TW201623604A - Culture container - Google Patents
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- TW201623604A TW201623604A TW104131756A TW104131756A TW201623604A TW 201623604 A TW201623604 A TW 201623604A TW 104131756 A TW104131756 A TW 104131756A TW 104131756 A TW104131756 A TW 104131756A TW 201623604 A TW201623604 A TW 201623604A
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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Abstract
Description
本發明係關於一種培養容器。更具體而言,本發明係關於一種具有抗彎液面效果之培養容器。本案係基於2014年9月25日於日本申請之日本專利特願2014-195164號並主張優先權,並將其內容引用於本文中。 The present invention relates to a culture vessel. More specifically, the present invention relates to a culture vessel having a meniscus resistant effect. The present application is based on Japanese Patent Application No. 2014-195164, filed on Jan.
當加入培養溶液至培養容器內之情形時,因溶液之界面張力之作用而於界面產生凹形之彎液面。尤其於培養時使用凝膠之情況,由於凝膠化前之溶液之黏性較高而溶液易附著於培養容器之內壁面,因此該傾向變得更明顯。於觀察培養面時,在形成彎液面的部分形成陰影,因此妨礙存在於該部分之培養細胞之觀察。 When the culture solution is added to the culture vessel, a concave meniscus is generated at the interface due to the interfacial tension of the solution. In particular, in the case where a gel is used for the culture, the tendency becomes more conspicuous because the viscosity of the solution before gelation is high and the solution tends to adhere to the inner wall surface of the culture vessel. When the culture surface is observed, a shadow is formed in the portion where the meniscus is formed, thereby obstructing the observation of the cultured cells existing in the portion.
作為用以解決彎液面之問題之技術,於日本特開2001-340070號公報(專利文獻1)中揭示有一種細胞培養器,其特徵在於:於培養容器底面具有層A且於層A上重疊地具有層B,該層A含有來自活體之物質,且因加溫而變成凝膠狀態之溶液以未凝膠化狀態冷凍而成,該層B係由不會因加溫而變成凝膠狀態之水溶液冷凍而成。 As a technique for solving the problem of the meniscus, a cell culture device is disclosed in Japanese Laid-Open Patent Publication No. 2001-340070 (Patent Document 1), which has a layer A on the bottom surface of the culture container and on the layer A. Superposedly having a layer B containing a substance derived from a living body, and a solution which becomes a gel state by heating is frozen in an ungelled state, and the layer B is formed into a gel by heating. The aqueous solution of the state is frozen.
[專利文獻1]日本特開2001-340070號公報 [Patent Document 1] Japanese Patent Laid-Open Publication No. 2001-340070
習知之用以解決彎液面之問題的技術係藉由「發揮降低或防止彎液面之產生本身的作用」此手段而完成。 The conventional technique for solving the problem of the meniscus is accomplished by "acting to reduce or prevent the occurrence of the meniscus itself".
另一方面,本發明之目的在於提供一種允許彎液面之產生,並且可解決彎液面之問題的培養容器。 On the other hand, an object of the present invention is to provide a culture container which allows the generation of a meniscus and which solves the problem of the meniscus.
本發明人進行潛心研究,結果發現藉由使培養容器具有「將彎液面設置在應培養及觀察之範圍之外側」的構造,可達成上述本發明之目的,從而完成了本發明。 As a result of intensive studies, the inventors of the present invention have achieved the object of the present invention by providing a structure in which the culture vessel has "the meniscus is disposed outside the range to be cultured and observed", and completed the present invention.
(1) (1)
本發明之培養容器含有培養室,該培養室具有底面、及設置於該底面之周緣的周側壁面。於培養室內,沿著底面之周緣環繞設置有障壁面。障壁面係以如下方式形成:使底面上之較周緣更內側之內部周緣作為基礎端上升,朝向周側壁面,且形成與周側壁面之邊界。 The culture container of the present invention comprises a culture chamber having a bottom surface and a circumferential side wall surface provided on a periphery of the bottom surface. In the culture chamber, a barrier surface is provided around the circumference of the bottom surface. The barrier surface is formed such that the inner peripheral edge of the bottom surface on the inner side of the outer periphery rises as a base end, faces the peripheral side wall surface, and forms a boundary with the peripheral side wall surface.
藉由以於在較周緣更內側而隔開之位置存在內部周緣之方式構成,而至少降低當培養室內收容有培養液時產生之彎液面的影響。障壁面被設定在可形成如下程度差距的高度:抑制障壁面上之細胞殘留之程度。 The inner peripheral edge is formed at a position spaced apart from the inner side of the circumference to at least reduce the influence of the meniscus generated when the culture liquid is accommodated in the culture chamber. The barrier surface is set at a height that can form a degree of difference: the degree of cell residue on the barrier surface is suppressed.
根據上述構成,可減少於因培養液面產生彎液面而變得難以觀察之培 養室內的周緣部分存在培養細胞的情況。因此,儘管允許產生彎液面,但可擴大已培養之細胞的觀察範圍。 According to the above configuration, it is possible to reduce the cultivation which is difficult to observe due to the occurrence of the meniscus on the culture liquid surface. The cultured cells are present in the peripheral part of the nursery. Therefore, although the meniscus is allowed to be produced, the observation range of the cultured cells can be expanded.
(2) (2)
亦可為,自橫向剖視時障壁面之形狀為直線、曲線、及直線與曲線之組合的任一種。 Alternatively, the shape of the barrier surface may be a straight line, a curved line, or a combination of a straight line and a curved line from a transverse cross-sectional view.
如此,允許各種形狀之障壁面變寬。 In this way, the barrier faces of various shapes are allowed to widen.
(3) (3)
較佳為,自橫向剖視時,障壁面係由自內部周緣上升之下側障壁面部、及與該下部障壁面部連接設置且形成與周側壁面之邊界的上側障壁面部構成,自內部周緣上升之下側障壁面部與底面成90度以上且150度以下之角度。 Preferably, since the transverse cross-sectional view, the barrier surface is formed by a lower barrier surface portion that rises from the inner peripheral edge and an upper barrier surface portion that is connected to the lower barrier surface portion and forms a boundary with the peripheral sidewall surface, and rises from the inner peripheral edge. The lower side barrier surface and the bottom surface are at an angle of 90 degrees or more and 150 degrees or less.
藉由下側障壁面部之該角度為90度以上,可不遮擋培養室內之除周緣部分以外之部分的視野而擴大集中之培養細胞之觀察範圍,並且藉由為150度以下,而可進一步減少培養細胞進入難以觀察到的培養室內之周緣部分。 When the angle of the lower barrier surface portion is 90 degrees or more, the observation range of the concentrated cultured cells can be expanded without obstructing the field of view of the portion other than the peripheral portion in the culture chamber, and the culture can be further reduced by 150 degrees or less. The cells enter the peripheral portion of the culture chamber that is difficult to observe.
