albumin

JUNCTION CITY, Kan.--(BUSINESS WIRE)--InVitria, a leading developer of recombinant, chemically defined components for biomanufacturing, has announced the launch of Optibumin® 25, the first and only recombinant human serum albumin (rHSA) available as a 25% solution. Designed for closed-system workflows, Optibumin 25 is a high-purity alternative to plasma-derived albumin, enabling safer, more consistent manufacturing in cell and gene therapy applications. Optibumin® 25 is the first recombinant 25% human serum albumin for closed-system biomanufacturing, offering consistency, sterility, and GMP compliance. For decades, plasma-derived HSA has presented regulatory and supply chain challenges, including batch variability and pathogen risks. Optibumin 25 eliminates these issues by offering a chemically defined, animal-origin-free, GMP-compliant alternative. It supports regulatory compliance while improving scalability, sterility, and formulation performance. “Optibumin 25 directly replaces plasma-derived 25% HSA while enhancing process control and reducing regulatory risk,” said Scott Deeter, CEO at InVitria. “It enables the kind of consistency required for clinical and commercial-scale production.” Manufactured in an ISO 9001-certified, cGMP-compliant facility, Optibumin 25 is optimized for downstream processing, formulation, and cryopreservation workflows. The product is currently available in 100 mL single-use bags and is suitable for integration into viral vector manufacturing, stem cell preservation, vaccine formulation, and regenerative medicine. Key Benefits: First recombinant 25% HSA designed for closed systems Animal-origin-free and chemically defined GMP-ready and scalable to commercial volumes Stable, sterile, and contamination-resistant To learn more visit: www.invitria.com/optibumin25 About InVitria InVitria develops and manufactures high-performance, animal-origin-free supplements and excipients that support regulatory compliance and product consistency in biologics manufacturing. Its recombinant portfolio enables safer, scalable alternatives to blood-derived materials in cell and gene therapy, vaccine development, and regenerative medicine. InVitria products are manufactured in the United States in cGMP-compliant, ISO 9001-certified facilities.

Ten (10) patients have achieved an objective response (5 confirmed) when treated with MDNA11 alone or in combination with KEYTRUDA® (pembrolizumab) – all responders had advanced and/or metastatic cancers with resistance to immune checkpoint inhibitors (ICI) or with historically low responses to ICI Combination of MDNA11 with KEYTRUDA in the dose escalation arm achieved an objective response rate (ORR) of 36% (5 of 14) in all tumor types enrolling in the Phase 2 expansion cohorts and an ORR of 31% (4 of 13) in cancers planned for the Phase 2 combination expansion cohort Patients