Warburton Linnea, Rubinsky Boris
Department of Mechanical Engineering, University of California Berkeley, Berkeley, CA 94720, USA.
Department of Bioengineering, University of California Berkeley, Berkeley, CA 94720, USA.
Gels. 2023 Jun 20;9(6):502. doi: 10.3390/gels9060502.
Temperature-Controlled-Cryoprinting (TCC) is a new 3D bioprinting technology that allows for the fabrication and cryopreservation of complex and large cell-laden scaffolds. During TCC, bioink is deposited on a freezing plate that descends further into a cooling bath, keeping the temperature at the nozzle constant. To demonstrate the effectiveness of TCC, we used it to fabricate and cryopreserve cell-laden 3D alginate-based scaffolds with high cell viability and no size limitations. Our results show that Vero cells in a 3D TCC bioprinted scaffold can survive cryopreservation with a viability of 71%, and cell viability does not decrease as higher layers are printed. In contrast, previous methods had either low cell viability or decreasing efficacy for tall or thick scaffolds. We used an optimal temperature profile for freezing during 3D printing using the two-step interrupted cryopreservation method and evaluated drops in cell viability during the various stages of TCC. Our findings suggest that TCC has significant potential for advancing 3D cell culture and tissue engineering.
温度控制冷冻打印（TCC）是一种新型的3D生物打印技术，可用于制造和冷冻保存复杂的大型载细胞支架。在TCC过程中，生物墨水沉积在一块进一步下降到冷却浴中的冷冻板上，从而保持喷嘴处的温度恒定。为了证明TCC的有效性，我们用它制造并冷冻保存了具有高细胞活力且无尺寸限制的基于海藻酸盐的载细胞3D支架。我们的结果表明，3D TCC生物打印支架中的Vero细胞在冷冻保存后存活率可达71%，且随着打印层数增加细胞活力并未下降。相比之下，以前的方法要么细胞活力低，要么对于高大或厚实的支架效果会降低。我们在3D打印过程中使用两步间断冷冻保存法采用了最佳的冷冻温度曲线，并评估了TCC各个阶段的细胞活力下降情况。我们的研究结果表明，TCC在推进3D细胞培养和组织工程方面具有巨大潜力。
