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細(xì)胞擴(kuò)增生物反應(yīng)器控制系統(tǒng)的研究

發(fā)布時(shí)間:2018-04-28 23:04

  本文選題:波浪式生物反應(yīng)器 + 控制系統(tǒng); 參考:《中國(guó)人民解放軍軍事醫(yī)學(xué)科學(xué)院》2017年碩士論文


【摘要】:目的:近年來(lái),隨著科學(xué)技術(shù)的發(fā)展,動(dòng)物細(xì)胞培養(yǎng)技術(shù)被廣泛應(yīng)用于生物醫(yī)藥產(chǎn)品的工業(yè)生產(chǎn)中,如蛋白質(zhì)藥物研發(fā)、干細(xì)胞移植、疫苗生產(chǎn)、人造組織器官等領(lǐng)域。例如造血干細(xì)胞移植可長(zhǎng)期重建造血和免疫,它適用于治療造血干、祖細(xì)胞或相關(guān)基因有缺陷的疾病,如白血病、重度免疫缺損、自身免疫病等,是一種重要的生物治療或細(xì)胞治療方法。但往往人體本身可提供的造血干細(xì)胞不足,這就迫切需要在體外對(duì)這些干細(xì)胞進(jìn)行大規(guī)模擴(kuò)增。生物反應(yīng)器的提出就為造血干細(xì)胞的體外擴(kuò)增提供了一種非常有效的方法。細(xì)胞的培養(yǎng)擴(kuò)增過(guò)程是極其復(fù)雜的生物化學(xué)反應(yīng)過(guò)程,其代謝必須在適宜的周?chē)h(huán)境中才能有效進(jìn)行。國(guó)外已經(jīng)有比較成熟的生物反應(yīng)器,但國(guó)內(nèi)還沒(méi)有商品化的動(dòng)物細(xì)胞生物反應(yīng)器。本文對(duì)波浪式生物反應(yīng)器的控制系統(tǒng)進(jìn)行研究,旨在完成一套基于波浪式生物反應(yīng)器的細(xì)胞培養(yǎng)條件的控制系統(tǒng),使其能更好的應(yīng)用在細(xì)胞的擴(kuò)增培養(yǎng)上,進(jìn)而推動(dòng)我國(guó)生物反應(yīng)器行業(yè)向前發(fā)展。方法:通過(guò)對(duì)當(dāng)前生物反應(yīng)器培養(yǎng)條件的控制方法和控制系統(tǒng)進(jìn)行分析和討論,提出適合波浪式生物反應(yīng)器培養(yǎng)系統(tǒng)的控制方法。控制系統(tǒng)的設(shè)計(jì)主要可以分為三部分:培養(yǎng)條件控制方法的設(shè)計(jì)、控制系統(tǒng)軟/硬件部分設(shè)計(jì)、仿真及實(shí)際實(shí)驗(yàn)驗(yàn)證部分。關(guān)于控制方法的設(shè)計(jì),根據(jù)溫度控制要求及溫度控制大滯后的特點(diǎn),設(shè)計(jì)出Fuzzy-PID控制算法;根據(jù)PH控制要求及PH過(guò)程強(qiáng)烈非線性的特性,對(duì)分段式變?cè)鲆鍼ID進(jìn)行改進(jìn),設(shè)計(jì)出四區(qū)段變?cè)鲆鍼ID控制算法;根據(jù)溶解氧濃度非定值控制的特點(diǎn),設(shè)計(jì)出TP-PID的控制算法。關(guān)于控制系統(tǒng)軟/硬件部分的設(shè)計(jì),采用單片機(jī)為主控芯片,結(jié)合MPLAB、MATLAB等實(shí)現(xiàn)上、下位機(jī)的連接與配合。實(shí)驗(yàn)部分,在仿真實(shí)驗(yàn)的基礎(chǔ)上進(jìn)一步通過(guò)實(shí)際實(shí)驗(yàn)驗(yàn)證控制算法的效果以及控制系統(tǒng)的性能。內(nèi)容:本文研究工作主要包括以下幾個(gè)方面:(1)生物反應(yīng)器控制系統(tǒng)分類(lèi)及參數(shù)控制方法調(diào)研、控制系統(tǒng)控制方案設(shè)計(jì)。通過(guò)大量查閱文獻(xiàn)并與相關(guān)行業(yè)人士接觸,調(diào)研生物反應(yīng)器細(xì)胞培養(yǎng)的最新進(jìn)展情況。分析各類(lèi)控制方法和控制系統(tǒng)的優(yōu)缺點(diǎn),從而為文章控制系統(tǒng)的選擇和控制方法的提出提供研發(fā)的現(xiàn)實(shí)意義與應(yīng)用前景。設(shè)計(jì)系統(tǒng)整體控制方案,通過(guò)比較硬件元器件的性能以及滿足應(yīng)用的情況,選出最佳的系統(tǒng)方案。(2)控制系統(tǒng)硬件設(shè)計(jì)與實(shí)現(xiàn)。系統(tǒng)硬件設(shè)計(jì)主要包括下位機(jī)執(zhí)行部件的選擇、上位機(jī)主控部件的選擇以及上、下位機(jī)之間的集成。下位機(jī)硬件部分由主控芯片、檢測(cè)器件、執(zhí)行器件、傳輸線等幾部分組成。主控芯片選擇dspic30f6014a單片機(jī)為控制系統(tǒng)下位機(jī)的核心,該芯片將核心處理層及整個(gè)外圍電路層如輸入/輸出端口、內(nèi)存、定時(shí)器、計(jì)數(shù)器等全部都集成在一塊芯片上,實(shí)現(xiàn)了實(shí)驗(yàn)數(shù)據(jù)的一整套的接收、計(jì)算、存儲(chǔ)、發(fā)送功能;檢測(cè)器件根據(jù)需要控制的T、PH、DO三個(gè)培養(yǎng)條件分別選擇Pt100、在線PH儀、在線溶解氧儀;執(zhí)行器件根據(jù)控制原理選擇電熱毯、蠕動(dòng)泵、電磁閥等。上位機(jī)主控部件根據(jù)需要選擇集顯示、控制、存儲(chǔ)等功能于一體的DGUS屏,實(shí)現(xiàn)對(duì)控制系統(tǒng)的整體監(jiān)控。上、下位機(jī)集成主要由它們之間的通訊線連接及統(tǒng)一的通訊協(xié)議實(shí)現(xiàn)。(3)控制系統(tǒng)控制算法設(shè)計(jì)與實(shí)現(xiàn)。本文主要設(shè)計(jì)完成了三個(gè)控制算法,即針對(duì)溫度控制的Fuzzy-PID控制算法、針對(duì)PH控制的四區(qū)段變?cè)鲆鍼ID控制算法和針對(duì)溶解氧濃度控制的TP-PID控制算法。其中,Fuzzy-PID控制算法并非采用傳統(tǒng)的并聯(lián)使用的模式,而是將兩種方法整合到一起,以檢測(cè)信號(hào)作為模糊控制的輸入,以模糊控制的輸出作為PID控制的輸入,最后以PID控制的輸出作為系統(tǒng)控制信號(hào)的輸出;四區(qū)段變?cè)鲆鍼ID控制是在分段式變?cè)鲆鍼ID的基礎(chǔ)上根據(jù)生物反應(yīng)器的實(shí)際情況進(jìn)行改進(jìn)得到的,充分考慮了PH控制的強(qiáng)烈的非線性特性;溶解氧濃度控制算法的設(shè)計(jì)考慮只需將其控制在某一范圍內(nèi)即可,主要是參考傳統(tǒng)控制的模式。