1337621 .•九、發明說明： 【發明所屬之技術領域】 - 本發明是有關於一種生物反應系統’特別是有關於一 . 種細胞分離裝置。 【先前技術】 一般來說’目前一些細胞培養系統通常必須應用特定 的固液分離裝置來將所培養出的細胞分離出來。也就是 說，固液分離裝置可將細胞從代謝液中分離出來，而所分 • 離出之細胞可被輸出，應用或被回收去進行進一步的增生培1337621 . IX. DESCRIPTION OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to a biological reaction system, and more particularly to a cell separation device. [Prior Art] Generally, some cell culture systems currently have to use a specific solid-liquid separation device to separate the cultured cells. That is to say, the solid-liquid separation device can separate cells from the metabolic fluid, and the separated cells can be exported, applied or recycled for further proliferation.
就細胞分離技術而言，一種習知之固液分離裝置是利 用過濾膜來將細胞從代謝液中分離出來。換言之，當細胞 及代謝液流經過濾膜時，細胞會被過濾膜所攔戴，:代謝 液會通過過濾膜。然而，由於代謝液中會含有微小的細胞 代謝產物，故在過濾膜經長時間使用後，細胞代謝產物常 會堆積於過濾膜上，因而會造成過濾膜阻塞。 吳國專利第5,817,505號揭露了一種用來分離細胞之 固液分離裝置’其具有傾斜式多層平板之構造，以克服過 遽膜阻塞_題。細胞及代職可藉由财幫浦之抽打而 由下往上地流過傾斜式多層平板。在此，由於細胞之重量 (或比重）較大以及代謝液之重f (或比重）較小，故細胞會受 重力影響而沉降至固液分離裝置（或傾斜式多層平板）之底 部’而代職齡被騎⑽㈣分離裝置（絲斜式多層 千板）。S而，以此種習知之固液分離裝置來分離細胞會具 1337621 了：點。百先’在細胞從傾斜式多層平板上沉降之過 二：.丨於：胞與傾斜式多層平板間之接觸面積小，故細 剪應力會較高。因此，細胞結構常會受此種高 =用：衷。其次，由於固液分離裝置(或傾斜式 心f質所製成’故在重複使用上需以高 ，皿μ式请毒’因而會導致高作業成本。再者，細胞 液㈣裝置内之冗長沉降過程中常會因缺氧而死亡， 因而，造成代謝物污染，進而不利於細胞輪出應 【發明内容】 ^發明係包括—細胞培養裝置，容納有複數個細胞及 一代谢液；一細胞分離裝置，具有-第-端部、一第二端 式圓弧管體’其中，該傾斜式圓弧管體係連接 部與該第二端部之間’以及該第二端部之水平 向度係大於該第一端部之水平高度；-第-輸送管，連通 於該細胞培養裝置與該細胞分離裝置之該第—端部之 一代謝液收集容器；_笛-私.、，μ 4 曰’ ^ν 第一輸迗s，連通於該代謝液收集 合态人5玄、，.田胞/刀離裝置之該第二端部之間，·一第一 設置於該第二輪送管中，其中，該細胞培養裝置中之該 細胞及該代謝液係藉由該第一幫浦之抽打而經由該[輸 送管輸送至該細胞分離裝置之中，該等細胞係經由該傾： ^圓弧管體而沉降至該第一端部，以及該代謝液係經由該 第一輸达官而輸送至該代謝液收集容器之中；—第三 管，連通於該細胞培養I置與該細胞分離裝置之該第= 部之間；以及-第二幫浦，設置於該第三輸送管中，其中， UJ7621 4於1亥第端部之該等細胞係藉由該第二幫浦之抽打而經 由該第三輸送管輪送至該細胞培養裝置之中。 同恰，根據本發明之生物反應系統，該細胞分離裝置 更具有至少一個圓弧層板，以及該等圓弧層板係設置於該 倾斜式圓弧官體之中，並且係連接於該第一端部與該第二 端部之間。 管二在本發明中’該等圓弧層板係平行於該傾斜式圓弧In the case of cell separation techniques, a conventional solid-liquid separation device utilizes a filtration membrane to separate cells from a metabolic fluid. In other words, when cells and metabolic fluids flow through the filter membrane, the cells are blocked by the filter membrane: the metabolic fluid passes through the membrane. However, since the metabolic fluid contains minute cellular metabolites, after the filter membrane is used for a long period of time, cellular metabolites often accumulate on the filter membrane, which may cause the membrane to clog. U.S. Patent No. 5,817,505 discloses a solid-liquid separation device for separating cells, which has a configuration of a tilted multi-layered plate to overcome over-membrane obstruction. The cells and the substitutes can flow through the inclined multi-layer slab from bottom to top by the slap of the gang. Here, since the weight (or specific gravity) of the cells is large and the weight f (or specific gravity) of the metabolic fluid is small, the cells are settled by gravity to settle to the bottom of the solid-liquid separation device (or the inclined multi-layer plate). The off-age is riding (10) (four) separation device (silk-slope multi-layer board). In contrast, the conventional solid-liquid separation device for separating cells will have 1337621 points: The first shearing stress on the cells is from the inclined multi-layer