Tendulkar S, McQuilling J P, Childers C, Pareta R, Opara E C, Ramasubramanian M K
Department of Mechanical and Aerospace Engineering, North Carolina State University, Raleigh, North Carolina 27695-7910, USA.
Transplant Proc. 2011 Nov;43(9):3184-7. doi: 10.1016/j.transproceed.2011.10.023.
The objective of this research was to test the viability and function of islets microencapsulated using a scalable microfluidic device that is suitable for the mass production of encapsulated islets for transplantation. A 3-D microfluidic device consisting of eight outlets with an inner fluid inlet and an outer concentric inlet to the device has been designed and fabricated using the stereolithography rapid prototyping technique. Islets were isolated from normal Wistar-Furth rat pancreas using the procedure of collagenase digestion of pancreatic tissue. Following purification, islet suspensions in 1.5% sodium alginate were pumped into the fluid inlet of the microfluidic device, which distributed the flow equally to all the eight channels according to the design. The air plenum distributed compressed air uniformly through the eight concurrent outlets, and with one fluid pump and air source, the device produced eight microencapsulations simultaneously. After encapsulation, the islets were tested for functionality using the dynamic perifusion procedure with low- and high-glucose concentrations. The device is capable of producing eight channels of steady stream of monodisperse microencapsulations of a range of diameters depending on the design and process parameters. Using this prototype device, encapsulated islets were shown to be viable in the functional tests that we performed. Thus, the mean ± standard deviation rate of insulin secretion increased from a basal rate of 0.165 ± 0.059 ng/10 islets/min to a stimulated rate of 0.422 ± 0.095 ng/10 islets/min (P < .05, n = 3), when the glucose concentration was changed from 5.5 mmol/L to 27.5 mmol/L, and this glucose stimulation index was not different from that observed with unencapsulated islets under same conditions. In summary, the high-throughput prototype device that we have designed can produce encapsulated islets that are viable and suitable for transplantation studies.
本研究的目的是测试使用可扩展微流控装置微囊化的胰岛的活力和功能,该装置适用于大规模生产用于移植的微囊化胰岛。利用立体光刻快速成型技术设计并制造了一种三维微流控装置,该装置由八个出口、一个内部流体入口和一个与装置同心的外部入口组成。采用胶原酶消化胰腺组织的方法从正常Wistar-Furth大鼠胰腺中分离胰岛。纯化后,将1.5%海藻酸钠中的胰岛悬浮液泵入微流控装置的流体入口,该入口根据设计将流体均匀分配到所有八个通道。空气腔通过八个并行出口均匀分布压缩空气,通过一个流体泵和一个空气源,该装置可同时进行八个微囊化操作。微囊化后,使用低葡萄糖浓度和高葡萄糖浓度的动态灌流程序对胰岛的功能进行测试。根据设计和工艺参数,该装置能够产生八个通道的稳定单分散微囊流,其直径范围不同。使用该原型装置,在我们进行的功能测试中,微囊化胰岛显示出具有活力。因此,当葡萄糖浓度从5.5 mmol/L变为27.5 mmol/L时,胰岛素分泌的平均±标准差率从基础率0.165±0.059 ng/10个胰岛/分钟增加到刺激率0.422±0.095 ng/10个胰岛/分钟(P<0.05,n = 3),并且该葡萄糖刺激指数与在相同条件下未微囊化胰岛所观察到的指数没有差异。总之,我们设计的高通量原型装置能够生产出具有活力且适合移植研究的微囊化胰岛。