200825168 九、發明說明： 【發明所屬之技術領域】 本發明係有關於一種微重力I_ 刀無囷裱境細胞自動培養 裝置，旨在提供一種具微重力益结⑽ 刀無囷％境及可自動更換 基之細胞培養裝置。 【先前技術】200825168 IX. INSTRUCTIONS: [Technical field of invention] The present invention relates to a microgravity I_ knife-free environment cell automatic culture device, which aims to provide a microgravity benefit knot (10) knife without flaws and can be automatically Replace the base cell culture device. [Prior Art]
由於細胞體外培養需靠外在環境控制⑽、〇2、鱼菌、 溫度）及養分供給，錄需、_天至朗才能培養出所需 之細胞數量，對於養份供給及“純·或設備交互使 用’，理過程容易使細胞受到傷害影響增生，且如果係經 由人員來操作，也不易精確定量。 然而，生物技術發展迅速且細胞需求量增加， 程及生技藥物開發已成為發展趨勢。雖有生物哭產。 但細胞增生速度仍無法達到需求(三天約:產二 =)m達到節省人力之功能。爰此可藉著 ,技術將多種設儳整合為— =正 供給，因此使自動化及藥劑適時定量 減少。當不利^ 響細胞增生因素得以大幅 培養。 、讀增生條件減少，即可將細胞快迷大量 方式可 分二計上有許多不同的類型’培養 CD平面式：將細胞放置於特定區域，培養基以漭動 6 200825168 方式或定時更換作為培養系統。 (2)立體式：將細胞置於支架上，再將整個支架置於 培養液中，培養基採定時更換方式。Because the cell culture in vitro requires external environment control (10), 〇2, fish bacterium, temperature) and nutrient supply, the demand for the _ Tianzhilang can cultivate the required number of cells, for nutrient supply and "pure or device interaction Using ', the process is easy to cause cells to be affected by damage, and if it is operated by personnel, it is not easy to accurately quantify. However, biotechnology is developing rapidly and cell demand is increasing, and the development of biotechnology and drug has become a trend. Crying, but the rate of cell proliferation still can not meet the demand (three days about: production two =) m to achieve the function of saving manpower. This can be achieved by technology, the integration of a variety of settings into - = positive supply, thus making automation and pharmaceuticals Timely quantitative reduction. When the unfavorable cell proliferation factor is greatly cultivated, and the proliferative condition is reduced, the cell can be fascinated in a large number of ways. There are many different types of 'culture CD plane: placing cells in specific areas The medium is used as a culture system by using the method of swaying 6 200825168 or timing. (2) Stereoscopic: Place the cells on the stand and then The entire scaffold is placed in the culture medium and the medium is replaced at regular intervals.
與平面培養皿（Gultur*e_dish)料料比較後，平面 式培養方法中細胞增生效率並非十分快速，且設備上並未 大幅減少’社體式培制受限於支”造方法，細胞無 法很均勻地放置於支架當巾，且支架特性不—細胞無法 於立體方向上生長，因此有厚度限制。 美國太空總署研究資料指出，利用平面培養皿進行細 胞培養時由於受到地心引力仙，細胞只能往橫方向掩 生’因此細胞增生速度緩慢，並經實驗驗證在太空中^ 騎境下，細射在衫地列力之翻，自由地往各個 方向增生，可達到細胞增生的效果。 近年來雖有研究學者利㈣統培養箱驗證美國太* =署微：力理論增生細胞之效果’但上述操作仍; 業，氣體交換使用㈣方式⑽、ϋ2)，而且—次 乍 ==需重新設定氣體濃度，不但費時費力也不能直 抑達到貝用證明，目前仍沒有一套整合自動化作業、無 環境控制及微重力細胞增生機制於—導: 增生速度仍然受到限制。11 ¥致, 另外i統培養室之裝備如第十四圖所示，有水地 室(A5)晾乾架(A2)、消毒器(A3)、無菌操作室(A4)、準俺 、電冰箱⑽）、離心機⑽、恆溫培養箱⑽、^ 200825168 微鏡天平台（A9) '實驗台（Αίο)、污物桶（A11)、藥品櫃 (A12)、器械櫃（A13)等，其裝備相當多，無法集中，故所 需的場地相當大，操作時需於整個室内活動，相當不方便。 又，傳統之培養盤内之培養基係以人工手動置換，於 更換過程容易毁傷細胞，或導致細胞壞死。 【發明内容】 爰此，本發明係以解決習用以人工手動培養細胞容易 使細胞壞死及習用之細胞培養所需的場地過大等問題。 > 本發明提供一種微重力無菌環境細胞自動培養裝 置，其係設置主架體，而主架體内係設有培養箱、、培養基 運运模組、電控模組、無菌操作室、去離子水運送模組、 二氣化碳濃度控制模組、氣壓控制箱及廢液儲存桶，其中·· _培養箱，包含有微重力控制模組及恆溫模組，而微重力控 1模組係包括有微重力控制單元、培養基換取機構'管路 清洗模組，另，培養基換取機構設有第一幫浦、第一管路， •且第一官路接合有三通閥，而管路清洗模組設有第二管路 f去離子水儲存桶，第二管路亦接合於三通閥，另，恆溫 ' ◊、係叹有加熱控制器及溫度感測器，加熱控制器係組 '又於培養箱之外圍，而溫度感測H設於培養箱内部；培養 土運送权組，係設有第三管路，第三管路係接合於三通 -=其並設有冷藏箱，以保存培養基，透過第三管路運:培 < 土至微重力控制模組之培養基換取機構。 上述之微重力控制單元設有紅外線定位器、培養盤、 8 200825168 第一快速接頭、第二快速接頭、控制盒、通氣孔及第—氣 壓缸。 上述之培養基換取機構另設有出水口、進水口、擋板 及第二氣壓缸。 上述之管路清洗模組另設有蓄液槽。 上述之培養基換取機構之第一管路加設有加熱單元。 上述之加熱單元係設有水浴加熱槽、恆溫控制器，而Compared with the flat culture dish (Gultur*e_dish) material, the cell proliferation efficiency in the planar culture method is not very fast, and the equipment is not significantly reduced in the 'corporate culture limited by the branch' method, the cells can not be very uniform Placed on the scaffold as a towel, and the characteristics of the scaffold are not - the cells can not grow in the stereo direction, so there is a thickness limit. The NASA research data indicates that when using cell culture in a flat culture dish, the cells are only affected by gravity. Can be shaded in the horizontal direction 'so the cell proliferation rate is slow, and experimentally verified in space ^ riding the environment, fine shot in the shirt and the force of the flip, free to proliferate in all directions, can achieve the effect of cell proliferation. Although there are research scholars (4) unified incubator to verify the US too * = Department of Micro: force theory of the effect of proliferating cells 'but the above operations are still; industry, gas exchange use (four) way (10), ϋ 2), and - times 乍 = = need to re Setting the gas concentration is not only time-consuming and laborious, but also can not directly achieve the proof of use. There is still no integrated automation operation, no environmental control and micro-gravity The mechanism of cell proliferation is guided by: The rate of proliferation is still limited. 