MSCs.

Background： Pulmonary fibrosis (PF) is a growing clinical problem with limited therapeutic options. Human umbilical cord mesenchymal stromal cell (hucMSC) therapy is being investigated in clinical trials for the treatment of PF patients. However, little is known about the underlying molecular and cellular mechanisms of hucMSC therapy on PF. In this study, the molecular and cellular behavior of hucMSC was investigated in a bleomycin induced mouse PF model. Methods: The effect of hucMSC on mouse lung regeneration was determined by detecting Ki67 expression and Edu incorporation in alveolar type 2 (AT2) and lung fibroblast cells. The hucMSC was transfected to express the membrane localized GFP before transplant into the mouse lung. The cellular behavior of hucMSC in mouse lung was tracked by GFP staining. Single cell RNA sequencing was performed to investigate the molecular mechanism of hucMSC on PF. Results: hucMSCs could alleviate collagen accumulation in lung and decrease the mortality of mouse induced by bleomycin. hucMSCs transplantation promoted AT2 cell proliferation and inhibited lung fibroblast cell proliferation. Mouse lung macrophage phenotype was changed after interacting with hucMSCs. By using single-cell RNA sequencing, a subcluster of interferon-sensitive macrophages were identified after hucMSC infusion. These macrophages elevate the secretion of CXCL9 and CXCL10 following hucMSC infusion and recruit more Treg cells to the injured lung. Conclusions: Our study establishes a link between hucMSCs, macrophage proliferation, and PF with potential applications in PF therapeutics. It provides new insights into how hucMSCs interact with macrophage during the repair process of bleomycin-induced PF and play its immunoregulation function.

BACKGROUND:Pulmonary fibrosis (PF) is a growing clinical problem with limited therapeutic options. Human umbilical cord mesenchymal stromal cell (hucMSC) therapy is being investigated in clinical trials for the treatment of PF patients. However, little is known about the underlying molecular and cellular mechanisms of hucMSC therapy on PF. In this study, the molecular and cellular behavior of hucMSC was investigated in a bleomycin-induced mouse PF model.METHODS:The effect of hucMSCs on mouse lung regeneration was determined by detecting Ki67 expression and EdU incorporation in alveolar type 2 (AT2) and lung fibroblast cells. hucMSCs were transfected to express the membrane localized GFP before transplant into the mouse lung. The cellular behavior of hucMSCs in mouse lung was tracked by GFP staining. Single cell RNA sequencing was performed to investigate the effects of hucMSCs on gene expression profiles of macrophages after bleomycin treatment.RESULTS:hucMSCs could alleviate collagen accumulation in lung and decrease the mortality of mouse induced by bleomycin. hucMSC transplantation promoted AT2 cell proliferation and inhibited lung fibroblast cell proliferation. By using single cell RNA sequencing, a subcluster of interferon-sensitive macrophages (IFNSMs) were identified after hucMSC infusion. These IFNSMs elevate the secretion of CXCL9 and CXCL10 following hucMSC infusion and recruit more Treg cells to the injured lung.CONCLUSIONS:Our study establishes a link between hucMSCs, macrophage, Treg, and PF. It provides new insights into how hucMSCs interact with macrophage during the repair process of bleomycin-induced PF and play its immunoregulation function.

Mesenchymal stromal cells (MSCs) show potential for treating preclinical models of newborn bronchopulmonary dysplasia (BPD), but studies of their therapeutic effectiveness have had mixed results, in part due to the use of different media supplements for MSCs expansion in vitro. The current study sought to identify an optimal culture supplement of umbilical cord-derived MSCs (UC-MSCs) for BPD therapy. In this study, we found that UC-MSCs cultured with human platelet lysate (hPL-UCMSCs) were maintained a small size from Passage 1 (P1) to P10, while UC-MSCs cultured with fetal bovine serum (FBS-UCMSCs) became wide and flat. Furthermore, hPL was associated with lower levels of senescence in UC-MSCs during in vitro expansion compared with FBS, as indicated by the results of β-galactosidase staining and measures of senescence-related genes (CDKN2A, CDKN1A, and mTOR). In addition, hPL enhanced the proliferation and cell viability of the UC-MSCs and reduced their doubling time in vitro. Compared with FBS-UCMSCs, hPL-UCMSCs have a greater potential to differentiate into osteocytes and chondrocytes. Moreover, using hPL resulted in greater expression of Nestin and specific paracrine factors (VEGF, TGF-β1, FGF2, IL-8, and IL-6) in UC-MSCs compared to using FBS. Critically, we also found that hPL-UCMSCs are more effective than FBS-UCMSCs for the treatment of BPD in a rat model, with hPL leading to improvements in survival rate, lung architecture and fibrosis, and lung capillary density. Finally, qPCR of rat lung mRNA demonstrated that hPL-UCMSCs had lower expression levels of inflammatory factors (TNF-α and IL-1β) and a key chemokine (MCP-1) at postnatal day 10, and there was significant reduction of CD68+ macrophages in lung tissue after hPL-UCMSCs transplantation. Altogether, our findings suggest that hPL is an optimal culture supplement for UC-MSCs expansion in vitro, and that hPL-UCMSCs promote lung repair in rat BPD disease.