(4) (4)
於上述(3)之培養容器中,亦可為,自橫向剖視時,將彼此為相同斜率之下側障壁面部與上側障壁面部無邊界地連接設置。 In the culture container according to the above (3), the side barrier portion and the upper barrier portion may be connected to each other without a boundary from the same slope in the transverse cross-sectional view.
於此情形時,障壁面之構造簡單,就製造上之觀點而言亦較佳。 In this case, the structure of the barrier surface is simple, and it is also preferable from the viewpoint of manufacturing.
(5) (5)
當自橫向剖視時障壁面之形狀為曲線時,該曲線較佳為如下形狀:切 線與上述底面所成之角度自底面側朝向周側壁面側變大之形狀。即,亦可為如下形狀:自橫向(相對於底面平行之方向)剖視時障壁面之形狀為曲線,該曲線為曲線之切線與底面所成之角度自底面側朝向周側壁面側逐漸變大之形狀。 When the shape of the barrier surface is a curve from the transverse cross section, the curve preferably has the following shape: The angle formed by the line from the bottom surface is increased from the bottom surface side toward the peripheral side wall surface side. That is, it may be a shape in which the shape of the barrier surface is a curve when viewed from the lateral direction (the direction parallel to the bottom surface), and the curve is such that the angle formed by the tangent to the bottom surface gradually changes from the bottom surface side toward the peripheral side wall surface side. Big shape.
於此情形時,較佳地確保培養細胞不存在於難以觀察到的培養室內之周緣部分,並且就製造上之觀點而言亦較佳。 In this case, it is preferable to ensure that the cultured cells are not present in the peripheral portion of the culture chamber which is difficult to observe, and it is also preferable from the viewpoint of production.
(6) (6)
周側壁面及內部周椽之俯視形狀可為圓,於此情形時,內部周緣之直徑可為30mm以上且170mm以下。 The shape of the circumferential side wall surface and the inner circumference of the circumference may be a circle. In this case, the diameter of the inner circumference may be 30 mm or more and 170 mm or less.
藉由內部周緣之直徑為30mm以上,而可較佳地獲取用於培養及觀察之底面上之空間,且藉由為170mm以下,而可較佳地享有將已培養之細胞大部分集中於可觀察到之部分的效果。 By having a diameter of 30 mm or more in the inner circumference, it is preferable to obtain a space on the bottom surface for cultivation and observation, and by being 170 mm or less, it is preferable to concentrate most of the cultured cells. The effect of the observed part.
根據本發明,可提供一種即便產生彎液面,因該彎液面而形成之陰影亦不會妨礙培養細胞之觀察的培養容器。 According to the present invention, it is possible to provide a culture container in which the shadow formed by the meniscus does not hinder the observation of the cultured cells even if a meniscus is generated.
100、1001‧‧‧培養容器 100, 1001‧‧‧ culture container
200‧‧‧底面 200‧‧‧ bottom
300、300b、300d‧‧‧周側壁面 300, 300b, 300d‧‧‧ weeks side wall
400、400a、400b、400c、400d、400e、400f、400g、400h、400i、400j、400k‧‧‧障壁面 400, 400a, 400b, 400c, 400d, 400e, 400f, 400g, 400h, 400i, 400j, 400k‧ ‧ barrier
410、410f、410g、410h、410j、410k‧‧‧下側障壁面部 410, 410f, 410g, 410h, 410j, 410k‧‧‧ lower barrier face
420、420d、420e、420f、420g、420h、420j、420k‧‧‧上側障壁面部 420, 420d, 420e, 420f, 420g, 420h, 420j, 420k‧‧‧ upper barrier face
O、B‧‧‧(底面之)周緣 O, B‧‧‧ (bottom) circumference
B2、B2a‧‧‧(底面之)內部周緣 B2, B2a‧‧‧ (bottom) internal circumference
B3、B3b、B3d‧‧‧(障壁面與周側壁面之)邊界 B3, B3b, B3d‧‧‧ (Bound wall and circumferential side wall) boundary
C‧‧‧培養室 C‧‧‧Cultivation room
θ 1‧‧‧(下側障壁面部與底面所成之)角度 θ 1‧‧‧ (the lower barrier face and the bottom surface)
θ、θ'、θ"‧‧‧(障壁面之剖面形狀之切線與底面所成之)角度 θ, θ', θ"‧‧‧ (the tangent to the bottom of the cross-sectional shape of the barrier surface)
圖1表示第1實施形態之培養容器之示意性外觀立體圖。 Fig. 1 is a schematic external perspective view of a culture container according to a first embodiment.
圖2表示第1實施形態之培養容器使用時的示意性剖面圖。 Fig. 2 is a schematic cross-sectional view showing the use of the culture container of the first embodiment.
圖3表示圖2之示意性剖面圖之局部放大圖。 Fig. 3 is a partial enlarged view of the schematic sectional view of Fig. 2;
圖4表示圖2之示意性剖面圖之要部放大圖。 Fig. 4 is an enlarged view of an essential part of the schematic sectional view of Fig. 2;
圖5表示第1實施形態之第1變形例之培養容器使用時的示意性剖面圖。 Fig. 5 is a schematic cross-sectional view showing the use of the culture container according to the first modification of the first embodiment.
圖6表示第1實施形態之第2變形例之培養容器使用時的示意性剖面圖。 Fig. 6 is a schematic cross-sectional view showing the use of the culture container according to the second modification of the first embodiment.
圖7表示第1實施形態之第3變形例之培養容器使用時的示意性剖面圖。 Fig. 7 is a schematic cross-sectional view showing the use of the culture container according to the third modification of the first embodiment.
圖8表示第1實施形態之第4變形例之培養容器使用時的示意性剖面圖。 Fig. 8 is a schematic cross-sectional view showing the use of the culture container according to the fourth modification of the first embodiment.
圖9表示第1實施形態之第5變形例之培養容器使用時的示意性剖面圖。 Fig. 9 is a schematic cross-sectional view showing the use of the culture container according to the fifth modification of the first embodiment.
圖10表示第2實施形態之培養容器之要部放大圖。 Fig. 10 is an enlarged view of a main part of a culture container according to a second embodiment.
圖11表示第3實施形態之培養容器之要部放大圖。 Fig. 11 is an enlarged view of a main part of a culture container according to a third embodiment.
圖12表示第4實施形態之培養容器之要部放大圖。 Fig. 12 is an enlarged view of a main part of a culture container according to a fourth embodiment.
圖13表示第5實施形態之培養容器之要部放大圖。 Fig. 13 is an enlarged view of a main part of a culture container according to a fifth embodiment.
圖14表示第6實施形態之培養容器之要部放大圖。 Fig. 14 is an enlarged view of a main part of the culture container according to the sixth embodiment.
圖15表示第7實施形態之培養容器之要部放大圖。 Fig. 15 is an enlarged view of a main part of a culture container according to a seventh embodiment.