from MDNA11 monotherapy dose expansion and dose escalation arms treated at ≥ 60 µg/kg achieved an ORR of 29.4% (5 of 17) in all tumor types enrolling in the Phase 2 expansion cohorts and an ORR of 40% (4 of 10) in cancers enrolling in the Phase 2 monotherapy expansion cohort Highest ORR of 50% achieved amongst MSI-H patients receiving MDNA11 monotherapy (2 of 4) and endometrial cancer patients receiving a combination of MDNA11 and KEYTRUDA (2 of 4) Monotherapy arm continues to demonstrate encouraging durability with a melanoma patient and a pancreatic cancer patient remaining tumor-free and off-treatment after entering the study 2 and 3 years ago, respectively Enrollment in the Phase 2 combination dose expansion arm is underway with the MDNA11 combination Recommended Dose for Expansion (cRDE) established at 90 µg/kg (every 2 weeks) together with KEYTRUDA (400 mg every 6 weeks) Tumor types being evaluated in the Phase 2 expansion cohorts include MSI-H/dMMR, TMB-H, virally-derived cancers (monotherapy), gynecological cancers (combination) and cutaneous melanoma with (combination) or without primary resistance to ICI (monotherapy and combination arms) Additional monotherapy and combination data from the ABILITY-1 study expected at future medical conferences throughout this calendar year TORONTO and HOUSTON, April 28, 2025 (GLOBE NEWSWIRE) -- Medicenna Therapeutics Corp. (“Medicenna” or the “Company”) (TSX: MDNA, OTCQX: MDNAF), a clinical-stage immunotherapy company focused on the development of Superkines targeting cancer and autoimmune diseases, today presents updated clinical data from the ongoing Phase 1/2 ABILITY-1 study evaluating MDNA11 in patients with advanced solid tumors at the 2025 Annual Meeting of the American Association for Cancer Research (AACR) in Chicago, Illinois. MDNA11, a long-acting ‘beta-enhanced not-alpha’ interleukin-2 (IL-2) super-agonist, is being evaluated as a monotherapy or in combination with Merck’s (known as MSD outside of the US and Canada) anti-PD-1 therapy, KEYTRUDA® (pembrolizumab). “We are encouraged by the anti-tumor activity and safety of MDNA11 in both the monotherapy and combination setting, including the 3 recent responders, particularly in patients who have shown resistance to checkpoint inhibitors or with tumor types with historically low responses to immunotherapies and addressing a vastly underserved cancer population,” commented Fahar Merchant, PhD, President and CEO of Medicenna. “Today’s efficacy and immunodynamic data have provided the foundation to commence the dose expansion portion for MDNA11 in combination with KEYTRUDA. In the monotherapy setting, we continue to see deep and durable single agent activity of MDNA11 with two patients remaining in complete remission and off therapy. Together, these data reinforce the best-in-class potential for MDNA11 as a potent and safe immunotherapy