Objective:To explore the immunosuppressive ability of human umbilical cord mesenchymal stem cells (hUCMSCs) on T and B lymphocytes before and after cryopreservation and provide future for using the clinical application of hUCMSCs.Methods:Human peripheral blood mononuclear cells were isolated from healthy human peripheral blood. Anti-CD3 and anti-CD8 monoclonal antibodies were used to activate T lymphocytes in vitro. Selected B lymphocytes were stimulated by ODN2395, human CD40 ligand, goat anti-human IgM antibodies and interleukin 2 in vitro. The cells were co-cultured with activated T and B lymphocytes, divided into a single stimulation group and hUCMSCs co-culture group (before and after cryopreservation) . The immunosuppressive ability of hUCMSCs on activated T and B lymphocytes were analyzed. In addition, the differences immunosuppressive ability of hUCMSCs before and after cryopreservation were also studied. The differences between groups were compared by t-test, and the differences among groups were compared by repeated measures design analysis of variance.Results:Compared with hUCMSCs before cypropreservation, the proliferation of CD3+T cells (37.60﹪±0.54﹪vs 39.40﹪±1.57﹪) , CD4+T cells (36.87﹪±0.54﹪vs 38.63﹪±1.39﹪) and CD8+T cells (40.37﹪±1.14﹪vs 42.47﹪±1.90﹪) (P > 0.05) were not significantly inhibited after cryopreservation. The apoptosis of CD3+T cells (18.07﹪±0.66﹪vs 16.77﹪±1.15﹪) , CD4+T cells (26.47﹪±1.13﹪vs 24.60﹪±1.47﹪) and CD8+T cells (3.52﹪±0.22﹪vs 2.72﹪±0.06﹪) had no significant difference (P > 0.05) . The proportion of regulatory cells (Treg) subsets (5.51﹪±0.71﹪vs 6.87﹪± 0.27﹪) had also no significant difference (P > 0.05) . hUCMSCs did not reduce the proportion of helper T cell (Th) subsets, especially the proportion of Th1 subsets (0.47﹪± 0.09﹪vs 0.33﹪± 0.04﹪) and Th17 subsets (0.21﹪± 0.04﹪vs 0.22﹪± 0.03﹪) (P > 0.05) . For B lymphocytes, hUCMSCs reduced the proportion of Th2 cell subsets (0.73﹪±0.07﹪vs 0.49﹪±0.06﹪) and IgM antibody secretion [ (739.70±28.39) ng/mL vs (560.21±3.81) ng/mL] after cryopreservation (P < 0.05) . There was no significant difference in proliferation and apoptosis of B lymphocytes, and reduced secretion of IgA and IgG antibodies before and after hUCMSCs cytopreservation (P > 0.05) .Conclusion:After cryopreservation, hUCMSCs were more powerful than those before cryopreservation in reducing the proportion of Th2 cell subsets and the secretion of IgM antibodies. However, the immunosuppressive abilities was same. Both hUCMSCs before and after cryopreservation have high clinical application value.

目的 比较临床检测策略(CDS)与血筛检测策略(BSS)在综合细胞库供体病原体检测中的差异,进行研究分析.方法 对深圳综合细胞库2016年2月至2017年7月共计7981例细胞供者的血浆标本进行病原体检测,病原体类型包括乙型肝炎病毒(HBV)、丙型肝炎病毒(HCV)、人类免疫缺陷病毒(HIV1/2)与梅毒螺旋体(TP)四类.实验采用CDS和BSS两种检测策略进行.分析两者在病原体总体检测结果及各单项检测结果的一致性,比较两种检测策略的差异性.同时分析BSS中血清学ELISA与核酸检测(NAT)方法对HBV、HCV、HIV检测结果的一致性,比较两者的差异性.结果 BSS与CDS两种检测策略的一致性一般(K=0.563),差异有统计学意义(P<0.05).两种检测策略中,HBV的一致性一般(K=0.528),HCV的一致性很差(K=0.105),HIV1/2的一致性很差(K=0);比较BSS中血清学检测方法与核酸检测方法,两种检测方法的一致性较好(K=0.651),但两种方法差异有统计学意义(P<0.05),其中HBV的血清学检测与核酸检测两种方法的一致性非常好(K=0.833),而HCV与HIV血清学检测与核酸检测两种方法的一致性较差(K<0.4).结论 BSS在细胞供者病原体检测种类上更全面,检测灵敏度更高;在细胞库病原体污染风险防控中应同时重视两种检测方法试剂的选择,整体上降低任何单一检测方法不准确导致的病原体漏检风险,避免病原体交叉污染,保证细胞库储存样本安全.

