Department of Mechatronics Engineering, College of Engineering, Incheon National University, 119 Academy-ro, Yeonsu-gu, Incheon, 22012, Republic of Korea.
3D Stem Cell Bioengineering Laboratory, Research Institute for Engineering and Technology, Incheon National University, Incheon, 22012, Republic of Korea.
Tissue Eng Regen Med. 2022 Aug;19(4):739-754. doi: 10.1007/s13770-022-00447-3. Epub 2022 May 9.
As stem cells are considered a promising cell source for tissue engineering, many culture strategies have been extensively studied to generate in vitro stem cell-based tissue constructs. However, most approaches using conventional tissue culture plates are limited by the lack of biological relevance in stem cell microenvironments required for neotissue formation. In this study, a novel perfusion rotating wall vessel (RWV) bioreactor was developed for mass-production of stem cell-based 3D tissue constructs.
An automated RWV bioreactor was fabricated, which is capable of controlling continuous medium perfusion, highly efficient gas exchange with surrounding air, as well as low-intensity pulsed ultrasound (LIPUS) stimulation. Embryonic stem cells encapsulated in alginate/gelatin hydrogel were cultured in the osteogenic medium by using our bioreactor system. Cellular viability, growth kinetics, and osteogenesis/mineralization were thoroughly evaluated, and culture media were profiled at real time. The in vivo efficacy was examined by a rabbit cranial defect model.
Our bioreactor successfully maintained the optimal culture environments for stem cell proliferation, osteogenic differentiation, and mineralized tissue formation during the culture period. The mineralized tissue constructs produced by our bioreactor demonstrated higher void filling efficacy in the large bone defects compared to the group implanted with hydrogel beads only. In addition, the LIPUS modules mounted on our bioreactor successfully reached higher mineralization of the tissue constructs compared to the groups without LIPUS stimulation.
This study suggests an effective biomanufacturing strategy for mass-production of implantable mineralized tissue constructs from stem cells that could be applicable to future clinical practice.
由于干细胞被认为是组织工程有前途的细胞来源，因此已经广泛研究了许多培养策略，以在体外生成基于干细胞的组织构建体。然而，大多数使用传统组织培养板的方法都受到限制，因为缺乏新组织形成所需的干细胞微环境的生物学相关性。在这项研究中，开发了一种新型灌注旋转壁式（RWV）生物反应器，用于大规模生产基于干细胞的 3D 组织构建体。
制造了一种自动化的 RWV 生物反应器，该生物反应器能够控制连续的培养基灌注，与周围空气进行高效的气体交换，以及低强度脉冲超声（LIPUS）刺激。将藻酸盐/明胶水凝胶包封的胚胎干细胞在成骨培养基中使用我们的生物反应器系统进行培养。彻底评估了细胞活力、生长动力学和成骨/矿化，并且实时分析了培养介质。通过兔颅缺损模型检查了体内功效。
我们的生物反应器在培养期间成功维持了干细胞增殖、成骨分化和矿化组织形成的最佳培养环境。与仅植入水凝胶珠的组相比，我们的生物反应器生产的矿化组织构建体在大骨缺损中具有更高的空隙填充效果。此外，安装在我们的生物反应器上的 LIPUS 模块成功达到了比没有 LIPUS 刺激的组更高的组织构建体矿化程度。
这项研究提出了一种有效的生物制造策略，用于大规模生产可植入的矿化组织构建体，该策略可适用于未来的临床实践。