that is poised to transform the existing treatment paradigm for patients with difficult-to-treat tumors. We look forward to sharing additional clinical data from the ABILITY-1 study at future medical conferences this year.” Key findings from the on-going ABILITY-1 study (data cut-off as of April 15, 2025) include: Safety Profile MDNA11 continues to demonstrate an acceptable safety profile both as a single agent and in combination with KEYTRUDA. Over 90% of treatment-related adverse events (TRAEs) were Grade 1-2 and transient, with no dose limiting toxicities (DLTs) observed with MDNA11 at doses up to 120 µg/kg (Q2W) in monotherapy and in combination with KEYTRUDA (400 mg, Q6W). Immunodynamics Robust expansion of immune effector cells in monotherapy and combination with KEYTRUDA with notable increases in effector (DNAM), ‘stemness-like’ (TCF-1) and memory CD8+ T cells, which are critical for sustained anti-tumor responses.Robust, dose-dependent lymphocyte expansion in combination with KEYTRUDA, sustained with repeat MDNA11 dosing, and plateauing at the 90 µg/kg MDNA11 dose level. Clinical Activity of MDNA11 in Combination with KEYTRUDA during Dose Escalation Enrollment in the combination dose expansion portion of the ABILITY-1 study is underway having established the MDNA11 combination Recommended Dose for Expansion (cRDE) at 90 µg/kg administered every 2 weeks together with KEYTRUDA administered at 400 mg every 6 weeks. Encouraging anti-tumor activity continues to be observed with MDNA11 in combination with KEYTRUDA in patients enrolled in the dose escalation portion of the study: 5 objective responses (1 CR and 4 PRs) among heavily pre-treated patients, including two previously reported objective responses in historically low immunotherapy responders: 70-year-old male with anal squamous cell carcinoma (SCC) who previously progressed on two prior lines of chemotherapy achieved a CR on the first study evaluable imaging scan (week 8); patient continues on combination treatment following a confirmed CR.52-year-old female with TMB-H/MSS colorectal cancer who previously progressed on two prior lines of chemotherapy achieved a confirmed PR. While off-study for 4 months, the patient continues to show deepening of the tumor response in the absence of any treatment. Objective response rate (ORR) of 31% (4 of 13) in patients with cancers planned for the Phase 2 combination expansion cohort, including cutaneous melanoma, MSI-H/dMMR, and TMB-H tumors, and an ORR of 36% (5 of 14) when including the virally-derived cancers.Stable disease (SD) in 3 patients for a disease control rate (DCR = CR+PR+SD) of 57% (8/14). Monotherapy Tumor Response in Immune Checkpoint Inhibitor-Resistant Patients MDNA11 continues to demonstrate encouraging deep and durable single-agent anti-tumor activity among patients who progressed on prior immune checkpoint