Long-term immunologically competent human peripheral lymphoid tissue cultures in a 3D bioreactor.
Long-term immunologically competent human peripheral lymphoid tissue cultures in a 3D bioreactor.
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在 3D 生物反应器中长期培养具有免疫活性的人外周淋巴组织。
DOI：
10.1002/bit.23055
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发表时间：
2011
影响因子：
3.8
通讯作者：
Bottaro,Andrea
中科院分区：
文献类型：
--
作者：
Kuzin,Igor;Sun,Hongliang;Moshkani,Safiekhatoon;Feng,Changyong;Mantalaris,Athanasios;Wu,JHDavid;Bottaro,Andrea
外周淋巴器官（PLO），适应性免疫应答发展的主要部位，显示出反映细胞亚群分离的复杂结构组织（例如，T和B淋巴细胞）和对免疫功能至关重要的功能区室。在体外培养系统的产生，能够概括PLO的实验，生物技术和临床应用的显着特点将是非常可取的，但迄今为止，这些系统的复杂性受到阻碍。我们之前已经开发了一种三维生物反应器系统，用于在填充床生物反应器中的大孔微球上长期功能性培养人骨髓细胞，并经常更换培养基。在这里，我们适应相同的系统培养人原代细胞从PLO（扁桃体）在特定的外源性生长因子或激活剂的情况下。该系统中的细胞在几周内显示出更高的活力，并保持了与原代细胞基本相当的群体多样性和细胞表面标志物。光学显微镜检查显示，细胞在支架孔内组织成大的不同簇，并存在B细胞富集区。引人注目的是，当用一组不同抗原进行攻击时，这些培养物产生了大量产生抗体的B细胞，正如淋巴组织所预期的那样。因此，三维扁桃体生物反应器培养系统可以通过在体外重现其结构组织和功能而用作PLO的有用模型。Biotechnol. Bioeng. 2011; 108：1430 - 1440.© 2011 Wiley Periodicals，Inc.
Peripheral lymphoid organs (PLOs), the primary sites of development of adaptive immune responses, display a complex structural organization reflecting separation of cellular subsets (e.g., T and B lymphocytes) and functional compartments which is critical for immune function. The generation of in vitro culture systems capable of recapitulating salient features of PLOs for experimental, biotechnological, and clinical applications would be highly desirable, but has been hampered so far by the complexity of these systems. We have previously developed a three‐dimensional bioreactor system for long‐term, functional culture of human bone marrow cells on macroporous microspheres in a packed‐bed bioreactor with frequent medium change. Here we adapt the same system for culture of human primary cells from PLOs (tonsil) in the absence of specific exogenous growth factors or activators. Cells in this system displayed higher viability over several weeks, and maintain population diversity and cell surface markers largely comparable to primary cells. Light microscopy showed cells organizing in large diverse clusters within the scaffold pores and presence of B cell‐enriched areas. Strikingly, these cultures generated a significant number of antibody‐producing B cells when challenged with a panel of diverse antigens, as expected from a lymphoid tissue. Thus the three‐dimensional tonsil bioreactor culture system may serve as a useful model of PLOs by recapitulating their structural organization and function ex vivo. Biotechnol. Bioeng. 2011; 108:1430–1440. © 2011 Wiley Periodicals, Inc.