plate. The contact area between the cells and the inclined multi-layer plate is small, so the fine shear stress will be high. Therefore, the cell structure is often subject to this high = use: sincerity. Secondly, due to the solid-liquid separation device (or the inclined core is made of 'therefore, it needs to be high in repeated use, and the dish is poisoned by the dish type'), which leads to high operating cost. Furthermore, the cytosol (4) is long in the device. In the process of sedimentation, it often dies due to lack of oxygen, thus causing metabolite contamination, which is not conducive to cell rotation. [Invention] The invention includes a cell culture device containing a plurality of cells and a metabolic fluid; The device has a first end portion and a second end arc tube body, wherein a horizontal dimension between the inclined arc tube system connection portion and the second end portion and the second end portion a level larger than the first end portion; a first-transport tube connected to the cell culture device and the first end of the cell separation device; the metabolic fluid collection container; _ flute-private.,, μ 4 曰' ^ ν first 迗 s, connected to the metabolic fluid to collect the conjugated person 5 玄 , , between the second end of the field cell / knife off device, · a first set in the second wheel Wherein the cell in the cell culture device and the metabolic fluid system are borrowed The first pump is pumped through the [transport tube to the cell separation device, and the cell lines are sedimented to the first end via the tilting: ^ arc tube body, and the metabolic liquid system Transmitting to the metabolic fluid collection container via the first delivery officer; - a third tube connected between the cell culture I and the third portion of the cell separation device; and - a second pump, setting In the third transport tube, wherein the cell lines of the UJ7621 at the first end of the 1H are transferred to the cell culture device via the third transfer tube by the second pump. According to the biological reaction system of the present invention, the cell separation device further has at least one circular arc layer, and the circular arc plate is disposed in the inclined arc body and is connected to the first Between the end portion and the second end portion. In the present invention, the arc-shaped layer plate is parallel to the inclined arc
μ又在本發明中，該等圓弧層板之曲率係與該傾斜式圓 弧管體之曲率相同。 又在本發明中，該等圓弧層板間之距離係介於1公分 與4公分之間。 又在本發明中，該傾斜式圓弧管體及該等圓弧層板係 由聚丙烯所製成。 又在本發明中，該傾斜式圓弧管體係由聚丙烯所製成。 又在本發明中，該傾斜式圓弧管體與一水平面間之傾 斜角度係介於20度與60度之間。 ^又在本發明中，更包括一震盪器，係設置於該細胞分 離裝置之上，用以震盪沉降該等細胞。 又在本發明中，更包括一氧氣供應裝置，係連通於該 細胞培養裝置及該細胞分離裝置。 又在本發明中，該氧氣供應裝置係連通於該細胞分離 裝置之該第二端部。 又在本發明中，更包括一溫度調節裝置，係設置於該 1337621 .第一輸送管之上，用以調節通過該第一輸送管之該等細胞 及該代謝液之溫度。. - 又在本發明中更包括一培養液補充裝置，係連通於 該細胞坨養裴置，用以將一培養液補充至該細胞培養裝置 之中。 為使本發明之上述目的、特徵和優點能更明顯易懂， 下文特舉較佳實施例並配合所附圖式做詳細說明。 【實施方式】 兹配合圖式說明本發明之較佳實施例。 請參閱第1圖，本實施例之生物反應系統1〇〇主要包 括有一細胞培養裝置〗10、一細胞分離裝置120、一第一輪 送管、一代謝液收集容器140、一第二輸送管132、一 第一幫浦151、一第三輸送管133、一第二幫浦152、一震 盪态160、一氧氣供應裝置17〇、一溫度調節裝置]8〇及_ 培養液補充裝置190。 細胞培養裝置11〇内容納有複數個細胞C及一代謝液 Μ。在此’代谢液μ大致上是由細胞代謝產物及培養液所 組成。此外’細胞培養裝置η〇具有一攪拌器〗η。藉由 授拌β 111之攪拌，細胞C即可均勻地分佈於細胞培養襞 置110之中，進而有利於其增生培養。 