| With the development and success of space technology and flight, our country has enter the stage of space researches and explorations. Since the complexity of space environment, the instruments for space loading experiments are restricted for some special conditions. However, the biological experimental methods on ground are lack of proper space loading instruments which means that they cannot play a role in space. The researches on explorations of small volume, low power consumptive and automated instruments are under developing with the support of many national key scientific projects, like 973, 863, National Sci-Tech Support Plan, etc. Microfluidic chips with the functions of microscale size, integration and high throughput, broken the restriction of traditional biology, are ideal methods for space loading experiments. The controllable fluid and versatile micro-design can help bulding similar models of space biology. Related researches have indicated that there might be changes of neural and immune functions under radiations and microgravity. To build up neural and immune cell culture modes in microfluidic chip can help us understand the mechanism of space biological effect. What’s more, by creating the culture modes, they can play an important role in broden biological research area, like drug delivery, cell analysis and cell-cell interaction, etc.This paper relied on microfluidic chip technique to build up neural and immune cell culture states. At the same time, a whole microfluidic chip device and and a pneumatic driven system were built up for both service for cell culture. The following key technologies were founded:(1) To build up multi-drug stimulate experiments in microfluidic chip based on neural cell culture;(2) Fabricate a microfluidic chip which can maintain long tern perfusion culture of immune suspended cell and explore the cell states in chip;(3) A fabrication method of microfluidic chip was proposed and established multi-mammal adherent cell culture using the fabricated chip;(4) Invent a miniaturized instrument contained temperature control, pumping driven and on-site detection which meet the demanding of space loading experiments;(5)With the development of pneumatic microvalves chip and related driven system, multi-cell culture and co-culture modes were presented. The research content mainly includes the following aspects:1) Design and fabricate free-template microfluidic chips by UV-cured glue(NOA 81), achieved various cell culture conditions on the chips. The proposed method is more suitable for the early structure exploration stage of microfluidic chip than existing procedures, since no templates are needed. Fluid flow can be controlled effectively and contamination can be resisted by simple PDMS-NOA 81 linked microvalves embedded in the microfluidic chip. SH-SY5 Y, U87, HCT 116, Hela and Hek 293 t were integrated into the chip and maintain normal viabilities up to one week with stable perfusion. Concentration gradient was generated to investigate the SH-SY5 Y cells in the microfluidic chip responded to DA drug stimulation. The cells appeared different in terms of shape and proliferation which strongly relied on drug concentration. The outcomes demonstrated the potential applications of our biocompatible NOA 81 chip in space and other various cell analyses.2) A microfluidic chip was proposed which can entrap cells and avoid fluid stress efficiently. Immune cell can maintain long tern culture in the chip with perfuson. After the exploration of material and cell line, NOA 81 chips were first adopt to study the proper chip design. And the outcome indicated one of the key elements was effective rejection design. Subsequently, PDMS chip provide the decisive influence of the flow rate which can damage the cell by shear stress. Finally, polyvinyl chloride fabricated microfluidic chip combined bolting-silk was proposed and maintained long-tern suspended cell culture, which may give strong support for the study of cell culture in space environment.3) A whole microfluidic cell culture device containing accurate temperature control, adjustable pumping system and on-site detection was proposed for the application of space research. Temperature controlling system was combined MSP 430 microcontroller with PID algorithm and maintained the cell culture chamber at 37°C ± 0.5°C. Two pumping system were studied to observe the fluid flow influence to cell culture. Imaging device can upload the real-time observation video to the remote sensing platform and ensure the detected of cell growth, proliferation and movement in space environment. By using SH-SY5 Y cell as an example, different materials anddesigns of microfluidic chip were adopted to observe their affect to cell culture. The small volume, low power consumptive and automated instruments with real-time detection may perfectly meet the demanding of space loading experiment for cell culture.4) A multi-functional pneumatic microvalve chip with related driven system was proposed here. Standard template chip design and exterior gas control device were presented in detail. Dye experiments prove the possibilities of three independent chambers that can be either linked or blocked by microvalves with the cooperation of the exterior gas controlling device. In order to increase the cells viabilities in chip, different treatments were also tested on PDMS substrates. Different cell line-SH-SHSY, Hela and HTC-116 were vaccinated respectively and cultured for 96 h. Three transfected SH-SY5 Y cells were also seeded and treated with MPP+ to observe the assembling of α-syn. SH-SY5 Y cells co-cultured with SH-SY5 Y, HCT11 and Hela were induced into three independent chambers to observe the neural protection and interaction. The results revealed the practicability of our entire multi-functional pneumatic chip system and also showed significant potential abilities in disease diagnosis, drug delivery, clinical test and many other research areas.Neural and immune cell culture conditions were both established in microfluidic chip. A miniaturized microfluidic cell culture device was also build up for the study of space life science and space loading mission. At the same time, the invention of multi-functional pneumatic microvalve chip with related driven system fully broaden the potential applications in various research areas. |
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