inhibitor (ICI) therapy: ORR of 40% in the monotherapy dose escalation (treated at ≥ 60 μg/kg) and expansion arm (1 confirmed CR, 1 confirmed PR and 2 unconfirmed PRs) among 10 patients with tumor types currently enrolling in the Phase 2 monotherapy expansion cohort. All patients had previously failed ICI therapy and had advanced and/or metastatic cutaneous melanoma with secondary resistance to ICI or MSI-H/dMMR tumors. Note that the non-melanoma skin cancer cohort is not recruiting as it was replaced with a new cohort of virally-derived cancers subsequent to a protocol amendment. The ORR is 29.4% from a total of 17 patients when including patients with cancers planned for Phase 2 expansion cohort and patients with primary ICI resistant melanoma who received at least 60 µg/kg of MDNA11 (Q2W).Overall, objective responses in ICI-resistant patients include 1 CR and 4 PRs 2 PRs among 4 MSI-H patients (ORR of 50%) with both responders having metastatic pancreatic ductal adenocarcinoma (PDAC).1 CR and 1 PR among 6 patients with ICI secondary resistant melanoma (ORR of 33.3%).1 PR among patients with ICI primary resistant melanoma. Complete resolution of all target and non-target lesions in two patients with on-going remission following end-of-treatment: Cutaneous melanoma patient achieved a durable response exceeding 12 months during the study with complete resolution of target and non-target metastatic lesions, and continues to remain off anti-cancer therapy for more than 6 weeks.Pancreatic cancer patient (MSI-H) achieved a durable response for 20 months during the study with complete resolution of target and non-target metastatic lesions in the liver including a new lesion treated with MDNA11 following a single cycle of radiation, and continues to remain off anti-cancer therapy for more than 12 months. SD in 6 patients for a disease control rate (DCR = CR+PR+SD) of 65% including 2 with duration > 6 months, yielding a clinical benefit rate of 41% (7/17). A copy of the poster and related slide deck has been posted on the “Scientific Presentations” page of Medicenna’s website. KEYTRUDA® is a registered trademark of Merck Sharp & Dohme LLC, a subsidiary of Merck & Co., Inc., Rahway, NJ, USA. About MDNA11 MDNA11 is an intravenously administered, long-acting, ‘beta-enhanced not-alpha’ IL-2 Superkine specifically engineered to overcome the shortcomings of aldesleukin and other next generation IL-2 variants by preferentially activating immune effector cells (CD8+ T and NK cells) responsible for killing cancer cells, with minimal or no stimulation of immunosuppressive Tregs. These unique proprietary features of the IL-2 Superkine have been achieved by incorporating seven specific mutations and genetically fusing it to a recombinant human albumin scaffold to improve the pharmacokinetic (PK) profile and pharmacological activity of MDNA11 due to