目的 从细胞学方面探讨治疗新冠肺炎患者临床使用的人脐带间充质干细胞(hUC-MSCs)在体外连续制备过程中其细胞因子和免疫调节蛋白的基础分泌水平,以及探究IFN-γ刺激hUC-MSCs后对细胞因子和免疫调节蛋白的影响.方法 首先,利用ELISA等方法检测在连续传代扩增hUC-MSCs中多种细胞因子(IL-6、IL-1β、TGF-β、HGF和PGE-2)和免疫调节蛋白(PD-L1和IDO1)的基础表达水平和稳定性;其次,探究hUC-MSCs受到炎症因子IFN-γ刺激后因子分泌和蛋白表达的变化情况.结果 hUC-MSCs在连续传代中其9种MSC相关表面标志物没有明显改变;PO代hUC-MSCs分泌高水平的HGF和TGF-β,但会随着传代增加而分泌水平显著下降;而IL-6和PGE-2则会随着传代增加而分泌上升;IL-1β总体基础表达水平不高;同时,WB和流式结果显示,免疫抑制相关的蛋白IDO1和PD-L1在正常情况下几乎不表达.而当hUC-MSCs受到炎症因子IFN-γ刺激后,细胞因子分泌普遍没有明显上调,其中HGF和PGE-2刺激后还出现了显著下降的趋势;但是免疫抑制相关蛋白IDO1和PD-L1的表达却显著上升,具有统计学意义.结论 hUC-MSCs在连续传代过程中不同细胞因子的表达水平变化具有一定的差别.在受到炎症因子IFN-γ的激活后,可能通过提高IDO1和PD-L1的蛋白表达来发挥免疫抑制的作用.此结果为hUC-MSCs治疗新冠肺炎临床疗效提供理论依据.

新冠肺炎疫情的发生,可能让平日胸有成竹的我们突然感到手足无措和人生无常.病毒很小,却给我们带来了极大的不安全感,在某种程度上也诱发我们思考,面对未来可能的突发情况,我们应防患于未然还是水来土掩? 新冠肺炎的爆发让我们回想起,应对2003年SARS流行和2009年H1N1大流行时的若干建议,要改善收入不平等、公共卫生地区差异,研发疫苗和治疗药物,以及要提升我们对威胁来临前的及时洞察能力和网络建设等.从时间来看,从SARS到H1N1,再到这次COVID-19,可能并不是我们遗忘了威胁本身,而是中间间隔的时间有点儿长,导致我们会形成一种应答式的应对方式(Crisis-Response):威胁来袭时,我们会过度反应并全力应对,当威胁解除后,我们就会逐渐淡忘,甚至会忘记威胁曾经来过,然后重复老路.如何打破这个“惊恐——忽略”的应对循环呢?

Peripheral blood mononuclear cells are widely used as source material for anticancer immunotherapies. The conventional cryopreservation method for peripheral blood mononuclear cells is time-consuming and expansive, which involves controlled rate freezing followed by storage in liquid nitrogen. Instead, the convenient uncontrolled rate freezing cryopreservation method had been reported successfully in peripheral blood hematopoietic stem cells and peripheral blood progenitor cells. Therefore, we hypothesized that uncontrolled rate freezing cooling method maybe also applied to peripheral blood mononuclear cells cryopreservation. In this study, we evaluated the performance of uncontrolled rate freezing and controlled rate freezing cooling methods through cell recovery rate, viability, differentiation potential into cytokine-induced killer cells and the cellular properties of the cultured cytokine-induced killer cells. The results showed similar post-thaw viability and recovery rate in both controlled rate freezing and uncontrolled rate freezing cryopreserved peripheral blood mononuclear cells. Importantly, the uncontrolled rate freezing cryopreserved peripheral blood mononuclear cells exhibited higher growth ratio and earlier cell clustering during ex-vivo cytokine-induced killer cell culture than the controlled rate freezing ones. These two groups of expanded cytokine-induced killer cells also exhibited similar effector cell subset ratio and tumoricidal activity. In general, the performance of cryopreserved peripheral blood mononuclear cells using uncontrolled rate freezing cooling method, with the commercial cryoprotective agent CellBanker 2, was equal or better than the controlled rate freezing method. Our study implied that the combined use of cryoprotective agent CellBanker 2 and uncontrolled rate freezing could be a convenient cryopreservation method for peripheral blood mononuclear cells.

Mesenchymal stem cells (MSC) are a popular candidate in cellular therapy for many diseases. MSCs are well known by their feature of self-renewal and their differentiation potential. NESTIN is a cytoskeletal protein expressed in MSC that functions directly in cell proliferation and differentiation. Here, we demonstrated that adding UltraGRO, a medium supplement, could maintain and partially recover the expression of NESTIN in human umbilical cord derived MSCs (UC-MSCs). Furthermore, the UC-MSCs cultured with UltraGRO showed a better immunomodulation ability in a colitis mouse model compared with those cultured in other types of media. This indicates that the use of novel culture medium benefits the maintenance of NESTIN expression and NESTIN may be one of the vital factors that regulates the performance of MSCs.