[不負責任研究] Proposal 進化史(一)

近幾年申請工作或是學校，Proposal這種用來展現自己的英文、研究方向、組織與邏輯、資料與情報蒐集能力的資料絕對是必備的。
2009年還是有點熟又不太熟的時候….秉持著咱們在細胞與組織工程的研究可是個未知的領域~~~Everything is possible!! 因此有了以下這份PhD研究計畫書。

說來汗顏，2013年重新申請學校時，這份2009計畫書姐可是只看了幾分鐘就直接丟進垃圾桶滴。真不知當年那些大師們在看這份計畫書的時候是否也是比照辦理呢?

這邊就別說之前不負責任研究院的那些被評為品質以上的計畫書了….現在看到那些我都很想死(掩面)  不過比起其他同事寫的… 恩，不予置評。

這邊先貼來給自己一個記錄吧!!! 諸位若是在某些地方看到類似的研究…說真的他們願意試試看且能真的作下去，也是奇葩的一種…這邊我就不追究了(遠目)。

你，也聞到了飄出來的濃濃菜味了嗎? (掩面) 這份計畫書姐花了約二個星期完成資料蒐集，paper閱讀、構思與撰寫，但當初花了另外兩個星期改英文。
超短的東西還需要兩周….就知道當年姐的英文多廢了。

Study plan
(for 2009 PhD Application)

 The research of liver bioreactor is one of the fields that I can exert my research experience of mechanical engineering, biomedical engineering, opto-electronics, system design, programming, and cell biology. This plan is a dream project about artificial organ that I wish to study in the PhD program; however, my research interest is not only limited to this realm.

Background:

The design of liver bioreactor is still a challenge because the key component for its feasibility, pure primary human hepatocytes, cannot be successful cultivated in appropriate quantities for full function [1, 2]. Many methods and hypotheses are provided to find the mechanism to solve this problem. In 2007, Hui [3] provided a micromachine silicon substrate with moving parts to demonstrate that mechanical control of tissue composition and spatial organization can facilitate the future precise investigation of the cell-cell interactions, which could be one of the key methods to solve the mechanism of the proliferation of liver cell. From Wu and Su’s research [5-7], experiments of different types of mechanical reaction, including transient contact of hepatocytes and stem cells provided by Hui [4] may also give the direction to find the relationship between force stimulate and cell proliferation. In this case, micro-fluid channels by BioMEMS devices are capable to fine-tune the flow pattern to apply on the research of this realm. For the further research on molecular and cellular of liver cell, a test of shear stress on cell differentiation and the design of complex flow patterns in bioreactor are also necessary.

Objectives:

Biomedical system design:

Design a liver bioreactor that can real time monitor and analyze the viability of human hepatocytes, and maintain available quantities of liver cell for full function. The design should consider the interaction of normal and shear stress, compression and 3D controllable turbulence by the dynamic cultivated of the hepatocyte. In addition, the observation should employ types of immunofluorescent method to detect the viability and activation of target cells. A compact motion control system is also necessary to shift the cultivated specimen of nonhuman stem cell to prevent the possible infection from different species. The real time optical monitoring system which is responsible for recording, detecting and analyzing fluorescence which ought to be low damage to all the cells in this bioreactor. Biomechanical Model built by data acquired from this system can also use for the feedback control unit on hepatocyte co-culturing real-time regulation.

Applications:

        This system can apply to determine the role of mechanical force and soluble factors in the initiation of hepatocyte proliferation. We can record and analyze cell-cell force interaction by the tunable motion process of this system.

Method (The concept of the design):

        Each unit of the bioreactor (the left of the picture) contains are show as below. Using a dish-like system (the center of the picture) with micro stepping motor (i.e Piezo electric motor) to control the position of moving parts which can regulate the interval of cell-cell contact (shift action), the gap between different cell (shift action), the exchange of the different cell types (rotate and shift), and the action of isolated regulation. The moving parts might adopt the protocol introduced by Hui [4]. By testing different parameters of gap between cells, we can find the optimal setting of liver cell proliferation. In the isolated system, the movable mechanism should responsible for removing the second specimen to a nearby region of the same dish where with independent in/out medium supply to prevent contamination. By utilizing controllable gates, we can dynamically separate those sections (the right of the picture).

For real time investigation, using LabVIEW to program the motion control system and to record the image data from the optical system integrated in the motion system that can continuously detect fluorescent signal exiting by laser plugged near the unit (does not show in the picture). In this case, mapping different types of fluorescents can find out the influence of complex bioenvironmental control factors and determine the parameters of a perfect environment to available quantities.

Reference：

1.         Andersson, H. and A. van den Berg, Microfabrication and microfluidics for tissue engineering: state of the art and future opportunities. Lab Chip, 2004. 4(2): p. 98-103.

2.         Fiegel, H.C., et al., Hepatic tissue engineering: from transplantation to customized cell-based liver directed therapies from the laboratory. J Cell Mol Med, 2008. 12(1): p. 56-66.

3.         Hui, E.E. and S.N. Bhatia, Micromechanical control of cell-cell interactions. Proc Natl Acad Sci U S A, 2007. 104(14): p. 5722-6.

4.         Hui, E.E. and S.N. Bhatia, Silicon microchips for manipulating cell-cell interaction. J Vis Exp, 2007(7): p. 268.

5.         Wu, C.C., Li, Y.S., Haga, J.H., Kaunas, R., Chiu, J.J., Su, F.C., Usami, S., and Chien, S., Directional shear flow and Rho activation prevent the endothelial cell apoptosis induced by micropatterned anisotropic geometry. Proc Natl Acad Sci U S A, 2007. 104:p. 1254-1259.

6.         Wu, C.C., Chao, Y.C., Chen, C.N., Chien, S., Chen, Y.C., Chien, C.C., Chiu, J.J., and Yen, B.L., Synergism of biochemical and mechanical stimuli in the differentiation of human placenta-derived multipotent cells into endothelial cells. J Biomech., 2008. 41 813-21.

7.         Su, F.C., Wu, C.C. and Chien, S., Microenvironment and external force in cell and tissue remodeling. J Mech Behav Biomed Mater (In Press, Invited Review)