albumin’s natural propensity to accumulate in highly vascularized sites, in particular tumor and tumor draining lymph nodes. MDNA11 is currently being evaluated in the Phase 1/2 ABILITY-1 study as both monotherapy and in combination with KEYTRUDA®. About the ABILITY-1 Study The ABILITY-1 study (NCT05086692) is a global, multi-center, open-label study that assesses the safety, tolerability, pharmacokinetics, pharmacodynamics and anti-tumor activity of MDNA11 as monotherapy or in combination with Merck’s anti-PD-1 therapy, KEYTRUDA® (pembrolizumab). In the combination dose escalation portion of the Phase 2 study, patients have been enrolled and administered ascending doses of MDNA11 intravenously in combination with KEYTRUDA. This portion of the study includes patients with a wide range of solid tumors with the potential for susceptibility to immune modulating therapeutics. The combination Recommended Dose for Expansion (cRDE) has been established and the study has commenced combination dose expansion. About Medicenna Therapeutics Medicenna is a clinical-stage immunotherapy company focused on developing novel, highly selective versions of IL-2, IL-4 and IL-13 Superkines and first-in-class Empowered Superkines. Medicenna’s long-acting IL-2 Superkine, MDNA11, is a next-generation IL-2 with superior affinity toward CD122 (IL-2 receptor beta) and no CD25 (IL-2 receptor alpha) binding, thereby preferentially stimulating cancer-killing effector T cells and NK cells. MDNA11 is being evaluated in the Phase 1/2 ABILITY-1 Study (NCT05086692) as a monotherapy and in combination with Merck’s anti-PD-1 therapy, KEYTRUDA® (pembrolizumab). Medicenna’s IL-4 Empowered Superkine, bizaxofusp (formerly MDNA55), has been studied in 5 clinical trials enrolling over 130 patients, including a Phase 2b trial for recurrent GBM, the most common and uniformly fatal form of brain cancer. Bizaxofusp has obtained FastTrack and Orphan Drug status from the FDA and FDA/EMA, respectively. Medicenna’s early-stage high-affinity IL-2β biased IL-2/IL-15 Super-antagonists, from its MDNA209 platform, are being evaluated as potential therapies for autoimmune and graft-versus host diseases. Medicenna’s early-stage BiSKITs™ (Bifunctional SuperKine ImmunoTherapies) and the T-MASK™ (Targeted Metalloprotease Activated SuperKine) programs are designed to enhance the ability of Superkines to treat immunologically “cold” tumors. For more information, please visit www.medicenna.com, and follow us on Twitter and LinkedIn. Forward-Looking Statements This news release may contain forward-looking statements within the meaning of applicable securities laws. Forward-looking statements include, but are not limited to, express or implied statements regarding the future operations of the Company, estimates, plans, strategic ambitions, partnership activities and