11 ¥, In addition, the equipment of the i-culture room is shown in Figure 14, there is a water room (A5) drying rack (A2), sterilizer (A3) ), aseptic operation room (A4), quasi-俺, refrigerator (10), centrifuge (10), constant temperature incubator (10), ^ 200825168 micro-mirror platform (A9) 'laboratory (Αίο), dirt bucket (A11), medicine Cabinets (A12), instrument cabinets (A13), etc., are equipped with quite a lot of equipment and cannot be concentrated. Therefore, the required space is quite large, and it is necessary to operate throughout the room during operation, which is quite inconvenient. Moreover, the culture medium in the traditional culture tray Manually replacing by hand, it is easy to damage cells during the replacement process, or cause cell necrosis. SUMMARY OF THE INVENTION Accordingly, the present invention is intended to solve the problem that the field used for artificially culturing cells to easily cause cell necrosis and conventional cell culture is too large. The present invention provides a microgravity aseptic environment cell automatic culture device, which is provided with a main frame body, and the main frame body is provided with an incubator, a medium transport module, an electronic control module, and an aseptic operation. Room, deionized water transport Group, two gasification carbon concentration control module, air pressure control box and waste liquid storage tank, wherein ·· _ incubator contains microgravity control module and constant temperature module, and microgravity control 1 module system includes micro The gravity control unit and the medium exchange mechanism 'the pipeline cleaning module, and the medium exchange mechanism is provided with the first pump and the first pipeline, and the first official road is connected with the three-way valve, and the pipeline cleaning module is provided The second pipeline f is a deionized water storage tank, the second pipeline is also joined to the three-way valve, and the constant temperature '◊, the sigh has a heating controller and a temperature sensor, and the heating controller system group' is in the incubator On the periphery, the temperature sensing H is set inside the incubator; the culture soil transport right group is provided with a third pipeline, and the third pipeline is connected to the tee-= it is provided with a refrigerator to preserve the culture medium. Transported through the third pipeline: the culture medium exchange mechanism of the soil-microgravity control module. The above microgravity control unit is provided with an infrared positioner, a culture plate, 8 200825168 first quick connector, a second quick connector, a control box, a vent hole and a first air cylinder. The medium exchange mechanism described above additionally has a water outlet, a water inlet, a baffle and a second pneumatic cylinder. The pipeline cleaning module described above is further provided with a liquid storage tank. The first line of the above medium exchange mechanism is provided with a heating unit. The heating unit described above is provided with a water bath heating tank and a thermostat controller, and
水浴加熱槽内並設有葉片，且水浴加熱槽内之第三 糸 以繞行方式置入。 係 養基加熱至適合 上述之第三管路繞行之長度係使培 細胞生長之溫度。 控制單元、控制電 上述之電控模組設有電源供應器 腦0 氣化碳濃度控制模組 上述之無菌操作室室内伤#古 水槽，室外則設有攄網、抽風:。殺菌光源、水龍 度感=之錢妓料㈣包衫二氣化; 上述之三通闕之第二管 路’另’三關之第二管路為斷路I時/第三管路印名 上述之第一幫浦、第二幫浦係：：：官路即為通* 上述之廢液儲存桶 '、二蠕動式幫浦。 係以回收廢去離子水及廢格 9 200825168 基。 本發明具有下列之優點： 1. 本發明藉著控制整合技術將多種設備整合為一，透 過感測及定量元件控制培養環境，並使培養基複雜交換過 程自動化及藥劑適時定量供給，因此使得培養過程中會影 響細胞增生因素得以大幅減少。當不利與細胞增生條件減 少，即可將細胞快速大量培養。 2. 藉由本發明不論是平面式或立體式之細胞培養均 • 可適用，而且其增生附著狀態更為均勻。 3. 本發明全程自動化，可減少人為疏失及污染，有利 於細胞之培養，對於細胞之質與量皆有大幅之提昇。 1本發明將多種設備整合為一，有效節省室内空間， „ 可有效簡化流程、節省成本。 【實施方法】 本發明係為一種微重力無菌環境細胞自動培養裝 _ 置，請先參閱第一圖，本圖係為本發明主要構件說明圖〔較 細部零件圖中未示〕，其係設置主架體（1)，而主架體 (1)内係設有培養箱（2)、培養基運送模組（3)、電 控模組（4)、無菌操作室（5)、去離子水運送模組（6)、 二氣化碳濃度控制模組（7)、氣壓控制箱（8)及廢液儲 存桶（9)，其中： " 培養箱（2)，包含有微重力控制模組（21)及恆溫 模組（22);而微重力控制模組（21)係包括有微重力控 200825168 制單元（211)、培養基換取機構（212)、管路清洗模組 (213)’另，培養基換取機構（212)設有第一幫浦（2121)、 第一管路（2122)，且第一管路（2122)接合有三通閥 (2123)，而管路清洗模組（213)設有第二管路（2131)、 蓄液槽（2132)’第二管路（2131)亦接合於三通閥（2123)， 另，恆溫模組（22)，係設有加熱控制器（221 )及溫度 感測器（ 222)，加熱控制器（221)係組設於培養箱（2) 之外圍’而、azL度感測裔（ 222)設於培養箱（2)内部。 .另，管路清洗模組（213)之第二管路（2131)係組 接去主架體（1)之離子水儲存桶（n)。 並，培養基換取機構（212)之第一管路（2122)加 -設有加熱單元（214)，加熱單元（214)係設有水浴加熱 -槽（2142)、恆溫控制器（2141 )，而水浴加熱槽（2142) 内並設有葉片（2143)，且水浴加熱槽（2142)内之第一 官路（2122)係以繞行方式置入，第一管路（2122)繞行 _之長度係使培養基加熱至適合細胞生長之溫度。 另，微重力控制單元（211 )設有紅外線定位器 (2111)、培養盤（2112)、第一快速接頭（2113)、第 第一快速接頭（2114)、控制盒（2115)、通氣孔（2116) 及第一氣壓缸（2117)。 而培養基換取機構（212)另設有出水口（2124)、 進水口（2125)、擋板（2126)及第二氣壓缸（2127); 復，微重力控制單元（211)固定於一基板（23)上，基 11 200825168 板（23)底部設有滑槽（24)，將基板（23)之滑槽（24) 組合於培養箱（2)所設之滑軌（25)，使兩者可相對滑 移。 培養基運送模組（3)，係設有第三管路（31)，第 三管路（31)係接合於三通閥（2123)，並設有冷藏箱（32)， 以保存培養基、培養試劑及細胞生長因子；透過第三管路 (31)運送培養基至微重力控制模組（21)之培養基換取 機構（212)。 . 電控模組（4)設有電源供應器（41)、控制單元（42)、 控制電腦（43)，而電控模組（4)係以人為操作控制電 腦（43)，控制電腦（43)再以控制單元（42)監控恒溫 ‘模組（22)、二氣化碳濃度控制模組（7)，另，電源供 -應器（41)係以供應所有需要電源之組件。 無菌操作室（5)係設有UV殺菌光源（51)、水龍頭 (52)、水槽（53)、濾網（54)、抽風機（55)，該水 φ槽（Μ)不使用日守可以蓋板〔圖中未示〕覆蓋，以加大無 菌操作室（5)内可使用之平面空間；該抽風機（55)之 作用係在於無菌操作室（5)之門若打開時，抽風機（55) 則會立刻將外部空氣經由濾網（54)抽入無菌操作室 至内，再由UV殺菌光源（51)消毒抽入之外部空氣，夢 以使無菌操作室（5)維持無菌空間，並使無菌操作室（5曰) 形成一正壓空間。 去離子水運送模組（6)，係設有第二幫浦（61)、 12 200825168 第四管路（62) ’由去離子水館存桶（11)運送去離子水 至無菌操作室（5)之水龍頭（52)。 二氣化碳濃度控制模組（Ό，係包括有二氣化碳濃 度感測器（71)、電磁閥（72)。 氣壓控制箱（8)，係以控制微重力控制單元（211) 之第一氣壓缸（2117)及培養基換取機構（212)之第二 氣壓缸（2127)。 廢液儲存桶（9)，係以回收廢去離子水及廢培養基。 • 如第二圖所示，係為主架體（1)及上述各部份組件 之配置圖，於圖中可見： 主架體（1)。 - 培養箱（2)。 電控模組（4)之電源供應器（41)、控制單元（42)、 控制電腦（43) 無菌操作室（5)室内係設有UV殺菌光源（51)、水 鲁龍頭（52)、水槽（53)，室外則設有濾網（54)、抽風 機（55) 〇 加熱單元（214)〔加設於培養基運送模組（3)之第 三管路（31)〕，係設有恒溫控制器（2141)、水浴加熱 槽（2142) 〇 二氣化碳濃度控制模組（7)。 冷藏箱（32)〔培養基運送模組（3)〕。 氣壓控制箱（8)及廢液儲存桶（9)。 13 200825168 請參閱第三圖，該培養箱（2)另内設有滑軌（25)， 而微重力控制模組（21)包括有微重力控制單元（211)、 培養基換取機構（212)、管路清洗模組（213)〔圖中未 示〕。 又，微重力控制單元（211)固定於一基板（23)上， 基板（23)底部設有滑槽（24)，將基板（23)之滑槽（24) 組合於培養箱（2)之滑執（25)，使兩者可相對滑移。 而，培養基換取機構（212)係設有出水口（2124)、 • 進水口（2125)、擋板（2126)、第二氣壓缸（2127)， 該檔板（2126)係阻擋於出水口（2124)、進水口（2125) 之前端。 請參閱第四圖，係為加熱單元（214)之剖視圖，加 _ 熱單元（214)係設有水浴加熱槽（2142)及恆溫控制器 (2141);主要係將水浴加熱槽（2142)隔水加熱至所需 溫度，並將欲加熱之運送培養基之第三管路（31)繞行裝 _ 設於水浴加熱槽（2142)内，培養基通過此第三管路（31) 時將會被水浴加熱槽（2142)的水所加熱，並在水浴加熱 槽（2142)中加上攪拌葉片（2143)，使之產生渦流以快 速將水浴加熱槽（2142)内水的溫度加熱至所設定之溫度 值，培養基在水浴加熱槽（2142)中行走越長的距離越能 達到操作者所需求之溫度，而其所需之長度則端賴操作者 所需'之溫度而設計。 請參閱第五圖，恆溫模組（22)，包括有加熱控制器 200825168 (221)及,皿度感測益（ 222)，加熱控制器（Μ〗）係組 〇又於上述培養箱（2)之外園，而溫度感測$ (222)設於 培養箱（2)内部。 μ請再參閱第五圖，藉由溫度感測器（222)偵測培養 箱（2)内σρ之溫度，而將其溫度訊號傳回電控模組（4) 〔圖中未示〕，而可控制培養箱（2)内部之溫度變化。 而良胍模組（22)之作用係基於細胞體外培養需要創造出 有如人體-樣的環境’健康正常人體溫為们上下，故 在心養、.、田胞日守也需要有相同溫度環境，本發明係利用加熱 控制器（221)關於培養箱⑺之外目，祕傳方式將 溫度傳遞至培養箱（2)内部，而培養箱（2)内部裝設溫 -度感測器（222)以回授溫度訊號，藉由電控模組⑷進 -行監控。本發明所開發之怪溫模組（22)也可做適應性溫 度變化，可自行設定溫度範圍為室溫至4〇沱之間。 請參閱第六圖，其亦為微重力控制模組（21)設入培 籲養Is (2)後之第一動作」，微重力控制單元（m)係 藉由第一氣餘（2117)推出培養箱⑴外，直所在之、 位置為無菌操作室（5)〔圖中未示〕，操作者由無菌操 作室⑴之正Μ空間中裝入培養盤⑽2)，待培養盤 ,(2112)組裝完成，再由第—氣壓紅（2117)拉回培養箱 (2)内〔如第五圖所示〕’進入後由紅外蚊位器（21 校正培養盤㈤2)之第一快速接頭⑽3)、第二快速 接頭（2114)與培養基換取機構（212)之出水口（2124)、 200825168 進水口（2125)對準。 「第二動作」如第七、八圖所示，請先參閱第七圖， 本圖為培養基換取機構（212)〔圖中未示〕之擋板（2126) 擋住出水口（2124)、進水口（2125)之示意圖。 請參閱第八圖，令擋板（2126)退開，使擋板（2126) 不再阻擂出水口（2124)、進水口（2125)。 「第三動作」如第九圖所示，培養基換取機構（212) 由第二氣壓缸（2127)頂出，而使出水口（2124)插入第 ⑩ 一快速接頭（2113)，而進水口（2125)插入第二快速接 頭（2114)，當培養基注入完畢，或是置換完畢；「第四 動作」〔行程回到第五圖之狀態〕，第二氣壓缸（2127) - 再將培養基換取機構（212)拉回，不使用時擋板（2126) - 復位，再擋住出水口（2124)、進水口（2125)〔行程回 到第七圖之狀態〕，之後管路清洗模組（213)由第一幫 浦（2121)之第一管路（2122)經由三通閥（2123)，透 I 過第二管路（2131)引入去離子水，以去離子水清洗第一 管路（2122)、加熱單元（214)、培養基換取機構（212) 之出水口（2124)、進水口（2125)等内部管路，並利用 擋板（2126)防止清洗液直接喷出造成污染，在培養基換 取機構（212)下方管路清洗模組（213)另設有蓄液槽 (2132)將清洗後之廢去離子水收集並抽離培養箱（2) 至廢液儲存桶（9)。 另，當培養盤（2112)内之細胞需要更換培養基，培 16 200825168 養基換取麟⑽）需接合培養盤（2112) ^浦（2120即控難送冷藏箱（32)内之培養基， <基運运模組⑺之第三管路（31)經過 口 =:、第一管路(2122)進入培養基二 而當 第-幫浦(2121)控制管路1 =板:2126)復位時， 養基之所有内部管路(213) ^_培 生附著動作完成。 擁—直重複職個細胞增 腦係培養基運送示4®，係由控制電 經由動培養基換取機構（212)之第—幫浦（2d)， 冷藏二St且(3)之第三管路(31)將培養基由 送至加執β…工—通閥（2123)、第—管路（2122)運 再由^早儿⑵4)升溫’再送往培養基換取機構（212)， 盤基換取機構（212)之出水口⑽）透過培養 卷—於之第一快速接頭（2113)進入培養盤（2112)， 二速^當時間後培養基之養分耗盡，廢培養基則由第二 (212^ (2114)經由培養基換取機構（212)之進水口 養基，IT廢培養基送至廢液儲存摘(9)，此為培 基換第十1 ’係由係由控制電腦（43)啟動培養 )〔圖中未不〕之第二管路（2131)，再經由 17 200825168 二通閥（2123)、第一管路（2122)、加熱單元（21幻 —送往培養基換取機構（212)〔以清洗出水口（2124)、 進水口（2125)及内部管路〕，後將廢去離子水送至廢液 餘存桶（9)，此為去離子水之運送路徑之一。 Ά 請再參閱第十二圖，其係去離子水運之運送路徑之一 之示意圖，係由控制電腦（43)控制去離子水運送模組 之第二幫浦（61)，經由第四管路（62)將去無子水運、关 至水龍頭（ 52)，使用過後之廢去離子水流人水槽 > 再送至廢液儲存桶（9)。 ’ 本發月之第-幫浦、第二幫浦係為橋動式幫浦，如第 t三圖所示’勒式幫浦能有效的控制流量，^因是用押. 壓方式推動液體，因此管路較不易阻塞。當馬達轴心旋= .時，軸心周圍的滾輪壓住内有液體的管道，在兩個滚輪之 間的液體將隨著滾輪的移動而移動。由於細胞相當脆弱， 故於培養基更換時，蠕動式幫浦即發揮作用，馬達以不傷 #細胞之速度，藉由滚輪以漸進方式將經過加熱至適當溫度 之心養基推入培養盤内’耗盡養份之培養基則慢慢被推擠 而排出培養盤。 上述之—氣化石厌派度控制單元，其作用係在於控制培 •養^内之二氣化碳濃度，使其濃度趨近於人體血液内二氣 化反之辰度，使增生元成之細胞可適應人體以提高移植之 成功率。