圖16表示第8實施形態之培養容器之示意性外觀立體圖。 Fig. 16 is a schematic perspective view showing the culture container of the eighth embodiment.
圖17係於實施例1中製成之培養容器加入培養基後上下方向觀察之照片。 Fig. 17 is a photograph of the culture container prepared in Example 1 after being added to the culture medium, and then viewed from above and below.
圖18係於實施例1中製成之培養容器內進行細胞培養後上下方向觀察之照片(局部放大)。 Fig. 18 is a photograph (partial enlargement) of the cell culture after the cell culture was carried out in the culture vessel prepared in Example 1.
圖19係於實施例1中製成之培養容器內進行細胞培養後上下方向觀察 之照片(局部放大)。 Figure 19 is a view of the up-and-down direction after cell culture in the culture vessel prepared in Example 1. Photo (partial enlargement).
圖20係於實施例2中製成之培養容器加入培養基後上下方向觀察之照片。 Fig. 20 is a photograph of the culture container prepared in Example 2, which was observed in the vertical direction after being added to the culture medium.
圖21係於實施例2中製成之培養容器內進行細胞培養後之下方向觀察之照片(局部放大)。 Fig. 21 is a photograph (partial enlargement) of the observation in the lower direction after cell culture in the culture vessel prepared in Example 2.
圖22係於實施例2中製成之培養容器內進行細胞培養後上下方向觀察之照片(局部放大)。 Fig. 22 is a photograph (partial enlargement) of the cell culture in the culture vessel prepared in Example 2, which was observed in the vertical direction.
圖23表示比較例1中製成之培養容器之示意性剖面圖。 Fig. 23 is a schematic cross-sectional view showing a culture vessel prepared in Comparative Example 1.
圖24係於比較例1中製成之培養容器中加入培養基後上下方向觀察之照片。 Fig. 24 is a photograph of the culture container prepared in Comparative Example 1 which was observed in the vertical direction after the medium was added.
圖25係於比較例1中製成之培養容器內進行細胞培養後上下方向觀察之照片(局部放大)。 Fig. 25 is a photograph (partial enlargement) of the cell culture after the cell culture was carried out in the culture vessel prepared in Comparative Example 1.
圖26係於比較例1中製成之培養容器內進行細胞培養後上下方向觀察之照片(局部放大)。 Fig. 26 is a photograph (partial enlargement) of the cells cultured in the culture vessel prepared in Comparative Example 1 and observed in the vertical direction.
以下,一面參照圖式,一面對本發明之實施形態進行說明。於以下之說明中,對相同之要素標註相同之符號,只要未作特別說明,則其等之名稱及功能亦相同。因此,基本上不重複對其等之詳細說明。 Hereinafter, embodiments of the present invention will be described with reference to the drawings. In the following description, the same elements are denoted by the same reference numerals, and the names and functions thereof are the same unless otherwise specified. Therefore, the detailed description thereof will not be repeated.
[第1實施形態] [First Embodiment]
圖1表示第1實施形態之培養容器之示意性外觀立體圖。圖2表示第1實施形態之培養容器之使用時示意性剖面圖。圖2之示意性剖面圖係自橫 向觀察以包含圖1之A-A線之鉛垂面切斷之面之圖。圖3表示圖2之示意性剖面圖之局部放大。圖4表示圖2之示意性剖面圖之要部(圓圈部分)之放大圖。再者,為了方便說明,將各圖式之上側稱為上,將下側稱為下。 Fig. 1 is a schematic external perspective view of a culture container according to a first embodiment. Fig. 2 is a schematic cross-sectional view showing the use of the culture container of the first embodiment. Figure 2 is a schematic cross-sectional view of the horizontal The view of the surface cut by the vertical plane including the line A-A of Fig. 1 was observed. Fig. 3 is a partial enlarged view of the schematic sectional view of Fig. 2. Fig. 4 is an enlarged view showing an essential part (circular portion) of the schematic sectional view of Fig. 2. In addition, for convenience of description, the upper side of each drawing is called upper, and the lower side is called lower.
如圖1及圖2所示,培養容器100含有底面200、周側壁面300及障壁面400作為構成其內側之面。由該等底面200、周側壁面300及障壁面400界定培養室C。換言之,本實施形態之培養容器含有培養室C,該培養室C具備底面200、沿著底面200之周緣B2設置於外側之障壁面400、及沿著障壁面400之周緣設置於更外側之周側壁面300。 As shown in FIGS. 1 and 2, the culture container 100 includes a bottom surface 200, a peripheral side wall surface 300, and a barrier wall surface 400 as surfaces constituting the inner side thereof. The culture chamber C is defined by the bottom surface 200, the peripheral side wall surface 300, and the barrier surface 400. In other words, the culture container according to the present embodiment includes the culture chamber C including the bottom surface 200, the barrier wall surface 400 provided outside the peripheral edge B2 of the bottom surface 200, and the circumference provided on the outer side along the periphery of the barrier surface 400. Side wall surface 300.
底面200於上下方向觀察為圓形,且如圖3所示,設置有自該底面200之周緣O之全周上升且上方開口之周側壁面300。沿著底面200之周緣O之全周以特定高度h環繞設置之障壁面400係以如下方式形成:使自底面200之周緣O向內側隔開特定距離w之內部周緣B2作為基礎端而上升,朝向周側壁面300,且於特定高度h之位置形成與周側壁面300之邊界B3。 The bottom surface 200 is circular in the vertical direction, and as shown in FIG. 3, a peripheral side wall surface 300 which is raised from the entire circumference O of the bottom surface 200 and which is opened upward is provided. The barrier rib 400 disposed around the entire circumference of the peripheral edge O of the bottom surface 200 at a specific height h is formed by raising the inner periphery B2 of the specific distance w from the periphery O of the bottom surface 200 as a base end. The peripheral side wall surface 300 is formed, and a boundary B3 with the peripheral side wall surface 300 is formed at a position of a specific height h.
如圖3所示,特定距離w相當於周緣O與內部周緣B2之距離。特定高度h相當於周緣O與邊界B3之距離,且設定為可構成藉由與特定距離w之關係而抑制細胞殘留於障壁面400之形狀。於本實施形態中設定為:成為應收容於培養室C內之培養液之最低水位H以下。 As shown in FIG. 3, the specific distance w corresponds to the distance between the circumference O and the inner circumference B2. The specific height h corresponds to the distance between the peripheral edge O and the boundary B3, and is set so as to prevent the cell from remaining in the shape of the barrier rib 400 by the relationship with the specific distance w. In the present embodiment, it is set to be equal to or lower than the lowest water level H of the culture solution to be contained in the culture chamber C.