opportunities, objectives, expectations, opinions, forecasts, projections, guidance, outlook or other statements that are not historical facts, such as statements on the therapeutic treatment potential and safety profile of MDNA11 (both as monotherapy and in combination with Merck’s anti-PD-1 therapy, KEYTRUDA® [pembrolizumab]) and the timing and/or release of any additional clinical updates. Drug development and commercialization involve a high degree of risk, and only a small number of research and development programs result in commercialization of a product. Results in early-stage pre-clinical or clinical studies may not be indicative of full results or results from later stage or larger scale clinical studies and do not ensure regulatory approval. You should not place undue reliance on these statements, or the scientific data presented. Forward-looking statements are often identified by terms such as “will”, “may”, “should”, “anticipate”, “expect”, “believe”, “seek”, “potentially” and similar expressions. and are subject to risks and uncertainties. Forward-looking statements are based on a number of assumptions believed by the Company to be reasonable at the date of this news release. Although the Company believes that the expectations reflected in such forward-looking statements are reasonable, there can be no assurance that such statements will prove to be accurate. These statements are subject to certain risks and uncertainties and may be based on assumptions that could cause actual results and future events to differ materially from those anticipated or implied in such statements. Important factors that could cause actual results to differ materially from the Company’s expectations include the risks detailed in the latest annual information form of the Company and in other filings made by the Company with the applicable securities regulators from time to time in Canada. The reader is cautioned that assumptions used in the preparation of any forward-looking information may prove to be incorrect. Events or circumstances may cause actual results to differ materially from those predicted, as a result of numerous known and unknown risks, uncertainties, and other factors, many of which are beyond the control of the Company. The reader is cautioned not to place undue reliance on any forward-looking information. Such information, although considered reasonable by management, may prove to be incorrect and actual results may differ materially from those anticipated or implied in forward-looking statements. Forward-looking statements contained in this news release are expressly qualified by this cautionary statement. The forward-looking statements contained in this news release are made as of the date hereof and except as required by law, we do not intend and do not assume any obligation to update or revise publicly any of the included forward-looking statements. This news release contains hyperlinks to information that is not deemed to be incorporated by reference in this new release. Investor and Company Contact: Christina CameronInvestor Relationsir@medicenna.com(647) 953-0673