二氣化破濃度感測器所債測之二氧化碳濃度百分 比顯示於控制電腦上，以便於隨時監控，當濃度不足時電 200825168 磁閥將會開啟通入適量二氣化碳氣體調節，當濃度過濃時 將會通入乾淨空氣降低濃度，使細胞能隨時處於操作者設 定之濃度環境。 【圖式簡單說明】 第一圖係為本發明主要構件之方塊圖。 第二圖係為本發明主架體之立體外觀圖。 第三圖係為本發明加熱單元之剖視圖。 第四圖係為本發明微重力控制模組設入培養箱之示意 圖。 第五圖係為本發明恆溫模組之加熱控制器貼附於培養 箱之不意圖。 第六圖係為本發明微重力控制單元推出培養箱外之示 意圖。 第七圖係為本發明檔板擋住出水口、進水口之示意圖。 第八圖係為本發明擋板退開出水口、進水口之示意圖。 第九圖係為本發明培養基換取機構頂出，使出水口插入 第一快速接頭，進水口插入第二快速接頭之示意圖。 第十圖係為本發明培養基運送至培養基換取機構之示 意圖。 第十一圖係為本發明去離子水運送至培養基換取機構 之示意圖。 第十二圖係為本發明去離子水運送至水龍頭、水槽、廢 液儲存桶之示意圖。 19 200825168 第十三圖係為蠕動式幫浦之擠壓推動液體示意圖。 第十四圖係為習用傳統培養室之裝備示意圖。 【主要元件符號說明】The water bath is provided with a vane in the heating tank, and the third crucible in the water bath heating tank is placed in a bypass manner. The nucleus is heated to a temperature suitable for the length of the third conduit bypass described above to allow the cells to grow. Control unit, control power The above-mentioned electronic control module is equipped with a power supply. Brain 0 gasification carbon concentration control module The above-mentioned aseptic operation room indoor injury #古水槽, outdoor is equipped with a net, ventilation:. Sterilization light source, water dragonity sense = money material (four) garment two gasification; the second line of the above three-way ' second line 'other' three lines is open circuit I / third line name The first pump, the second pump system::: official road is the same * the above waste liquid storage tank ', two peristalic pump. It is used to recycle waste deionized water and waste grid 9 200825168. The invention has the following advantages: 1. The invention integrates various devices into one by control integration technology, controls the culture environment through sensing and quantitative components, and automates the complex exchange process of the culture medium and timely and quantitatively supplies the medicines, thereby making the cultivation process The factors that affect cell proliferation can be greatly reduced. When adverse conditions and cell proliferation conditions are reduced, the cells can be rapidly cultured in large quantities. 2. By the present invention, both planar and stereoscopic cell cultures are applicable, and their proliferative attachment state is more uniform. 3. The whole process of the invention is automated, which can reduce human error and pollution, and is beneficial to the cultivation of cells, and the quality and quantity of cells are greatly improved. 1 The invention integrates various devices into one, effectively saves indoor space, „ can effectively simplify the process and save cost. [Methods] The present invention is a microgravity aseptic environment cell automatic culture device, please refer to the first figure first. This drawing is the main component explanatory drawing of the present invention (not shown in the detailed part drawing), which is provided with the main frame body (1), and the main frame body (1) is provided with an incubator (2), and the medium is transported. Module (3), electronic control module (4), aseptic operation room (5), deionized water transport module (6), two gasification carbon concentration control module (7), pneumatic control box (8) and Waste liquid storage tank (9), wherein: " incubator (2), including microgravity control module (21) and thermostatic module (22); and microgravity control module (21) including microgravity Control 200825168 unit (211), medium exchange mechanism (212), pipeline cleaning module (213) 'In addition, the medium exchange mechanism (212) is provided with a first pump (2121) and a first pipeline (2122). And the first pipeline (2122) is engaged with the three-way valve (2123), and the pipeline cleaning module (213) is provided with the second pipeline (2131) The second pipeline (2131) of the liquid storage tank (2132) is also joined to the three-way valve (2123), and the thermostat module (22) is provided with a heating controller (221) and a temperature sensor (222). The heating controller (221) is disposed outside the incubator (2), and the azL sensing person (222) is disposed inside the incubator (2). In addition, the pipeline cleaning module (213) The second pipeline (2131) is connected to the ion water storage tank (n) of the main frame body (1), and the first pipeline (2122) of the medium exchange mechanism (212) is provided with a heating unit (214). The heating unit (214) is provided with a water bath heating-tank (2142) and a thermostat controller (2141), and the water bath heating