Department of Electrical and Computer Engineering, College of Engineering, Texas A&M University, College Station, TX 77843-3128, USA.
Biomed Microdevices. 2009 Dec;11(6):1145-53. doi: 10.1007/s10544-009-9331-7.
This paper presents a circular microfluidic compartmentalized co-culture platform that can be used for central nervous system (CNS) axon myelination research. The microfluidic platform is composed of a soma compartment and an axon/glia compartment connected through arrays of axon-guiding microchannels. Myelin-producing glia, oligodendrocytes (OLs), placed in the axon/glia compartment, interact with only axons but not with neuronal somata confined to the soma compartment, reminiscent to in vivo situation where many axon fibres are myelinated by OLs at distance away from neuronal cell bodies. Primary forebrain neurons from embryonic day 16-18 rats were cultured inside the soma compartment for two weeks to allow them to mature and form extensive axon networks. OL progenitors, isolated from postnatal day 1-2 rat brains, were then added to the axon/glia compartment and co-cultured with neurons for an additional two weeks. The microdevice showed fluidic isolation between the two compartments and successfully isolated neuronal cell bodies and dendrites from axons growing through the arrays of axon-guiding microchannels into the axon/glia compartment. The circular co-culture device developed here showed excellent cell loading characteristics where significant numbers of cells were positioned near the axon-guiding microchannels. This significantly increased the probability of axons crossing these microchannels as demonstrated by the more than 51 % of the area of the axon/glia compartment covered with axons two weeks after cell seeding. OL progenitors co-cultured with axons inside the axon/glia compartment successfully differentiated into mature OLs. These results indicate that this device can be used as an excellent in vitro co-culture platform for studying localized axon-glia interaction and signalling.
本文提出了一种环形微流控分隔共培养平台，可用于中枢神经系统（CNS）轴突髓鞘形成研究。该微流控平台由胞体腔室和轴突/胶质腔室组成，通过轴突导向微通道阵列连接。将产生髓鞘的胶质细胞（少突胶质细胞）置于轴突/胶质腔室内，它们仅与轴突相互作用，而不与局限于胞体腔室内的神经元胞体相互作用，这种情况类似于体内许多轴突纤维由远离神经元胞体的胶质细胞在远处进行髓鞘化的情况。取胚胎 16-18 天大鼠的原代前脑神经元培养在胞体腔室内两周，使其成熟并形成广泛的轴突网络。然后从出生后 1-2 天大鼠的大脑中分离出少突胶质细胞前体细胞，并添加到轴突/胶质腔室内，与神经元共培养两周。该微器件在两个腔室之间显示出流体隔离，并成功地将神经元胞体和树突与穿过轴突导向微通道阵列进入轴突/胶质腔室的轴突分离。所开发的环形共培养装置具有出色的细胞加载特性，大量细胞被定位在轴突导向微通道附近。这显著增加了轴突穿过这些微通道的可能性，如在细胞接种两周后，超过 51%的轴突/胶质腔室面积被轴突覆盖。与轴突共培养的少突胶质细胞前体细胞在轴突/胶质腔室内成功分化为成熟的少突胶质细胞。这些结果表明，该装置可用作研究局部轴突-胶质相互作用和信号传递的优秀体外共培养平台。