於培養液面S產生彎液面M之情形時,培養室C之周緣部分之培養液面S因彎液面M現象而成為曲面狀,因此自上下方向觀察之視野於該部分變得難以觀察。於培養容器100,藉由設置自內部周緣B2上升之特定高度h之障壁面400,而阻止培養細胞自內部周緣B2向障壁面400 側移動,藉由使障壁面400自內部周緣B2朝向外方向且使內部周緣B2與周側壁面300隔開特定距離w,而使產生彎液面M之部分之至少一部分設置在內部周緣B2外。因此,儘管允許彎液面M之產生,但可使已培養之細胞的大部分存在於可觀察到之內部周緣B2內部。 When the meniscus surface M is generated in the culture liquid surface S, the culture liquid surface S of the peripheral portion of the culture chamber C is curved due to the meniscus M phenomenon, so that the field of view viewed from the up and down direction becomes difficult to observe in this portion. . In the culture vessel 100, the cultured cells are prevented from moving from the inner circumference B2 to the barrier surface 400 by providing the barrier surface 400 at a specific height h rising from the inner periphery B2. The side movement is such that at least a part of the portion where the meniscus M is generated is disposed outside the inner circumference B2 by causing the barrier surface 400 to face outward from the inner circumference B2 and the inner circumference B2 and the peripheral side wall surface 300 by a specific distance w. . Therefore, although the meniscus M is allowed to be generated, most of the cultured cells can be present inside the inner periphery B2 which can be observed.
藉由障壁面400具有相當於特定距離w之寬度,而至少降低於培養室C內收容有培養液時產生於培養液面S之彎液面M的影響。因此,該特定距離w並無特別限定。例如,存在特定距離w越大,越能進一步降低彎液面M之影響之傾向,並且存在特定距離w越小,越可充分獲取用於培養及觀察之底面200上之空間之傾向,業者可基於該等兩種傾向而適當決定。作為具體例,特定距離w可為2mm以上且3mm以下。於本實施形態中,就避免彎液面M之影響之觀點而言,特定距離w被設定為彎液面M之寬度W以上。 The barrier surface 400 has a width corresponding to the specific distance w, and at least reduces the influence of the meniscus M generated on the culture liquid surface S when the culture liquid is contained in the culture chamber C. Therefore, the specific distance w is not particularly limited. For example, the larger the specific distance w is, the more the tendency of the meniscus M is further reduced, and the smaller the specific distance w, the more the space for the culture and observation of the bottom surface 200 is sufficiently obtained. It is appropriately determined based on these two tendencies. As a specific example, the specific distance w may be 2 mm or more and 3 mm or less. In the present embodiment, the specific distance w is set to be equal to or larger than the width W of the meniscus M from the viewpoint of avoiding the influence of the meniscus M.
特定高度h只要設定為可構成如下差距即可,因此並無特別限定,該差距係藉由與特定距離w之關係而抑制細胞殘留於障壁面400之差距。例如,特定高度h為1mm以上且21mm以下,較佳為1mm以上且15mm以下。藉由為上述下限值以上,而可更為減少培養細胞滲入上下方向觀察時難以觀察到之內部周緣B2之外側部分,並且藉由為上述上限值以下,而可充分獲取應收容培養液之容積。於本實施形態中,就更充分地獲取應收容培養液之容積之觀點而言,特定高度h被設定在應收容於培養室C內之培養液之水位H以下。於此情形時,培養液面S與培養容器100之內壁面之邊界E處於周側壁面300上。 The specific height h is not particularly limited as long as it can be set as follows, and the difference is such that the difference in cell retention between the barrier faces 400 is suppressed by the relationship with the specific distance w. For example, the specific height h is 1 mm or more and 21 mm or less, preferably 1 mm or more and 15 mm or less. By setting it to the above lower limit value, it is possible to further reduce the outer portion of the inner peripheral edge B2 which is difficult to be observed when the cultured cells are infiltrated in the vertical direction, and it is possible to sufficiently obtain the culture liquid to be accommodated by being equal to or less than the above upper limit value. Volume. In the present embodiment, the specific height h is set to be equal to or lower than the water level H of the culture solution to be accommodated in the culture chamber C from the viewpoint of more sufficiently acquiring the volume of the culture solution to be stored. In this case, the boundary E between the culture liquid surface S and the inner wall surface of the culture vessel 100 is on the peripheral side wall surface 300.
如圖4所示,障壁面400由下側障壁面部410及上側障壁面 部420構成。下側障壁面部410係以與底面200成角度θ 1之方式將內部周緣B2作為基礎端上升,並於高度h1之位置與上側障壁面部420連接設置。於高度h1之位置,下側障壁面部410與上側障壁面部420之邊界形成脊線。上側障壁面部420相對於底面200呈角度θ 2地向周側壁面300傾斜,並且與周側壁面300形成邊界B3。 As shown in FIG. 4, the barrier surface 400 is composed of a lower barrier surface portion 410 and an upper barrier surface. The unit 420 is configured. The lower barrier surface portion 410 is raised from the base end B2 so as to be at an angle θ 1 to the bottom surface 200, and is connected to the upper barrier faucet portion 420 at a position of the height h1. At the position of the height h1, a ridge line is formed at the boundary between the lower barrier portion 410 and the upper barrier portion 420. The upper barrier façade 420 is inclined toward the peripheral side wall surface 300 at an angle θ 2 with respect to the bottom surface 200, and forms a boundary B3 with the circumferential side wall surface 300.
下側障壁面部410與底面200形成之角度θ 1為90度。藉此,可有效地防止底面200中之培養細胞超過內部周緣B2而向外側移動。作為上側障壁面部420之傾斜角之角度θ 2並無特別限定。例如存在如下傾向:角度θ 2越大,越可充分確保下側障壁面部410之高度h1,而更為減少培養細胞滲入“上下方向觀察時難以觀察到之內部周緣B2之外側部分”之傾向;角度θ 2越小,越可充分獲取應收容培養液之容積之傾向;業者可基於該等兩種傾向而適當決定。 The angle θ 1 formed by the lower barrier surface portion 410 and the bottom surface 200 is 90 degrees. Thereby, it is possible to effectively prevent the cultured cells in the bottom surface 200 from moving outward beyond the inner periphery B2. The angle θ 2 of the inclination angle of the upper barrier surface portion 420 is not particularly limited. For example, there is a tendency that the larger the angle θ 2 is, the more the height h1 of the lower barrier surface portion 410 can be sufficiently ensured, and the tendency of the cultured cells to infiltrate into the outer portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up-and-down direction is further reduced; The smaller the angle θ 2 is, the more the volume of the culture solution to be accommodated can be sufficiently obtained; the operator can appropriately determine the two tendencies based on the two tendencies.