(4)人機(jī)交互界面設(shè)計(jì)及觸控配置完成。上位機(jī)人機(jī)交互界面選用DGUS屏,型號(hào)為DWT80600T080_06WT,使用Microsoft Visio Premium 2010進(jìn)行圖片制作,需要顯示的圖片主要包括系統(tǒng)初始化部分、主界面部分、數(shù)據(jù)輸入界面部分等。觸控配置使用DGUS配置工具V49,將制作好的圖片導(dǎo)入配置工具,按照顯示屏操作要求在相應(yīng)區(qū)域位置添加文本顯示、按鍵返回、RTC顯示等配置操作,并設(shè)置好相應(yīng)配置的變量地址、文本長(zhǎng)度、按鍵值、案件效果等。將配置好觸控功能的顯示圖片導(dǎo)入到DGUS屏里就完成了人機(jī)交互界面的設(shè)計(jì)。(5)仿真實(shí)驗(yàn)加實(shí)際實(shí)驗(yàn)驗(yàn)證算法可靠性及控制系統(tǒng)性能。為驗(yàn)證控制算法的可靠性,首先使用MATLAB軟件中的Simulink軟件包設(shè)計(jì)Fuzzy-PID的溫度控制仿真實(shí)驗(yàn)和分段式變?cè)鲆鍼ID的PH控制仿真實(shí)驗(yàn),兩個(gè)仿真實(shí)驗(yàn)均以控制時(shí)間和控制精度作為控制算法性能可靠的判斷依據(jù)。仿真實(shí)驗(yàn)驗(yàn)證算法可靠性之后設(shè)計(jì)實(shí)際實(shí)驗(yàn)進(jìn)行驗(yàn)證。使用MPLAB軟件按照算法設(shè)計(jì)編寫(xiě)三種控制算法的程序并將其導(dǎo)入單片機(jī)進(jìn)行實(shí)際實(shí)驗(yàn),驗(yàn)證標(biāo)準(zhǔn)依然是控制的時(shí)間和穩(wěn)定后的控制精度。結(jié)果:本文根據(jù)細(xì)胞培養(yǎng)的要求,完成了波浪式生物反應(yīng)器控制系統(tǒng)中T、PH、DO三個(gè)培養(yǎng)條件控制算法的設(shè)計(jì),完成了控制系統(tǒng)硬件選擇,完成了控制系統(tǒng)電路設(shè)計(jì),完成了上、下位機(jī)通訊設(shè)計(jì),完成了人機(jī)交互界面的選擇和設(shè)計(jì),完成了單片機(jī)對(duì)應(yīng)程序的編寫(xiě),最后在仿真實(shí)驗(yàn)基礎(chǔ)上設(shè)計(jì)實(shí)際實(shí)驗(yàn)完成了算法可靠性的驗(yàn)證工作。發(fā)表了兩篇論文。結(jié)論:本文介紹了各類(lèi)生物反應(yīng)器控制系統(tǒng)及控制方法,具體分析了各控制系統(tǒng)的優(yōu)缺點(diǎn),提出采用單片機(jī)作為主控芯片對(duì)波浪式生物反應(yīng)器控制系統(tǒng)進(jìn)行研究,并根據(jù)要調(diào)節(jié)的培養(yǎng)條件的特性設(shè)計(jì)出不同的控制方法。在實(shí)際實(shí)驗(yàn)進(jìn)行控制時(shí),溫度控制精度可達(dá)到±0.1℃,PH控制精度可達(dá)到±0.05,溶解氧濃度精度可達(dá)到±6%。表明本文所研制的控制系統(tǒng)能夠穩(wěn)定可靠的運(yùn)行且控制效果良好,說(shuō)明控制系統(tǒng)能很好地維持細(xì)胞生長(zhǎng)需要的適宜的環(huán)境,滿足細(xì)胞培養(yǎng)過(guò)程的控制要求。本文的創(chuàng)新點(diǎn)在于根據(jù)不同被控條件設(shè)計(jì)出相應(yīng)的控制方法,如針對(duì)溫度控設(shè)計(jì)了模糊PID控制、針對(duì)PH控制設(shè)計(jì)了四區(qū)段變?cè)鲆鍼ID控制;使用dsPIC30f6014a型號(hào)單片機(jī)作為主控芯片,不僅降低了研發(fā)成本,還提高了系統(tǒng)靈活性,縮短了開(kāi)發(fā)周期;使用DGUS屏作為人機(jī)交互界面,實(shí)現(xiàn)了對(duì)控制界面的搭配式設(shè)計(jì);將系統(tǒng)進(jìn)行模塊化設(shè)計(jì),最后集合到一起,降低了故障的影響率。
[Abstract]:Objective: in recent years, with the development of science and technology, animal cell culture technology has been widely used in the industrial production of biological medicine products, such as protein drug development, stem cell transplantation, vaccine production, artificial tissue and organs, such as hematopoietic stem cell transplantation for long period of heavy construction of blood and immunity, it is suitable for the treatment of hematopoietic stem, ancestral fine. Diseases such as leukemia, severe immunodeficiency, and autoimmune diseases, such as leukemia, severe immunodeficiency, and autoimmune diseases, are an important method of biological treatment or cell therapy. However, the human body itself can provide a lack of hematopoietic stem cells. This is an urgent need for large-scale expansion of these stem cells in vitro. The bioreactor is proposed for hematopoiesis. The expansion of stem cells in vitro provides a very effective method. The process of cell culture and amplification is an extremely complex biochemical reaction process, and its metabolism must be