OTC2025论坛深度聚焦类器官与疾病建模、新药发现/研发、3D细胞培养、类器官培养及质控。展位咨询请联系：王晨 180 1628 8769。在《Nature》期刊上发表的这篇文章中，日本的科研团队探讨了通过激活Wnt和STAT3信号通路来实现人类成年肝细胞类器官的长期自我更新。这项研究解决了肝细胞在体外培养中扩增能力与功能保持之间的权衡问题。研究发现，YAP的激活促进了肝细胞的增殖，但会导致其向胆管导管谱系的分化。相反，OSM诱导的STAT3激活不仅促进了肝细胞的增殖，还抑制了导管化生，维持了肝细胞的特性。移植到小鼠体内的肝细胞类器官能够重新构建受体小鼠肝脏的代谢分区结构，并在去除培养基中的特定因子后形成具有胆管网络的绳索状结构，表现出与体内肝细胞相当的主要肝脏代谢功能。该研究为开发针对人类肝脏疾病的治疗策略提供了新的途径。01研究背景首先，肝脏是人体内一个关键的代谢器官，负责多种重要的生物化学过程，如葡萄糖生成、糖原合成、尿素循环、脂质生成、胆汁酸合成、蛋白质生产和药物代谢。这些功能的严重损害会导致肝功能衰竭，通常需要进行肝移植。近年来，随着抗病毒治疗的进步，肝功能衰竭的病因从病毒性肝炎转向与现代生活方式相关的代谢性肝病，特别是代谢功能障碍相关的脂肪性肝病。这种疾病如果不加以治疗，会进展为脂肪性肝炎、肝硬化和肝功能衰竭。然而，由于缺乏能够真实再现肝脏代谢功能的人类肝细胞模型，预防代谢性肝病的成功率有限。其次，体外分离的肝细胞在增殖能力上迅速下降，与其在体内的强大再生能力形成对比。现有的培养方法只能暂时维持初级人类肝细胞的功能性，但会影响其扩增能力。虽然一些研究尝试通过不同的策略来克服这一限制，如使用小分子抑制剂促进肝细胞的扩增，但这些方法往往导致胆管化生，即功能性细胞被重新编程为增殖能力较强的细胞。这种现象被称为“paligenosis”，使得在再生医学、药物发现和毒理学领域中使用人类肝细胞面临重大挑战。因此，开发一种能够长期扩增且保持功能性的人类肝细胞培养系统至关重要。02研究发现人类成年肝细胞类器官的长期扩增和功能保持：研究团队通过激活Wnt和STAT3信号通路，实现了人类成年肝细胞类器官的长期自我更新。YAP信号的激活促进了肝细胞的增殖，但会导致其向胆管细胞谱系分化。相反，Oncostatin M通过激活STAT3信号，促进了肝细胞的增殖，同时抑制了胆管化生，保持了肝细胞的特性。研究表明，这种类器官在移植到小鼠体内后，能够重建受体小鼠肝脏的代谢分区结构，并在去除特定培养因子后，形成具有胆小管网络的肝索样结构，表现出与体内肝细胞相当的主要肝脏代谢功能。类器官在基因编辑和疾病建模中的应用潜力：研究表明，这些肝细胞类器官可以进行基因编辑，从而为代谢性肝病的功能建模提供了可能性。新开发的培养系统为人类肝病的治疗策略开发提供了一个有前景的途径。通过优化的培养条件，研究团队成功地在体外模拟了肝细胞的代谢功能，这标志着功能性类器官医学的新时代。03临床意义肝移植的替代方案：HHOs可能成为肝移植的替代方案，特别是在供体短缺或无法进行肝移植的情况下。  药物筛选和毒性测试：HHOs展示的药物代谢能力与体内肝细胞相似，使其成为药物筛选和毒性测试的理想模型。这可以加速新药的开发和安全性评估。  代谢性肝病的研究和治疗开发：HHOs提供了一个真实反映人类肝脏代谢功能的平台，有助于深入研究代谢性肝病的病理机制，并开发针对这些疾病的治疗策略。  再生医学的应用：由于HHOs能够进行基因编辑，它们可用于研究遗传性肝病并探索基因治疗的可行性。  总之，该研究不仅为肝细胞的体外长时间培养提供了新的策略，还为肝脏相关疾病的基础研究和临床应用开辟了新的方向。04实验策略1. 信号通路激活与抑制：  通过激活Wnt和STAT3信号通路，实现人类成年肝细胞类器官的长期自我更新。 YAP激活促进肝细胞增殖，但同时导致其向胆管导向分化。 通过抑制TGFβ信号和使用Oncostatin M激活STAT3，促进肝细胞增殖并抑制导管化生，从而保持肝细胞特性。2. 类器官培养体系的优化：  在培养基中添加EGF、HGF、FGF-10、Wnt/R-spondin、TGFβ抑制剂（A83-01和Noggin）、IL-6和PKA激活剂（Forskolin），支持小囊状类器官的形成。 通过去除某些培养因子（如Noggin和forskolin），并补充激素（如生长激素、催乳素和皮质醇），促进肝细胞类器官的线索状结构形成及胆小管网络的建立。3. 体内移植实验：  将类器官移植到经过药物处理以去除肝细胞的小鼠体内，评估其在小鼠肝脏中的再生能力和代谢分区结构的重建。4. 基因编辑与功能建模：  类器官易于进行基因编辑，从而用于代谢性肝病的功能建模。05数据解读图1：人类成人肝细胞类器官在特定因子下的扩增Figure 1 展示了人类成人肝细胞类器官（HHOs）在特定培养条件下的扩增情况，包括使用不同因子如IL-6和OSM对类器官的影响。  A. 为了观察PHH来源的类器官在扩增培养基（加IL-6）中的表达情况，作者进行了免疫荧光染色。结果显示，HHOs表达白蛋白，而ICOs则显示CK19和EpCAM的表达。此结果在三个供体中获得一致性。  B. 为了研究HHOs在扩增培养基中添加OSM或IL-6后的扩增情况，作者通过传代比率和基于ATP的细胞计数估算了细胞数量。每个点代表一次传代事件，三个类器官系在初始3个月内大约每2周传代一次，传代比率为3-12倍，‘x’表示传代失败。供体分别为39岁、53岁和61岁。  C. 为了观察OSM对单个PHH来源类器官生长的影响，作者拍摄了时间推移图像。相同的类器官在相同放大倍率下显示。此结果在两个供体中获得一致性。  D. 为了研究OSM对HHOs基底极性的影响，作者进行了代表性明场和整体免疫染色，标记了CDH1（蓝色）、ZO-1（红色）、整合素β4（绿色）和细胞核（白色）。此结果在四个供体中获得一致性。  E. 为了分析胆管细胞相关转录因子、YAP靶基因和TGFβ信号相关基因的转录组，作者对PHHs、ICOs和HHOs在有无OSM和LATSi（TRULI，5 μM）条件下进行了分析。供体分别为11个月和16个月。  F. 为了观察HHOs在±OSM和±LATSi（5 μM）培养14天后的情况，作者进行了Alb和CK19的免疫染色（左）。细胞核用Hoechst 33342标记。