下側障壁面部410之高度h1並無特別限定。例如存在如下傾向:高度h1越大,則更加減少“培養細胞滲入上下方向觀察時難以觀察到之內部周緣B2之外側部分”之傾向;高度h1越小,越可充分獲取應收容培養液之容積之傾向,業者可基於該等兩種傾向而適當決定。具體而言,高度h1例如可設定為0.5mm,亦可設定為1mm。 The height h1 of the lower barrier surface portion 410 is not particularly limited. For example, there is a tendency that the larger the height h1 is, the more the "the outer portion of the inner peripheral edge B2 which is difficult to be observed when the cultured cells are infiltrated in the vertical direction" is further reduced; the smaller the height h1, the more the volume of the culture medium to be accommodated can be sufficiently obtained. The tendency of the industry can be appropriately determined based on these two tendencies. Specifically, the height h1 can be set, for example, to 0.5 mm, or can be set to 1 mm.
內部周緣B2之直徑可與習知之培養容器之內底面之直徑相同。例如為30mm以上且170mm以下,較佳為32mm以上且90mm以下。藉由為上述下限值以上,而可較佳地獲取用以培養及觀察之底面200上之空間,並且藉由為上述上限值以下,而可較佳地享有將已培養之細胞之大部分集中於可觀察到之部分的效果。 Diameter of the inner circumference B2 It can be the same diameter as the inner bottom surface of the conventional culture container. For example, it is 30 mm or more and 170 mm or less, preferably 32 mm or more and 90 mm or less. By being at least the above lower limit value, it is possible to preferably obtain a space on the bottom surface 200 for cultivation and observation, and by being equal to or less than the above upper limit value, it is preferable to enjoy the large size of the cultured cells. Partly focused on the effects of the observable parts.
培養容器100可由具有透明性之原材料構成。作為具有透明性之原材料,可列舉以玻璃及石英為代表之無機物、以及以合成樹脂為代表之有機物。作為合成樹脂,可列舉聚苯乙烯(PS)、聚丙烯(PP)、聚甲基戊烯(PMP)、聚碳酸酯(PC)、聚甲基丙烯酸甲酯(PMMA)、聚甲基丙烯酸甲醯亞胺(PMMI)、及環烯共聚物(COC)等聚合物。進而,亦可列舉由該等聚合物中2個以上之單體單元合成之共聚物。於為合成樹脂之情形時,就容易成形且不易破裂之方面而言較佳。 The culture container 100 can be composed of a raw material having transparency. Examples of the transparent material include inorganic materials represented by glass and quartz, and organic materials represented by synthetic resins. Examples of the synthetic resin include polystyrene (PS), polypropylene (PP), polymethylpentene (PMP), polycarbonate (PC), polymethyl methacrylate (PMMA), and polymethyl methacrylate. Polymers such as quinone imine (PMMI) and cycloolefin copolymer (COC). Further, a copolymer synthesized from two or more monomer units of the polymers may also be mentioned. In the case of a synthetic resin, it is preferable in terms of being easily formed and not easily broken.
培養容器100可為一體成形之構造體,亦可為由例如構成底面200之構件與構成周側壁面300及障壁面400之構件之組合而形成之構造體。為由構件之組合而形成之構造體時,各個構件亦可由不同之原材料構成。 The culture container 100 may be an integrally formed structure, or may be a structure formed by, for example, a combination of a member constituting the bottom surface 200 and members constituting the peripheral side wall surface 300 and the barrier wall surface 400. In the case of a structure formed by a combination of members, each member may be composed of different raw materials.
就物理性、化學性及/或生物化學觀點而言,亦可對培養容器100之底面200、周側壁面300及障壁面400實施表面處理。只要為業者,則可適當選擇此種表面處理。例如,亦可對障壁面400、或對障壁面400及周側壁面300實施用以妨礙細胞培養之表面處理。 The bottom surface 200, the peripheral side wall surface 300, and the barrier wall surface 400 of the culture vessel 100 may also be subjected to a surface treatment in terms of physical, chemical, and/or biochemical viewpoints. As long as it is a manufacturer, such a surface treatment can be appropriately selected. For example, the barrier surface 400, or the barrier surface 400 and the peripheral sidewall surface 300 may be subjected to a surface treatment for hindering cell culture.
進而,亦可根據培養容器100之使用目的等,設置未圖示之更多附加構造。只要為業者,則可適當選擇此種附加構造。具體而言,可列舉用以使培養室C成為加蓋狀態之蓋體、用以間隔內容物之間隔件及用以特定出內容物(尤其為細胞群)之量及位置之網格圖案等設置於底面200之引導構造。 Further, a further additional structure (not shown) may be provided depending on the purpose of use of the culture container 100 or the like. As long as it is a manufacturer, such an additional structure can be appropriately selected. Specifically, a lid body for making the culture chamber C into a capped state, a spacer for spacing the contents, and a grid pattern for specifying the amount and position of the contents (particularly, a cell population) may be mentioned. A guiding structure provided on the bottom surface 200.
<變形例> <Modification>
培養容器100除已圖示者以外還被允許有各種變形例。於變形例中, 只要彎液面M之至少一部分設置在較內部周緣B2更外側即可,因此只要邊界E不形成於障壁面400中下側障壁面410上,則可允許為任何構造,該下側障壁面410為將內部周緣B2作為基礎端並相對於底面200呈90度上升者。即,於變形例中,當已收容培養液至培養室C內時,只要以培養液之液面S與培養容器100之內壁之邊界E成為較內部周緣B2更外側之方式構成即可。 The culture container 100 is allowed to have various modifications in addition to those already illustrated. In the variant, As long as at least a part of the meniscus M is disposed outside the inner peripheral edge B2, any configuration may be allowed as long as the boundary E is not formed on the lower barrier surface 410 in the barrier face 400, and the lower barrier face 410 may be allowed. The inner circumference B2 is used as a base end and is raised at 90 degrees with respect to the bottom surface 200. In other words, in the modified example, when the culture liquid has been stored in the culture chamber C, the boundary E between the liquid surface S of the culture liquid and the inner wall of the culture container 100 may be configured to be outside the inner circumference B2.
以下,列舉圖5至圖9,對特定距離w及特定高度h之組合不同的第1實施形態之變形例進行說明。圖5至圖9對應於第1實施形態之圖3。於以下之變形例中,主要對與第1實施形態不同之方面進行說明,省略相同方面之說明。 Hereinafter, a modification of the first embodiment in which the combination of the specific distance w and the specific height h is different will be described with reference to FIGS. 5 to 9. 5 to 9 correspond to Fig. 3 of the first embodiment. In the following modifications, the differences from the first embodiment will be mainly described, and the description of the same aspects will be omitted.
