Cultured neurons, like the neurons that constitute the in vivo brain, are capable of expressing ion channels capable of generating action potentials, observing the growth of axons and dendrites, and forming neurons and synapses. A neural network consisting of cultured neural networks is used as a bottom-up approach to study the brain. The cultured neural network is reduced as compared to the neural network of the in vivo brain, and it is easy to use as a model because it has the basic electrophysiological characteristics of the brain neural network. Since it is easy to regulate the growth of cultured neurons externally, studies are underway to control the topology of the networks formed from cultured neurons. Surface patterning techniques can be used to control the neural adhesion and neurite outgrowth of in vitro culture substrates at cellular scale. This approach basically defines cell adhesive region and cell repellent region on the culture substrate by using surface patterning and realizes cell adhesion and neurite outgrowth only in relatively cell adhesive region to make neural network of desired design. This dissertation deals with the Soft-lithography based surface micro-patterning approaches for controlling the topology of in vitro neural networks. One of the soft lithography technologies, microcontact printing technology, was used to pattern the biomolecule with a microscale, and micromolding was used to pattern the cell repellent materials agarose hydrogel and alginate hydrogel to design the neural network. The physiological characteristics of the neural network formed by the micropatterning technique were examined by using immunohistochemistry, and the electrical signals were measured from the designed neural network using the microelectrode chip system, and the electrophysiological characteristics and connectivity were analyzed. This dissertation is expected to contribute to in vitro neuroscience by suggesting designing principal of neural network as a reduced model of the brain.
배양된 뉴런은 실험자가 뉴런이 접하고 있는 외부환경을 바꾸는것이 용이하기 때문에, 다양한 외부환경에 따른 배양된 뉴런의 형태, 성장, 전기생리학적 활성의 변화를 관찰하는 형식으로 뇌 연구를 위한 모델로서 이용된다. 그 중 대표적인 외부환경 변화는 배양된 뉴런과 맞닿아있는 배양기판 표면에 변화를 주는 것이다. 본 학위논문은 신경 네트워크의 구조를 제어하기 위해 소프트 리소그래피 기술을 사용하고자 한다. 먼저 미세접촉 프린팅 기술을 이용해 세포 배양 기판 표면에 생체분자를 패터닝하고 그에대한 신경줄기세포의 분화와 이동성 차이를 이용해 신경 네트워크의 구조를 제어하고자 한다. 그리고 신경네트웍을 디자인 하기 위해 미세몰딩 기술을 이용해 세포 반발성 물질인 아가로즈 수화젤과 알긴산 수화젤을 미세전극칩 위에 패터닝 하고자 한다. 면역 염색법을 이용해 미세 패터닝 기술로 형성된 신경 네트워크의 생리학적 특성을 검증하고, 미세전극칩 시스템을 이용해 디자인된 신경 네트워크로부터 전기적 신호를 측정하고 전기생기학적 특성과 연결도를 분석하고자 한다.