effectively carried out in a suitable environment. There have been more mature bioreactors abroad, but there are no commercialized animal cell bioreactors at home. The paper studies the control system of wave bioreactor, which aims to complete a set of control system based on cell culture conditions based on wave bioreactor, so that it can be better applied to cell expansion and culture, and then promote the development of bioreactor industry in China. The control method and control system are analyzed and discussed, and the control method suitable for the wave bioreactor training system is put forward. The design of the control system can be divided into three parts: the design of the training condition control method, the design of the soft / hardware part of the control system, the simulation and the actual experimental verification part. The Fuzzy-PID control algorithm is designed for the requirements of temperature control and the large lag in temperature control. According to the requirements of PH control and the strong nonlinear characteristics of the PH process, the piecewise variable gain PID is improved and the four section variable gain PID control algorithm is designed. The control algorithm of the TP-PID is designed according to the characteristics of the dissolved oxygen concentration control. In the design of the soft / hardware part of the control system, the MCU is used as the main control chip, the connection and coordination of the lower computer are combined with MPLAB, MATLAB and so on. In the experiment part, the effect of the control algorithm and the performance of the control system are verified on the basis of the simulation experiment. The following aspects: (1) the research on the classification of the bioreactor control system and the method of parameter control, the design of control system control scheme. Through a large number of references and contact with the related industry, the latest progress in the cell culture of bioreactor is investigated and the advantages and disadvantages of various control methods and control systems are analyzed, so as to control the system The selection and control methods provide the practical significance and application prospect of the research and development. Design the overall control scheme of the system. By comparing the performance of the hardware components and meeting the application situation, the best system scheme is selected. (2) the hardware design and implementation of the control system. The system hardware design mainly includes the selection of the executive components of the lower machine. The hardware part of the lower computer consists of the main control chip, the detection device, the execution device, the transmission line and so on. The main control chip chooses the dspic30f6014a MCU as the core of the control system lower computer. The chip will be the core processing layer and the whole peripheral circuit layer, such as input / output. The mouth, memory, timer, counter and so on are