[第1變形例] [First Modification]
圖5表示第1實施形態之第1變形例。圖5所示之培養容器100a之障壁面400a,係藉由將特定距離wa設定為短於第1實施形態之特定距離w,而使基礎端即內部周緣B2a處於較第1實施形態之內部周緣B2更外側。藉此,設置在較內部周緣B2a更外側之彎液面M成為該障壁面400a之一部分,可降低因彎液面M所致之影響。即,於本變形例中,因彎液面M設置在較內部周緣B2a更外側,因此亦可降低因彎液面M所致之影響。 Fig. 5 shows a first modification of the first embodiment. The barrier surface 400a of the culture container 100a shown in Fig. 5 is set to be shorter than the specific distance w of the first embodiment by setting the specific distance wa to be shorter than the inner circumference B2a of the first embodiment. B2 is more lateral. Thereby, the meniscus M provided outside the inner peripheral edge B2a becomes a part of the barrier face 400a, and the influence by the meniscus M can be reduced. That is, in the present modification, since the meniscus M is disposed outside the inner peripheral edge B2a, the influence due to the meniscus M can be reduced.
[第2變形例] [Second Modification]
圖6表示第1實施形態之第2變形例。圖6所示之培養容器100b之障壁面400b係藉由將特定高度hb設定為大於第1實施形態之特定高度h,而使與周側壁面300b之邊界B3b處於較第1實施形態之邊界B3更上側。藉此,可更為減少培養細胞滲入“上下方向觀察時難以觀察到之內部周緣B2 之外側部分”。於本變形例中,特定高度hb成為應收容於培養室C內之培養液之水位H以上,但未達培養液面S與周側壁面300b之邊界E之高度。 Fig. 6 shows a second modification of the first embodiment. The barrier surface 400b of the culture container 100b shown in Fig. 6 is set to be larger than the specific height h of the first embodiment by the specific height hb, so that the boundary B3b with the peripheral side wall surface 300b is at the boundary B3 of the first embodiment. More on the side. Thereby, the culture cells can be further infiltrated into the inner periphery B2 which is difficult to observe when viewed from the top and bottom direction. In the present modification, the specific height hb is equal to or higher than the water level H of the culture solution to be accommodated in the culture chamber C, but does not reach the height E of the boundary E between the culture liquid surface S and the circumferential side wall surface 300b.
[第3變形例] [Third Modification]
圖7表示第1實施形態之第3變形例。圖7所示之培養容器100c之障壁面400c與第1變形例同樣地,係藉由將特定距離wa設定為短於第1實施形態之特定距離w,而使基礎端即內部周緣B2a處於較第1實施形態之內部周緣B2更外側。藉此,設置在較內部周緣B2a更外側之彎液面M成為該障壁面400c之一部分,可降低因彎液面M所致之影響。即,於本變形例中,因彎液面M設置在較內部周緣B2a更外側,因此亦可降低因彎液面M所致之影響。 Fig. 7 shows a third modification of the first embodiment. In the same manner as in the first modification, the barrier wall surface 400c of the culture container 100c shown in Fig. 7 is such that the base end, that is, the inner circumference B2a, is set by setting the specific distance wa to be shorter than the specific distance w of the first embodiment. The inner periphery B2 of the first embodiment is further outside. Thereby, the meniscus M provided outside the inner peripheral edge B2a becomes a part of the barrier face 400c, and the influence by the meniscus M can be reduced. That is, in the present modification, since the meniscus M is disposed outside the inner peripheral edge B2a, the influence due to the meniscus M can be reduced.
進而,與第2變形例同樣地,障壁面400c與周側壁面300b之邊界B3b處於較第1實施形態之邊界B3更上側。藉此,可更為減少培養細胞滲入“上下方向觀察時難以觀察到之內部周緣B2a之外側部分”。於本變形例中,特定高度hb成為應收容於培養室C內之培養液之水位H以上,但未達培養液面S與周側壁面300b之邊界E之高度。 Further, similarly to the second modification, the boundary B3b between the barrier rib surface 400c and the peripheral side wall surface 300b is higher than the boundary B3 of the first embodiment. Thereby, it is possible to further reduce the infiltration of the cultured cells into the "outer side portion of the inner peripheral edge B2a which is difficult to observe when viewed in the up-and-down direction". In the present modification, the specific height hb is equal to or higher than the water level H of the culture solution to be accommodated in the culture chamber C, but does not reach the height E of the boundary E between the culture liquid surface S and the circumferential side wall surface 300b.
[第4變形例] [Fourth Modification]
圖8表示第1實施形態之第4變形例。圖8所示之培養容器100d之障壁面400d,係藉由將特定高度hd設定為更大於第2變形例之特定高度hb,而使與周側壁面300d之邊界B3d處於較第2變形例之邊界B3b進而更上側。藉此,可進一步減少培養細胞滲入“上下方向觀察時難以觀察到之內部周緣B2之外側部分”。 Fig. 8 shows a fourth modification of the first embodiment. The barrier surface 400d of the culture container 100d shown in Fig. 8 is set to be larger than the specific height hb of the second modification by the specific height hd, and the boundary B3d with the peripheral side wall surface 300d is in the second modification. The boundary B3b is further on the upper side. Thereby, it is possible to further reduce the infiltration of the cultured cells into the "outer side portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up-and-down direction".
於本變形例中,培養液面S與培養容器100d之內壁面之邊 界Ed處於上側周側壁面部420d上,因此障壁面400d之特定高度hd超過應收容於培養室C內之培養液之水位H且為邊界Ed之高度以上。但是,由於設置有足夠量之特定距離w,因此可使彎液面M設置在內部周緣B2之外側,而可避免彎液面M之影響。 In the present modification, the culture liquid surface S and the inner wall surface of the culture container 100d are Since the boundary Ed is on the upper peripheral side wall surface portion 420d, the specific height hd of the barrier surface 400d exceeds the water level H of the culture liquid to be accommodated in the culture chamber C and is equal to or higher than the height of the boundary Ed. However, since a sufficient amount of the specific distance w is provided, the meniscus M can be disposed on the outer side of the inner circumference B2, and the influence of the meniscus M can be avoided.
[第5變形例] [Fifth Modification]
圖9表示第1實施形態之第5變形例。與第4變形例同樣地,圖9所示之培養容器100e之障壁面400e,係藉由將特定高度hd設定為更大於第2、第3變形例之特定高度hb,而使與周側壁面300d之邊界B3d處於較第2、第3變形例之邊界B3b進而更上側。藉此,可更為減少培養細胞滲入“上下方向觀察時難以觀察到之內部周緣B2a之外側部分”。 Fig. 9 shows a fifth modification of the first embodiment. Similarly to the fourth modification, the barrier surface 400e of the culture container 100e shown in FIG. 9 is formed so as to have a specific height hd larger than the specific height hb of the second and third modifications. The boundary B3d of 300d is at a higher boundary than the boundary B3b of the second and third modifications. Thereby, it is possible to further reduce the infiltration of the cultured cells into the "outer side portion of the inner peripheral edge B2a which is difficult to observe when viewed in the up-and-down direction".
