Immunosuppressed Miniswine as a Model for Testing Cell Therapy Success: Experience With Implants of Human Salivary Stem/Progenitor Cell Constructs.
Immunosuppressed Miniswine as a Model for Testing Cell Therapy Success: Experience With Implants of Human Salivary Stem/Progenitor Cell Constructs.
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DOI：
10.3389/fmolb.2021.711602
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发表时间：
2021
影响因子：
5
通讯作者：
Farach-Carson MC
中科院分区：
文献类型：
--
作者：
Wu D;Lombaert IMA;DeLeon M;Pradhan-Bhatt S;Witt RL;Harrington DA;Trombetta MG;Passineau MJ;Farach-Carson MC
迫切需要开发大型动物模型，用于临床前测试新的细胞疗法，以取代丢失或受损的组织。接受放射治疗头颈部癌症的患者经常出现口干/口干，这种情况有朝一日可能会通过细胞疗法来重新填充功能强大的唾液产生细胞。使用为接受全面部移植的患者开发的免疫抑制方案，我们成功地使用免疫抑制的迷你葡萄酒作为合适的宿主动物来评价基质修饰的透明质酸(HA)水凝胶/生物支架材料的长期稳定性、生物相容性和命运，该材料含有包裹的唾液干细胞/祖细胞(HS/PC)。在未经处理的迷你葡萄酒的腮腺中进行了初步的生物相容性测试。随后在动物身上进行了使用HS/PC水凝胶的实验，在手术当天开始了免疫抑制方案。植入部位包括用于活性测试的肾胶囊和用于生物整合时间长达8周的腮腺。通过分析植入部位附近的组织，在任何动物身上都没有观察到移植排斥反应。事实证明，第一代植入物只包含水凝胶中的细胞，在手术室中很难处理，经过修改后，可以附着在猪小肠粘膜下层(SIS)膜上，以改进处理能力，并可以通过达芬奇手术系统进行输送。使用第二代3D-唾液组织(3D-ST)评估了几种不同的手术技术在肾囊和无囊腮腺上的易用性和稳定性。对于肾脏，将植入物滑动到囊膜下，快速缝合优于其他方法。对于腮腺，在组织损伤最小的情况下，可以很好地耐受用于放置的组织“口袋”和立即闭合多层组织。手术夹被放置作为组织采集的受托标记。在照射后90天，当唾液明显减少时，用迷你葡萄酒进行了一些植入实验。仍有足够的腮腺组织可供植入，动物可耐受免疫抑制。在所有实验中，植入的HS/PC的存活率很高，腮腺植入物中有明显的血管和神经系统整合的迹象。因此，我们得出结论，免疫抑制的迷你葡萄酒是在第一次人体试验之前测试人类植入物的一种高价值的新兴模型。
An urgent need exists to develop large animal models for preclinical testing of new cell therapies designed to replace lost or damaged tissues. Patients receiving irradiation for treatment of head and neck cancers frequently develop xerostomia/dry mouth, a condition that could one day be treated by cell therapy to repopulate functional saliva-producing cells. Using immunosuppression protocols developed for patients receiving whole face transplants, we successfully used immunosuppressed miniswine as a suitable host animal to evaluate the long-term stability, biocompatibility, and fate of matrix-modified hyaluronate (HA) hydrogel/bioscaffold materials containing encapsulated salivary human stem/progenitor cells (hS/PCs). An initial biocompatibility test was conducted in parotids of untreated miniswine. Subsequent experiments using hS/PC-laden hydrogels were performed in animals, beginning an immunosuppression regimen on the day of surgery. Implant sites included the kidney capsule for viability testing and the parotid gland for biointegration time periods up to eight weeks. No transplant rejection was seen in any animal assessed by analysis of the tissues near the site of the implants. First-generation implants containing only cells in hydrogel proved difficult to handle in the surgical suite and were modified to adhere to a porcine small intestinal submucosa (SIS) membrane for improved handling and could be delivered through the da Vinci surgical system. Several different surgical techniques were assessed using the second-generation 3D-salivary tissue (3D-ST) for ease and stability both on the kidney capsule and in the capsule-less parotid gland. For the kidney, sliding the implant under the capsule membrane and quick stitching proved superior to other methods. For the parotid gland, creation of a tissue “pocket” for placement and immediate multilayer tissue closure were well tolerated with minimal tissue damage. Surgical clips were placed as fiduciary markers for tissue harvest. Some implant experiments were conducted with miniswine 90 days post-irradiation when salivation decreased significantly. Sufficient parotid tissue remained to allow implant placement, and animals tolerated immunosuppression. In all experiments, viability of implanted hS/PCs was high with clear signs of both vascular and nervous system integration in the parotid implants. We thus conclude that the immunosuppressed miniswine is a high-value emerging model for testing human implants prior to first-in-human trials.