all integrated on a chip, realizing a complete set of receiving, calculating, storing and sending function of experimental data. The detection device selects Pt100, online PH instrument, on-line dissolved oxygen system according to the three conditions of T, PH, DO, and the actuator chooses electric blanket and peristaltic pump according to the control principle. The main control components of the host computer, according to the needs, select the DGUS screen which integrates display, control, storage and so on. The control system is monitored as a whole. The integration of the lower computer is mainly connected by the communication line between them and the unified communication protocol. (3) the design and implementation of control system control algorithm. The main design and completion of this paper is to complete the design and implementation of the control system. Three control algorithms, namely, the Fuzzy-PID control algorithm for temperature control, the four section variable gain PID control algorithm for the PH control and the TP-PID control algorithm for the dissolved oxygen concentration control, the Fuzzy-PID control algorithm is not used in the traditional parallel mode, but the two methods are integrated together to detect the signal as a signal. The input of fuzzy control takes the output of fuzzy control as the input of PID control, and the output of PID controlled as the output of the system control signal, and the four section variable gain PID control is improved on the basis of the piecewise variable gain PID based on the actual situation of the bioreactor, taking full account of the strong non line of the PH control. The design of dissolved oxygen concentration control algorithm only needs to control it in a certain range, mainly reference to the traditional control mode. (4) human-computer interaction interface design and touch control configuration. The computer interaction interface of the upper computer is DGUS screen, the model is DWT80600T080_06WT, and the Microsoft Visio Premium 2010 is used for the picture system The pictures that need to be displayed mainly include the system initialization, the main interface part, the data input interface, etc. the touch control configuration uses the DGUS configuration tool V49 to import the good picture into the configuration tool, and add the text display, the key return, the RTC display and so on in accordance with the display operation requirements. The corresponding variable address, text length, key value, case effect, etc.. The design of human-computer interaction interface is completed by introducing the display pictures with good touch function into the DGUS screen. (5) simulation experiments and practical experiments verify the reliability of the algorithm and the performance of the control system. In order to verify the reliability of the control algorithm, the first use of the MATLAB software is to verify the reliability of the algorithm. The Simulink software package designs the temperature