請參閱第1圖、第2圖及第3Α圖，細胞分離裝置120 具有一第一端部121、一第二端部122、一傾斜式圓弧管體 123及至少一個圓弧層板124。在本實施例中，該傾斜是圓 弧官體123具有複數個圓弧層板124。 1337621 第二端部122是相對於第一端部121。特別的是，第 二端部]22之水平高度是大於第一端部121之水平高度。 傾斜式圓弧管體123是連接於第一端部12〗與第二端 部122之間。在本實施例之中，傾斜式圓弧管體123與一 水平面間之傾斜角度可以是介於20度與60度之間。 複數個圓弧層板124是設置於傾斜式圓弧管體123之 中，並且複數個圓弧層板124亦是連接於第一端部121與 第二端部122之間。在本實施例之中，複數個圓弧層板]24 皆是平行於傾斜式圓弧管體123,或者複數個圓弧層板124 之曲率是與傾斜式圓弧管體123之曲率相同。此外，複數 個圓弧層板124彼此間之距離可以是介於1公分與4公分 之間，以及傾斜式圓弧管體123及複數個圓弧層板124可 以是由聚丙烯等生物相容性材料所製成。另外，值得注意 的是，本實施例之細胞分離裝置120並不侷限於具有半管 形之傾斜式圓弧管體123。舉例來說，如第3B圖所示，細 胞分離裝置120亦可以具有1/4管形之傾斜式圓弧管體 123,。 請參閱第1圖，第一輸送管131是連通於細胞培養裝 置110與細胞分離裝置120之第一端部121之間。 第二輸送管132是連通於代謝液收集容器140與細胞 分離裝置120之第二端部122之間。 第一幫浦]51是設置於第二輸送管132中。 第三輸送管133是連通於細胞培養裝置110與細胞分 離裝置120之第一端部121之間。 1337621 第二幫浦152是設置於第三輸送管丨33中。 震蘯裔〗60是設置於細胞分離裝置12〇之上。 、氧氣供應裝置.170係連通於細胞培養裝置11〇及細胞 '二離裝置I20。在本實施例之中，氧氣供應裝置】70乃是 藉由一第四輸送管134來連通於細胞分離裝置〗2〇之第二 端部122。 溫度調節裝置]80是設置於第一輪送管131之上。 坨養液補充裝置19〇是連通於細胞培養裝置。更 #詳細的來說，培養液補充裝置190是藉由一第五輸送管135 來連通於細胞培養裝置11〇，以及一第三幫浦153是設置 於第五輸送管135中。因此，藉由第三幫浦153之抽打， 培養液補充裝置190中之培養液即可經由第五輸送管135 補充至細胞培養裝置110之中。 如上所述，細胞培養裝置〗中之複數個細胞c及代 謝液Μ乃是II由第—幫浦151之抽打而經由第一輸送管 籲13^輸送至細胞分離裝置120之中。在此，由於細胞〇之 重里（或比重）較大以及代謝液Μ之重量（或比重）較小，故 細胞C會文重力影響而經由傾斜式圓弧管體123及複數個 圓弧層板124沉降至細胞分離裳置12〇之第一端部⑵， 、代4液Μ則會經由第二輸送管}來被抽打輸送至代謝 S收集容器⑽之中。特別的是，在細胞C從傾斜式圓弧 g版123及複數個圓弧層板124上沉降之過程中，由於細 胞c與傾斜式圓弧管體123或圓弧層板124間之接觸面積 已大幅增加（面接觸之形式），故細胞C所受到之剪應力會In yet another aspect of the invention, the curvature of the arcuate laminate is the same as the curvature of the inclined arcuate tubular body. Also in the present invention, the distance between the arc-shaped plates is between 1 cm and 4 cm. Further, in the present invention, the inclined arc tube body and the arc-shaped layer sheets are made of polypropylene. Also in the present invention, the inclined circular arc tube system is made of polypropylene. Further, in the present invention, the inclined angle between the inclined arc tube body and a horizontal plane is between 20 degrees and 60 degrees. Further, in the present invention, an oscillator is further disposed on the cell separation device for oscillating and sedimenting the cells. Further, in the present invention, an oxygen supply device is further included, which is connected to the cell culture device and the cell separation device. In still another aspect of the invention, the oxygen supply device is in communication with the second end of the cell separation device. Further, in the present invention, a temperature adjusting device is further disposed on the first transfer tube for adjusting the temperature of the cells and the metabolic liquid passing through the first delivery tube. Further, in the present invention, a culture solution replenishing device is further provided which is connected to the cell nurturing device for replenishing a culture solution into the cell culture device. The above described objects, features and advantages of the present invention will become more apparent from the description of the appended claims. [Embodiment] A preferred embodiment of the present invention will be described with reference to the drawings. Referring to FIG. 1 , the biological reaction system 1 of the present embodiment mainly includes a cell culture device 10 , a cell