tank (2142) is provided with blades (2143), and the water bath heating tank (2142) The first official road (2122) is placed in a bypass manner, and the length of the first conduit (2122) bypasses the medium to heat the medium to a temperature suitable for cell growth. In addition, the microgravity control unit (211) is provided with infrared rays. Positioner (2111), culture plate (2112), first quick connector (2113), first quick connector (2114), control box (2115), vent hole (2116), and first gas Cylinder (2117). The medium exchange mechanism (212) is further provided with a water outlet (2124), a water inlet (2125), a baffle (2126) and a second pneumatic cylinder (2127); a complex, microgravity control unit (211) Fixed on a substrate (23), the base 11 200825168 plate (23) is provided with a sliding slot (24) at the bottom, and the sliding groove (24) of the substrate (23) is combined with the sliding rail provided by the incubator (2) (25) ), so that the two can be relatively slippery. The medium transport module (3) is provided with a third pipeline (31), the third pipeline (31) is coupled to the three-way valve (2123), and is provided with a refrigerator (32) for preserving the medium and cultivating Reagent and cell growth factor; medium exchange mechanism (212) for transporting the medium through the third conduit (31) to the microgravity control module (21). The electric control module (4) is provided with a power supply (41), a control unit (42), a control computer (43), and the electronic control module (4) controls the computer (43) by human operation, and controls the computer ( 43) The control unit (42) monitors the constant temperature 'module (22), the two gasification carbon concentration control module (7), and the power supply and supply unit (41) supplies all components requiring power. The aseptic operation room (5) is provided with a UV germicidal light source (51), a faucet (52), a water tank (53), a filter screen (54), and an exhaust fan (55). The water φ tank (Μ) does not use the daily guard. Cover plate (not shown) is covered to enlarge the space available in the aseptic operation room (5); the function of the exhaust fan (55) is that if the door of the aseptic operation room (5) is opened, the exhaust fan (55) Immediately, the outside air is drawn into the aseptic operation room via the filter (54), and the external air drawn by the UV germicidal light source (51) is disinfected, so that the aseptic operation room (5) maintains the sterile space. And make the aseptic operation room (5曰) form a positive pressure space. Deionized water transport module (6), equipped with a second pump (61), 12 200825168 fourth pipeline (62) 'transported deionized water from the deionized water storage tank (11) to the aseptic operation room ( 5) The faucet (52). The second gasification carbon concentration control module (Ό, includes a two gasification carbon concentration sensor (71), a solenoid valve (72). The air pressure control box (8) is controlled by the microgravity control unit (211) The first pneumatic cylinder (2117) and the second pneumatic cylinder (2127) of the medium exchange mechanism (212). The waste liquid storage tank (9) is used for recovering waste deionized water and waste medium. • As shown in the second figure, It is a configuration diagram of the main frame body (1) and the above various components. It can be seen in the figure: Main frame body (1) - Incubator (2) Power supply module of electric control module (4) (41 ), control unit (42), control computer (43) Aseptic operation room (5) is equipped with UV germicidal light source (51), water tap (52), sink (53), and filter screen (54). ), exhaust fan (55) 〇 heating unit (214) [added to the third line (31) of the medium transport module (3)], equipped with a thermostat controller (2141), water bath heating tank (2142) 〇2 gasification carbon concentration control module (7). Refrigerator (32) [media transport module (3)] Pneumatic control box (8) and waste storage tank (9). 