於本變形例中,培養液面S與培養容器100e之內壁面之邊界Ee處於上側周側壁面部420e上,因此障壁面400e之特定高度hd超過應收容於培養室C內之培養液之水位H且為邊界Ee之高度以上。進而,與第1變形例同樣地,藉由將特定距離wa設定為短於第1實施形態之特定距離w,而使基礎端即內部周緣B2a處於較第1實施形態之內部周緣B2更外側。藉此,設置在較內部周緣B2a更外側之彎液面M成為該障壁面400e之一部分,可降低因彎液面M所致之影響。即,於本變形例中,因彎液面M設置在較內部周緣B2a更靠外側,因此亦可降低因彎液面M所致之影響。 In the present modification, the boundary Ee between the culture liquid surface S and the inner wall surface of the culture container 100e is on the upper circumferential side wall surface portion 420e. Therefore, the specific height hd of the barrier surface 400e exceeds the water level H of the culture liquid to be accommodated in the culture chamber C. And it is above the height of the boundary Ee. Further, similarly to the first modification, by setting the specific distance wa to be shorter than the specific distance w of the first embodiment, the inner end B2a which is the base end is located outside the inner periphery B2 of the first embodiment. Thereby, the meniscus M provided on the outer side of the inner peripheral edge B2a becomes a part of the barrier face 400e, and the influence by the meniscus M can be reduced. That is, in the present modification, since the meniscus M is provided outside the inner peripheral edge B2a, the influence by the meniscus M can be reduced.
<其他實施形態> <Other Embodiments>
於本發明中,障壁面400只要具有自橫向剖視時通過內部周緣B2與邊界B3之間之形狀(連接內部周緣B2與邊界B3之形狀)即可,並不限定於第1實施形態所示之形狀。例如,下側障壁面部410與上側障壁面部420 之邊界可如圖4所示般形成脊線,亦可對該邊界進行C倒角加工或R倒角加工。 In the present invention, the barrier surface 400 is not limited to the first embodiment as long as it has a shape passing between the inner peripheral edge B2 and the boundary B3 (a shape connecting the inner peripheral edge B2 and the boundary B3) from the transverse cross-sectional view. The shape. For example, the lower barrier portion 410 and the upper barrier portion 420 The boundary may be formed as shown in FIG. 4, or C-chamfering or R-chamfering may be performed on the boundary.
進而,障壁面400之自橫向觀察之剖面形狀可為直線,亦可為曲線,還可為直線與曲線之組合,廣泛允許為各種形狀。就更確實地使培養細胞不存在於上下方向觀察時難以觀察到之內部周緣B2之外側部分之觀點而言,障壁面400較佳為如下形狀:自橫向剖視時連結內部周緣B2與邊界B3之直線、或存在於該直線上方之形狀。 Further, the cross-sectional shape of the barrier surface 400 from the lateral direction may be a straight line, a curved line, or a combination of a straight line and a curved line, and various shapes are widely allowed. The barrier wall surface 400 preferably has a shape in which the inner peripheral edge B2 and the boundary B3 are joined from a transverse cross-sectional view, from the viewpoint of more reliably preventing the cultured cells from being present in the outer peripheral portion of the inner peripheral edge B2 when viewed from the upper and lower directions. A straight line or a shape that exists above the line.
以下,列舉圖10至圖15對障壁面400之形狀與第1實施形態不同之第2實施形態至第7實施形態進行說明。圖10至圖15對應於第1實施形態之圖4。於以下之實施形態中,主要對與第1實施形態之不同點進行說明,省略有關相同點之說明。 Hereinafter, the second embodiment to the seventh embodiment in which the shape of the barrier surface 400 is different from that of the first embodiment will be described with reference to Figs. 10 to 15 . 10 to 15 correspond to Fig. 4 of the first embodiment. In the following embodiments, differences from the first embodiment will be mainly described, and the description of the same points will be omitted.
[第2實施形態] [Second Embodiment]
圖10表示第2實施形態之培養容器之要部放大圖。圖10所示之障壁面400f係由下側障壁面部410f及上側障壁面部420f構成。下側障壁面部410f以與底面200呈角度θ 1(90度)之方式將內部周緣B2作為基礎端上升,並與和底面200平行(角度θ 2為180度)之上側障壁面部420f連接設置。再者,圖10中,下側障壁面部410f與上側障壁面部420f之邊界形成脊線,但亦可對該邊界進行C倒角加工或R倒角加工。 Fig. 10 is an enlarged view of a main part of a culture container according to a second embodiment. The barrier surface 400f shown in FIG. 10 is composed of a lower barrier surface portion 410f and an upper barrier surface portion 420f. The lower barrier surface portion 410f is raised from the base edge B2 at an angle θ 1 (90 degrees) to the bottom surface 200, and is connected to the upper barrier faucet portion 420f in parallel with the bottom surface 200 (the angle θ 2 is 180 degrees). Further, in FIG. 10, the ridge line is formed at the boundary between the lower barrier surface portion 410f and the upper barrier surface portion 420f, but the boundary may be subjected to C chamfering or R chamfering.
[第3實施形態] [Third embodiment]
圖11表示第3實施形態之培養容器之要部放大圖。圖11所示之障壁面400g係由下側障壁面部410g及上側障壁面部420g構成。下側障壁面部410g以與底面200呈角度θ 1之方式將內部周緣B2作為基礎端上升,並與和底 面200平行(角度θ 2為180度)之上側障壁面部420g連接設置。 Fig. 11 is an enlarged view of a main part of a culture container according to a third embodiment. The barrier surface 400g shown in FIG. 11 is composed of a lower barrier surface portion 410g and an upper barrier surface portion 420g. The lower barrier surface portion 410g raises the inner peripheral edge B2 as a base end at an angle θ 1 to the bottom surface 200, and the bottom and bottom The upper surface barrier faces 420g are connected in parallel with each other (the angle θ 2 is 180 degrees).
於本實施形態中,角度θ 1可超過90度且為150度以下。藉由超過該下限值,而自上下方向觀察時,可視野良好地觀察應培養及觀察之內部周緣B2內之整個區域,藉由為該上限值以下,而可減少培養細胞進入“上下方向觀察時難以觀察到之內部周緣B2之外側部分”。再者,圖11中,下側障壁面部410g與上側障壁面部420g之邊界形成脊線,但亦可對該邊界進行C倒角加工或R倒角加工。 In the present embodiment, the angle θ 1 may exceed 90 degrees and be 150 degrees or less. When the upper limit is exceeded, the entire region in the inner periphery B2 to be cultured and observed can be observed with good visibility, and the culture cells can be reduced by "upper and lower limits". It is difficult to observe the outer portion of the inner circumference B2 when viewed in the direction". Further, in FIG. 11, the ridge line is formed at the boundary between the lower barrier surface portion 410g and the upper barrier surface portion 420g, but the boundary may be subjected to C chamfering or R chamfering.