separation device 120 , a first wheel delivery tube, a metabolic fluid collection container 140 , and a second delivery tube. 132. A first pump 151, a third duct 133, a second pump 152, a oscillating state 160, an oxygen supply device 17A, a temperature adjusting device, and a cultivating fluid replenishing device 190. The cell culture device 11 contains a plurality of cells C and a metabolic fluid. Here, the 'metabolism fluid μ' is roughly composed of a cell metabolite and a culture solution. Further, the cell culture apparatus η has an agitator η. By stirring the β 111, the cells C can be uniformly distributed in the cell culture device 110, thereby facilitating the proliferation culture. Referring to Figures 1, 2 and 3, the cell separation device 120 has a first end portion 121, a second end portion 122, an inclined circular arc tube body 123 and at least one circular arc layer plate 124. In the present embodiment, the inclination is that the circular arc body 123 has a plurality of circular arc plates 124. 1337621 The second end 122 is opposite the first end 121. In particular, the level of the second end portion 22 is greater than the level of the first end portion 121. The inclined arc tube body 123 is connected between the first end portion 12 and the second end portion 122. In the present embodiment, the inclination angle between the inclined arc tube body 123 and a horizontal plane may be between 20 degrees and 60 degrees. A plurality of circular arc plates 124 are disposed in the inclined circular arc tube 123, and a plurality of circular arc plates 124 are also connected between the first end portion 121 and the second end portion 122. In the present embodiment, the plurality of circular arc plates 24 are parallel to the inclined circular arc tube 123, or the curvature of the plurality of circular arc plates 124 is the same as the curvature of the inclined circular arc tube 123. In addition, the plurality of circular arc plates 124 may be between 1 cm and 4 cm apart, and the inclined arc tube body 123 and the plurality of arc layer plates 124 may be biocompatible with polypropylene or the like. Made of sexual materials. Further, it is to be noted that the cell separation device 120 of the present embodiment is not limited to the inclined circular arc tube body 123 having a semi-tubular shape. For example, as shown in Fig. 3B, the cell separation device 120 may also have a 1/4 tube-shaped inclined arc tube body 123. Referring to Fig. 1, the first delivery tube 131 is connected between the cell culture device 110 and the first end portion 121 of the cell separation device 120. The second delivery tube 132 is in communication with the metabolic fluid collection container 140 and the second end 122 of the cell separation device 120. The first pump 51 is disposed in the second duct 132. The third delivery tube 133 is in communication with the cell culture device 110 and the first end portion 121 of the cell separation device 120. 