13 200825168 Referring to the third figure, the incubator (2) is further provided with a slide rail (25), and the microgravity control module (21) comprises a microgravity control unit (211), a medium exchange mechanism (212), and a pipeline cleaning. The module (213) (not shown). The microgravity control unit (211) is fixed on a substrate (23), and the bottom of the substrate (23) is provided with a sliding slot (24) for sliding the substrate (23). The trough (24) is combined with the slipper (25) of the incubator (2) so that the two can be relatively slipped. However, the medium exchange mechanism (212) is provided with a water outlet (2124), a water inlet (2125), The baffle (2126) and the second pneumatic cylinder (2127) are blocked at the front end of the water outlet (2124) and the water inlet (2125). Referring to the fourth figure, the heating unit (214) In the cross-sectional view, the heating unit (214) is provided with a water bath heating tank (2142) and a thermostat controller (2141); the main purpose is to heat the water bath heating tank (2142) to a desired temperature, and to heat the transport. The third line (31) of the medium is arranged in the water bath heating tank (2142), and the medium will be heated by the water bath when passing through the third line (31). The water of the tank (2142) is heated, and a stirring blade (2143) is added to the water bath heating tank (2142) to generate a vortex to rapidly heat the temperature of the water in the water bath heating tank (2142) to the set temperature value. The longer the medium travels in the water bath heating tank (2142), the higher the temperature required by the operator, and the required length is designed according to the temperature required by the operator. Please refer to the fifth figure. The thermostat module (22) includes a heating controller 200825168 (221) and a sense sensor (222), a heating controller (Μ), and a temperature group outside the incubator (2), and the temperature Sensing $ (222) is located inside the incubator (2). μ Please refer to the fifth figure. The temperature sensor (222) detects the temperature of σρ in the incubator (2) and transmits its temperature signal back to the electronic control module (4) (not shown). The temperature change inside the incubator (2) can be controlled. The role of Liangzhu module (22) is based on the need to create a human-like environment based on the in vitro culture of cells. The healthy and normal body temperature is up and down. Therefore, it is necessary to have the same temperature environment in the heart, and the field cell. The invention utilizes a heating controller (221) to transfer the temperature to the inside of the incubator (2) in a secret manner with respect to the incubator (7), and a temperature-degree sensor (222) is installed inside the incubator (2) to The temperature signal is feedbacked and monitored by the electronic control module (4). The strange temperature module (22) developed by the invention can also be adapted to the temperature change, and can be set to a temperature ranging from room temperature to 4 自行. Please refer to the sixth figure, which is also the first action after the microgravity control module (21) is set up in the training (I). The microgravity control unit (m) is based on the first gas (2117). The incubator (1) is placed outside, and the position is the aseptic operation room (5) (not shown). The operator loads the culture tray (10) 2) from the orthodontic space of the aseptic operation room (1), and the tray is to be cultured (2112). The assembly is completed, and then the first pressure connector (10) 3 is pulled back into the incubator (2) by the first pressure red (2117) [as shown in the fifth figure] after entering the infrared mosquito device (21 calibration plate (5) 2). The second quick connector (2114) is aligned with the water outlet (2124) of the medium exchange mechanism (212) and the water inlet (2125) of the 200825168. "Second action", as shown in the seventh and eighth figures, please refer to the seventh figure. This picture shows the baffle (2126) of the medium exchange mechanism (212) (not shown) blocking the water outlet (2124) and entering Schematic diagram of the nozzle (2125). Refer to Figure 8 to disengage the baffle (2126) so that the baffle (2126) no longer blocks the water outlet (2124) and the water inlet (2125). "Third action" As shown in the ninth figure, the medium exchange mechanism (212) is ejected by the second pneumatic cylinder (2127), and the water outlet (2124) is inserted into the 10th quick joint (2113), and the water inlet ( 2125) Insert the second quick connector (2114), when the medium is injected, or the replacement is completed; "fourth action" (the stroke returns to the state of the fifth figure), the second pneumatic cylinder (2127) - the medium