结果显示，CK19+类器官仅在无OSM和有LATSi的条件下观察到（右）。  结论：人类成人肝细胞类器官在特定因子如IL-6和OSM的影响下能够有效扩增，并且不同因子组合对类器官的极性和表型有显著影响。图2：小鼠肝脏用人类肝类器官（HHO）再生Figure 2 展示了通过异种移植方法，将人类肝类器官（HHO）移植到小鼠肝脏中，观察其在小鼠肝脏中的再生情况。  a. 为了研究HHO在小鼠肝脏中的再生能力，作者设计了异种移植实验流程。TK-NOG小鼠在接受ValGCV处理以消融肝细胞后，被移植了HHO或人类原代肝细胞（PHH）。每周采集血液样本，并在8周后收集肝脏样本。  b. 通过苏木精-伊红染色（H&E）和STEM121染色，展示了HHO移植物的代表性图像。结果显示，移植的小鼠和人类肝细胞之间有明显的界限。类似结果在21个PHH移植物（来自五个供体）和104个HHO移植物（来自八个供体）中获得。  c–h. 研究了PHH或HHO（来自两个供体）在小鼠肝脏中的再生情况。通过测量血清中人类白蛋白（hALB）的浓度随时间的变化来评估再生效果。不同的HHO传代次数用不同颜色表示；‘x’表示小鼠死亡。STEM121染色显示了PY53和PY30 HHO移植物。再生效率通过人类肝细胞面积相对于总肝脏面积的比例计算。每个点代表一只小鼠。统计分析显示，与对照组PHH相比，HHO组的再生效率显著提高。  i. 对HHO移植物进行了hALB（红色）、闭合蛋白（绿色）和人类MRP2（蓝色）的染色。结果显示，来自PHH（一个供体）和eHHO（三个供体）的数据具有相似的结果。  j. 在HHO移植物中对分区标记物进行染色，HAL（红色；顶部）位于门静脉附近，CYP2E1（红色；底部）位于中央静脉附近。细胞核用Hoechst 33342（白色）染色；移植细胞为STEM121+（绿色）。门静脉和中央静脉的腔用虚线标出（*表示门静脉）。来自PHH（两个供体）和eHHO（HAL为六个供体，CYP2E1为八个供体）的代表性数据显示了相似的结果。  k,l. 分别显示了在STEM121+区域内HAL+（k）或CYP2E1+（l）面积的比例。每个点代表一只小鼠的一个切片；每种条件下来自两到三只小鼠的平均比例。使用Welch双侧t检验进行统计分析。  结论：HHO能够在小鼠肝脏中有效再生，并表现出与人类肝细胞相似的功能特性。图3：建立具有代谢功能的dHHOs（分化后的人体肝细胞类器官）Figure 3 展示了文章中的一项关键实验，即分化后的人体肝细胞类器官（dHHO）建立及其代谢功能的评估。  形态变化： dHHOs从上皮样结构转变为肝细胞样结构，显示出成熟的形态特征。 胆汁酸代谢： 实验证明dHHOs可以合成胆汁酸，并在胆汁小管中运输荧光标记的胆汁酸衍生物，表明其具有生理功能。 胆汁酸的合成和分泌： LC–MS/MS分析显示，与体外培养的人体肝细胞相比，dHHOs产生更高浓度的胆汁酸。 甘油三酯生产： dHHOs能够合成甘油三酯，并在存在微粒体甘油三酯传输蛋白抑制剂时被阻断，验证了其代谢活性。 结论： 这些成功的分化实验及其结果表明，通过优化培养条件，dHHOs不仅展示了典型的胆汁酸代谢功能，还具备了潜在的药物发现和毒理学研究的应用能力。这为将来利用类器官模型进行更深层次的人类肝脏代谢研究奠定了基础。图4：建立门静脉肝细胞类器官Figure 4 展示了对分化后的人体肝细胞类器官（dHHOs）的功能评估，特别是在再现肝脏门静脉区域特定功能方面的能力。  葡萄糖合成与糖异生： dHHOs在饥饿后展示了较高的葡萄糖生成能力，并成功追踪到13C标记的葡萄糖合成路径，证实了其有效的糖异生能力。 尿素合成： 使用15N标记氯化铵进行追踪，发现dHHOs能够转变氨为尿素，验证了其尿素循环的完整性。 线粒体功能活性： dHHOs显示出高于eHHOs的最大呼吸能力，提示其具有更活跃的氧化磷酸化水平。 白蛋白和凝血因子生产： dHHOs能够合成接近人体生理水平的白蛋白，并在维生素K存在下增加凝血因子的活性。 结论： 通过一系列功能性实验，Figure 4展示了dHHOs在多种重要肝脏代谢功能方面的恢复能力，特别是与门静脉区域相关的功能。这包括葡萄糖生成、尿素合成及相应代谢途径的活跃性，表明dHHOs可以作为体外研究人类肝脏功能和代谢疾病的有效模型。图5：生成基因敲除的肝细胞类器官Figure 5 展示了通过基因编辑技术生成基因敲除的人体肝细胞类器官（HHOs）的过程和结果。  基因敲除成功： 成功通过sgRNA实现了对G6PC和OTC基因的敲除，Sanger测序和免疫检测确认了对应蛋白的缺失。 形态和功能保持： 尽管基因敲除，G6PC和OTC缺失的HHOs仍然保持了正常的类器官形态，并能够合成一定水平的白蛋白。 功能影响： G6PC敲除使得糖异生功能降低，而OTC敲除导致瓜氨酸生成的缺失，验证了各基因在其代谢途径中的关键角色。 培养条件优化： 通过使用LATSi和人血小板裂解物，促进了基因编辑HHOs的扩增，而没有引起生长停滞或管道分化。 结论： Figure 5描述了一种有效的流程来对成人肝细胞类器官进行基因敲除，从而提供一种稳定的模型来研究肝脏代谢疾病。这一进展为通过基因编辑技术探索肝脏功能、发现疾病机理和开发治疗策略提供了可能性。06主要结论这项研究展示了一种新的培养系统，该系统能够在体外长期扩增人类成体肝细胞类器官（HHOs），并保持其代谢功能。通过激活Wnt和STAT3信号通路，研究人员成功实现了HHOs的自我更新，并避免了导管化生的发生。HHOs能够重新构建受体小鼠肝脏的代谢分区结构，并在体外展示出与体内肝细胞相当的主要肝脏代谢功能。这种文化系统为肝脏疾病的治疗策略开发提供了新的途径。07讨论总结研究强调了HHOs技术在解决人类成体肝细胞体外扩增与功能性之间权衡的潜力。通过基因组学分析，关键的培养因子如EHF、Wnt/R-spondin、Noggin、TGFβ抑制剂、OSM和福司可林被确定为促进肝细胞扩增的必要因素。特别地，OSM-STAT3激活不仅促进了肝细胞的增殖，还保留了肝脏谱系的完整性，防止了导管化生和上皮-间质转化。  研究指出，HHOs技术代表了一种新的类器官培养系统，兼具肝细胞的扩增性和功能性。其优点包括提供了一个稳定、可重复且成本效益高的平台，用于探索人类肝脏代谢，超越了传统的人类肝细胞模型的局限性。此外，HHOs在再生医学中的潜在应用为解决肝衰竭问题打开了新的视野。END免责声明：本文仅作知识交流与分享及科普目的，不涉及商业宣传，不作为相关医疗指导或用药建议。文章如有侵权请联系删除。OTC2025论坛深度聚焦类器官与疾病建模、新药发现/研发、3D细胞培养、类器官培养及质控。展位咨询请联系：王晨 180 1628 8769。戳“阅读原文”立即领取OTC2025免费参会名额!