1337621 The second pump 152 is disposed in the third duct 丨33. The shocking person 60 is placed on the cell separation device 12〇. The oxygen supply device .170 is connected to the cell culture device 11 and the cell 'dimerization device I20. In the present embodiment, the oxygen supply means 70 is connected to the second end portion 122 of the cell separation device by a fourth delivery tube 134. The temperature adjustment device 80 is disposed above the first transfer tube 131. The nutrient replenishing device 19 is connected to the cell culture device. More specifically, the culture solution replenishing device 190 is connected to the cell culture device 11 by a fifth delivery tube 135, and a third pump 153 is disposed in the fifth delivery tube 135. Therefore, the culture solution in the culture solution replenishing device 190 can be replenished into the cell culture device 110 via the fifth delivery tube 135 by the pumping of the third pump 153. As described above, the plurality of cells c and the sputum sputum in the cell culture apparatus are transported by the first pump 151 to the cell separation device 120 via the first delivery tube. Here, since the cell weight (or specific gravity) is large and the weight (or specific gravity) of the metabolic liquid is small, the cell C affects the gravity and passes through the inclined arc tube 123 and the plurality of circular arc plates. 124 is settled until the first end (2) of the cell separation is placed, and the liquid 4 is transported to the metabolic S collection container (10) via the second delivery tube. In particular, in the process of sedimentation of the cell C from the inclined arc g plate 123 and the plurality of arc plates 123, the contact area between the cell c and the inclined arc tube 123 or the circular arc plate 124 Has been greatly increased (in the form of face contact), so the shear stress of cell C will be
12 1337621 大幅減小，如此一來，細胞c之結構會較不易在沉降過程 中遭受破壞。此外，藉由震Μ II 16G之震盧作用，位於傾 斜式圓弧管體123’及複數個圓弧層板124上之細胞c即可 更快速地沉降至細胞分離裝置120之第一端部121。再者， 由於氧氣供應裝置17G可將氧氣供應至細胞分離裝置 之中’故細胞C便不會在沉降過程中因缺氧而死亡，如此 -來’位於細胞分離裝置12G中之細胞c的存活率即可提 ，。另外，溫度調節裝置18〇可適當地調節通過第一輸送 =131之細胞C及代謝液M之溫度，使細胞c不會在輸 送及沉降分離過程中改變溫度而造成死亡。 在另一方面，沉降至細胞分離裝置ί2〇之第一端部 之細胞C可藉由第二幫浦152之抽打而經由第三輸送管 133輸送（或回收）至細胞培養裝置】1〇之中，以進 + 地增生培養。 7 綜上所述，本發明所揭露之生物反應系統可具有以下 的優點： 〃12 1337621 is greatly reduced, so that the structure of cell c will be less susceptible to damage during sedimentation. In addition, the cell c located on the inclined arc tube body 123' and the plurality of arc layer plates 124 can be more quickly settled to the first end of the cell separation device 120 by the shocking effect of the shock absorber II 16G. 121. Furthermore, since the oxygen supply device 17G can supply oxygen into the cell separation device, the cell C does not die due to hypoxia during sedimentation, so that the survival of the cell c located in the cell separation device 12G The rate can be mentioned. Further, the temperature adjusting means 18 can appropriately adjust the temperature of the cells C and the metabolic liquid M passing through the first transport = 131 so that the cells c do not change in temperature during the transport and sedimentation separation to cause death. On the other hand, the cells C which have settled to the first end of the cell separation device can be transported (or recovered) to the cell culture device via the third transfer tube 133 by the pumping of the second pump 152. In the middle, it is cultivated by hyperplasia. In summary, the bioreactor system disclosed in the present invention can have the following advantages:
⑴可有效降低細胞在沉降分離過程中所受到的剪應力 作用’因而可提升細胞回收之品質。 本 (2)傾斜式圓弧管體及圓弧層板是由聚㈣等生物相容 ^材料所製m整個細胞分離裝置可在使用後直接抛 >、口而可免除％、複的 > 肖毒作業程序’進而可節省作業成 因而 (3)細胞不會在冗長的沉降過程中出現缺氧狀況 可有效提升細胞之存活率。(1) It can effectively reduce the shear stress that cells undergo during sedimentation and separation, thus improving the quality of cell recovery. The (2) inclined arc tube body and the arc layer plate are made of a biocompatible material such as poly (IV). The whole cell separation device can be directly thrown after use, and the mouth can be exempted from % and complex. The Xiaodu operation program can further save the operation. Therefore, (3) cells will not be hypoxic during the long sedimentation process, which can effectively improve the survival rate of the cells.
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雖然本發明已以較佳實施例揭露於上，然其並非用以 限定本發明，任何熟習此項技藝者，在不脫離本發明之精 神和範圍内，當可作些許之更動與潤飾，因此本發明之保 護範圍當視後附之申請專利範圍所界定者為準。 14 1337621 【圖式簡單說明】 .第1圖係、顯示本發明之生物反應系統之平面構造示意 圖， 第2圖係顯示本發明之生物反應系統之細胞分離裝置 之立體分解示意圖； 第3A圖係顯示本發明之生物反應系統之細胞分離裝 置之一種傾斜式圓弧管體之立體示意圖；以及 第3B圖係顯示本發明之生物反應系統之細胞分離裝 置之另一種傾斜式圓弧管體之立體示意圖。 【主要元件符號說明】 100〜生物反應系統 110〜細胞培養裝置 in〜攪拌器 120〜細胞分離裝置 121〜第一端部 〆 122〜第二端部 123、123’〜傾斜式弧管體 124〜圓弧層板 1W籮一輸送管w 132〜第二輸送管 133〜第三輸送管 134〜第四輸送管 135〜第五輸送管 15 1337621 140〜代謝液收集容器 151〜第一幫浦 152〜第二幫浦， 153〜第三幫浦 160〜震盪器 170〜氧氣供應裝置 180〜溫度調節裝置 190〜培養液補充裝置 c~細胞 Μ〜代謝液Although the present invention has been disclosed in its preferred embodiments, it is not intended to limit the present invention, and it is possible to make some modifications and refinements without departing from the spirit and scope of the present invention. The scope of the invention is defined by the scope of the appended claims. 14 1337621 BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a schematic view showing the planar structure of the biological reaction system of the present invention, and Fig. 2 is a perspective exploded view showing the cell separation device of the biological reaction system of the present invention; A perspective view showing a tilted circular arc tube of the cell separation device of the biological reaction system of the present invention; and FIG. 3B is a perspective view showing another inclined arc tube of the cell separation device of the biological reaction system of the present invention. schematic diagram. [Description of main component symbols] 100 to biological reaction system 110 to cell culture device in to agitator 120 to cell separation device 121 to first end portion 122 to second end portion 123, 123' to inclined arc tube body 124~ Arc layer plate 1W 输送 one conveying pipe w 132 〜 second conveying pipe 133 〜 third conveying pipe 134 〜 fourth conveying pipe 135 〜 fifth conveying pipe 15 1337621 140 ~ metabolic liquid collecting container 151 ~ first pump 152 ~ Second pump, 153 ~ third pump 160 ~ oscillator 170 ~ oxygen supply device 180 ~ temperature adjustment device 190 ~ culture solution supplement device c ~ cell Μ ~ metabolic fluid
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