is exchanged for the mechanism (212) Pull back, when not in use, the baffle (2126) - reset, then block the water outlet (2124), the water inlet (2125) [stroke back to the state of the seventh figure], and then the pipeline cleaning module (213) The first line (2122) of the first pump (2121) is introduced into the deionized water through the second line (2131) via the three-way valve (2123), and the first line is cleaned with deionized water (2122) ), the heating unit (214), the water outlet (2124) of the medium exchange mechanism (212), the water inlet (2125) and other internal pipelines, and the baffle (2126) is used to prevent the direct injection of the cleaning liquid to cause pollution, and the medium is exchanged. The pipeline cleaning module (213) under the mechanism (212) is further provided with a liquid storage tank (2132) to be cleaned. Deionized water was collected and detached incubator (2) to a waste storage barrels (9). In addition, when the cells in the culture plate (2112) need to be replaced with medium, the culture medium (2112) and the medium (2120) are controlled to be transported in the refrigerator (32), <2120; The third line (31) of the base transport module (7) passes through the port =:, the first line (2122) enters the medium 2, and when the first pump (2121) control line 1 = board: 2126) is reset, All internal piping of the nutrient base (213) ^_Peisheng attachment action is completed. Convenient-repetitive cell-enhanced medium delivery medium 4® is controlled by the control medium via the dynamic medium exchange mechanism (212) - pump (2d), refrigerated two St and (3) third line ( 31) The culture medium is sent to the addition of the β-work valve (2123), the first line (2122), and then heated by the early (2) 4) and sent to the medium exchange mechanism (212), the disk exchange mechanism (212) The water outlet (10)) passes through the culture volume - the first quick connector (2113) enters the culture plate (2112), the second speed ^ after the time, the culture medium is depleted, and the waste medium is second (212^ ( 2114) via the medium exchange mechanism (212) inlet nutrient base, IT waste medium is sent to the waste liquid storage pick (9), which is the tenth 1 'system of the base is controlled by the control computer (43)) The second line (2131) of the figure is not the same, and then through the 17 200825168 two-way valve (2123), the first line (2122), the heating unit (21 phantom - sent to the medium exchange mechanism (212) [to clean The water outlet (2124), the water inlet (2125) and the internal pipeline], and then the waste ionized water is sent to the waste liquid storage tank (9), which is deionized. One of the water transport routes. Ά Please refer to the twelfth figure, which is a schematic diagram of one of the transport paths of deionized water transport, which is controlled by the control computer (43) to control the second pump of the deionized water transport module (61). ), through the fourth pipeline (62), the water is transported to the faucet (52), and the used deionized water flow tank is used, and then sent to the waste storage tank (9). -The pump and the second pump are bridge-type pumps. As shown in the figure t, the 'Leh-type pump can effectively control the flow, ^ is because the pressure is used to push the liquid, so the pipeline is not easy. Blocking. When the motor shaft is rotating, the roller around the shaft presses the pipe with the liquid inside, and the liquid between the two rollers will move with the movement of the roller. Since the cells are quite fragile, the medium is replaced. At the same time, the peristaltic pump works, and the motor pushes the heart-feeding base heated to the appropriate temperature into the culture tray by the roller in a progressive manner without damaging the speed of the cell. Being pushed out and discharging the culture plate. The above-mentioned gas fossil annoyance control The function of the unit is to control the concentration of the two gasified carbon in the culture and the nutrient, so that the concentration is close to the second gasification in the human blood, and the cell of the proliferator can adapt to the human body to improve the success of the transplant. The percentage of carbon dioxide concentration measured by the two gasification and concentration sensor is displayed on the control computer so as to be monitored at any time. When the