control simulation experiment of Fuzzy-PID and the PH control simulation experiment of the piecewise variable gain PID. The two simulation experiments all take the control time and the control precision as the reliable judgment basis of the control algorithm performance. The simulation experiment verifies the reliability of the algorithm and designs the actual experiment to verify. The MPLAB software is used in the simulation experiment. According to the algorithm, three kinds of control algorithms are programmed and introduced into the single chip computer to carry out the actual experiment. It is proved that the standard is still the control time and the control precision after the stability. According to the requirements of cell culture, this paper completed the design of three training conditions control algorithms of T, PH and DO in the wave bioreactor control system. The control system hardware selection, completed the control system circuit design, completed the upper and lower computer communication design, completed the selection and design of the human-computer interaction interface, completed the programming of the single chip computer corresponding program. Finally, on the basis of the simulation experiment, the actual experiment was designed to complete the verification of the reliability of the calculation method. Two papers were published. In this paper, the control system and control methods of various bioreactors are introduced, the advantages and disadvantages of each control system are analyzed, and the control system of wave bioreactor is studied by using single chip microcomputer as the main control chip, and different control methods are designed according to the characteristics of the training conditions to be adjusted. The precision of temperature control can reach to 0.1 C, the precision of PH control can reach 0.05, the precision of dissolved oxygen concentration can reach to + 6%., which indicates that the control system developed in this paper can be stable and reliable, and the control effect is good. It shows that the control system can maintain the suitable environment for cell growth well and meet the control of cell culture process. The innovation point of this paper is to design the corresponding control methods according to the different controlled conditions, such as the design of the fuzzy PID control for temperature control, the design of the four section variable gain PID control for the PH control, and the use of the dsPIC30f6014a single chip as the main control chip, which not only reduces the R & D cost, but also improves the system flexibility and shortens the opening. With the use of DGUS screen as a human-computer interface, the collocation design of the control interface is realized. The system is designed and finally assembled to reduce the impact rate of the fault.

【學(xué)位授予單位】:中國(guó)人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2017
【分類(lèi)號(hào)】:TP273;Q813

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