concentration is insufficient, the 200825168 magnetic valve will be opened and the appropriate amount of two gasification carbon gas will be adjusted. When the concentration is too rich, clean air will be introduced to reduce the concentration, so that the cells can be in the concentration environment set by the operator at any time. [Simplified illustration] The first figure is a block diagram of the main components of the invention. The second figure is a three-dimensional appearance of the main frame body of the present invention. The third figure is a cross-sectional view of the heating unit of the present invention. The fourth figure is a schematic diagram of the microgravity control module of the present invention being placed in an incubator. The fifth figure is not intended to be attached to the incubator of the heating controller of the thermostat module of the present invention. The sixth figure is an indication of the introduction of the microgravity control unit of the present invention outside the incubator. The seventh figure is a schematic view of the baffle of the invention blocking the water outlet and the water inlet. The eighth figure is a schematic view of the baffle retreating the water outlet and the water inlet of the present invention. The ninth figure is a schematic diagram of the medium exchange mechanism of the present invention ejecting, the water outlet is inserted into the first quick joint, and the water inlet is inserted into the second quick joint. The tenth figure is an illustration of the transport of the medium of the present invention to the medium exchange mechanism. The eleventh figure is a schematic view of the transport of deionized water to the medium exchange mechanism of the present invention. Figure 12 is a schematic view showing the delivery of deionized water to a faucet, a water tank, and a waste liquid storage tank of the present invention. 19 200825168 The thirteenth picture is a schematic diagram of the push-pull liquid of the peristaltic pump. The fourteenth figure is a schematic diagram of the equipment used in the conventional culture room. [Main component symbol description]
(1) 主架體 (2) 培養箱 (211) 微重力控制單元 (2112)培養盤 (2114)第二快速接頭 (2116)通氣孔 (212) 培養基換取機構 (2122)第一管路 (2124)出水口 (2126)檔板 (213) 管路清洗模組 (2132)蓄液槽 (214) 加熱單元 (2142)水浴加熱槽 (22) 恆溫模組 (222)溫度感測器 (23) 基板 (25)滑執 (3) 培養基運送模組 (32)冷藏箱 (11) 去離子水儲存桶 (21) 微重力控制模組 (2111) 紅外線定位器 (2113) 第一快速接頭 (2115) 控制盒 (2117) 第一氣壓缸 (2121) 第一幫浦 (2123) 三通閥 (2125) 進水口 (2127) 第二氣壓缸 (2131) 第二管路 (2141) 恆溫控制器 (2143) 葉片 (221) 加熱控制器 (24) 滑槽 (31) 弟二管路 20 200825168(1) Main frame body (2) Incubator (211) Microgravity control unit (2112) Culture plate (2114) Second quick connector (2116) Vent hole (212) Medium exchange mechanism (2122) First pipe (2124) ) Water outlet (2126) baffle (213) Pipe cleaning module (2132) Storage tank (214) Heating unit (2142) Water bath heating tank (22) Thermostatic module (222) Temperature sensor (23) Substrate (25) Slipper (3) Medium transport module (32) Freezer (11) Deionized water storage tank (21) Microgravity control module (2111) Infrared locator (2113) First quick connector (2115) control Box (2117) First pneumatic cylinder (2121) First pump (2123) Three-way valve (2125) Inlet (2127) Second cylinder (2131) Second line (2141) Thermostat (2143) Blade (221) Heating controller (24) chute (31) brother two pipeline 20 200825168
(4) 電控模組 (41) 電源供應器 (42) 控制單元 (43) 控制電腦 (5) 無菌操作室 (51) UV殺菌光源 (52) 水龍頭 (53) 水槽 (54) 濾網 (55) 抽風機 (6) 去離子水運送模組（61) 第二幫浦 (62) 第四管路 (7) 二氣化碳濃度控制模組 (71) 二氣化碳濃度感測器 (72) 電磁閥 (8) 氣壓控制箱 (9) 廢液儲存桶 (AD 水池 (A2) 晾乾架 (A3) 消毒器 (A4) 無菌操作室 (A5) 準備室 (A6) 電冰箱 (A7) 離心機 (A8) 恆溫培養箱 (A9) 顯微鏡天平台 (A10) 實驗台 (All) 污物桶 (A12) 藥品櫃 (A13) 器械櫃 21(4) Electronic control module (41) Power supply (42) Control unit (43) Control computer (5) Aseptic operation room (51) UV germicidal light source (52) Faucet (53) Sink (54) Filter (55 Exhaust fan (6) Deionized water transport module (61) Second pump (62) Fourth pipeline (7) Two gasification carbon concentration control module (71) Two gasification carbon concentration sensor (72 Solenoid valve (8) Air pressure control box (9) Waste liquid storage tank (AD pool (A2) Drying rack (A3) Sterilizer (A4) Aseptic operation room (A5) Preparation room (A6) Refrigerator (A7) Centrifugation Machine (A8) Constant Temperature Incubator (A9) Microscope Sky Platform (A10) Test Bench (All) Waste Bucket (A12) Medicine Cabinet (A13) Instrument Cabinet 21