[第4實施形態] [Fourth embodiment]
圖12表示第4實施形態之培養容器之要部放大圖。圖12所示之障壁面400h係由下側障壁面部410h及上側障壁面部420h構成。下側障壁面部410h與底面200所成之角度θ 1、和上側障壁面部420h與底面200所成之角度θ 2相同,因此兩面無邊界地連接設置。因此,障壁面400h之構造簡單,就製造上之觀點而言較佳。 Fig. 12 is an enlarged view of a main part of a culture container according to a fourth embodiment. The barrier surface 400h shown in FIG. 12 is composed of a lower barrier surface portion 410h and an upper barrier portion 420h. The angle θ 1 between the lower barrier surface portion 410h and the bottom surface 200 is the same as the angle θ 2 formed by the upper barrier surface portion 420h and the bottom surface 200. Therefore, the two surfaces are connected without borders. Therefore, the structure of the barrier surface 400h is simple, and it is preferable from the viewpoint of manufacturing.
[第5實施形態] [Fifth Embodiment]
圖13表示第5實施形態之培養容器之要部放大圖。圖13所示之障壁面400i之自橫向觀察之剖面形狀為曲面。該曲面之切線與底面200所成之角度θ、θ'、θ"自底面200之側朝向周側壁面300之側逐漸變大(θ<θ'<θ")。藉此,可更確實地使培養細胞不存在於“上下方向觀察時難以觀察到之內部周緣B2之外側部分”,並且障壁面400i之構造本身亦簡單,就製造上之觀點而言較佳。再者,可使角度θ、θ'、θ"於90度以上180度以下之範圍變化。 Fig. 13 is an enlarged view of a main part of a culture container according to a fifth embodiment. The cross-sectional shape of the barrier surface 400i shown in Fig. 13 from the lateral direction is a curved surface. The angles θ, θ', and θ" formed by the tangent of the curved surface and the bottom surface 200 gradually increase from the side of the bottom surface 200 toward the side of the peripheral side wall surface 300 (θ < θ' < θ"). Thereby, it is possible to more reliably prevent the cultured cells from being present in the "outer side portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up-and-down direction", and the structure of the barrier rib surface 400i itself is also simple, and it is preferable from the viewpoint of manufacture. Further, the angles θ, θ', and θ" may be changed in a range of 90 degrees or more and 180 degrees or less.
[第6實施形態] [Sixth embodiment]
圖14表示第6實施形態之培養容器之要部放大圖。圖14所示之障壁面400j係由下側障壁面部410j及上側障壁面部420j構成。下側障壁面部410j之自橫向觀察之剖面形狀為直線,以與底面200呈角度θ 1之方式將內部周緣B2作為基礎端上升,且於高度h1之位置與該剖面形狀為曲線之上側障壁面部420j連接設置。於高度h1之位置,下側障壁面部410j與上側障壁面部420j之邊界形成脊線。 Fig. 14 is an enlarged view of a main part of the culture container according to the sixth embodiment. The barrier surface 400j shown in Fig. 14 is composed of a lower barrier surface portion 410j and an upper barrier portion 420j. The cross-sectional shape of the lower barrier surface portion 410j viewed from the lateral direction is a straight line, and the inner peripheral edge B2 is raised as a base end so as to be at an angle θ1 from the bottom surface 200, and the position at the height h1 and the cross-sectional shape are curved upper side barrier faces. 420j connection settings. At a position of the height h1, a ridge line is formed at a boundary between the lower barrier rib portion 410j and the upper barrier façade 420j.
於本實施形態中,角度θ 1亦可為90度以上且150度以下。藉由為該下限值以上,而自上下方向觀察時,可視野良好地觀察應培養及觀察之內部周緣B2內之整個區域,且藉由為該上限值以下,而可更確實地使培養細胞不存在於“上下方向觀察時難以觀察到之內部周緣B2之外側部分”。上側障壁面部420j之剖面形狀亦可為與第5實施形態中之障壁面400i相同之曲面。 In the present embodiment, the angle θ 1 may be 90 degrees or more and 150 degrees or less. When viewed from the upper and lower sides, the entire area in the inner peripheral edge B2 to be cultured and observed can be observed with good visibility, and can be more reliably made lower than the upper limit value. The cultured cells are not present in the "outer side portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up-and-down direction". The cross-sectional shape of the upper barrier surface portion 420j may be the same curved surface as the barrier surface 400i of the fifth embodiment.
[第7實施形態] [Seventh embodiment]
圖15表示第7實施形態之培養容器之要部放大圖。圖15所示之障壁面400k係由下側障壁面部410k及上側障壁面部420k構成。下側障壁面部410k係以與底面200呈角度θ 1之方式將內部周緣B2作為基礎端而上升,且於高度h1之位置與上側障壁面部420k連接設置。於高度h1之位置,下側障壁面部410k與上側障壁面部420k之邊界形成脊線。上側障壁面部420k相對於底面200以角度θ 2向周側壁面300傾斜,且與周側壁面300形成邊界B3。 Fig. 15 is an enlarged view of a main part of a culture container according to a seventh embodiment. The barrier surface 400k shown in Fig. 15 is composed of a lower barrier surface portion 410k and an upper barrier surface portion 420k. The lower barrier surface portion 410k is raised with the inner peripheral edge B2 as a base end so as to be at an angle θ1 from the bottom surface 200, and is connected to the upper barrier façade 420k at a position of the height h1. At the position of the height h1, the ridge line is formed at the boundary between the lower barrier surface portion 410k and the upper barrier surface portion 420k. The upper barrier surface portion 420k is inclined toward the peripheral side wall surface 300 at an angle θ 2 with respect to the bottom surface 200, and forms a boundary B3 with the circumferential side wall surface 300.
於本實施形態中,角度θ 1亦可為90度以上且150度以下。藉由為該下限值以上,而自上下方向觀察時,可視野良好地觀察應培養及 觀察之內部周緣B2內之整個區域,藉由為該上限值以下,而可減少培養細胞進入“上下方向觀察時難以觀察到之內部周緣B2之外側部分”。 In the present embodiment, the angle θ 1 may be 90 degrees or more and 150 degrees or less. When viewed from the upper and lower directions by being at least the lower limit value, the visual field can be observed well and the culture should be observed. The entire region in the inner peripheral edge B2 of the observation is such that the cultured cells are reduced to the "outer portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up-and-down direction" by being equal to or less than the upper limit value.
角度θ 2並無特別限定。例如存在如下傾向:角度θ 2越大,越可充分確保下側障壁商部410k之高度h1,而減少培養細胞進入“上下方向觀察時難以觀察到之內部周緣B2之外側部分”;角度θ 2越小,越可充分獲取應收容培養液之容積;業者可基於該等兩種傾向而適當決定。 The angle θ 2 is not particularly limited. For example, there is a tendency that the larger the angle θ 2 is, the more the height h1 of the lower barrier quotient portion 410k can be sufficiently ensured, and the cultured cells are reduced to the "outer side portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up and down direction"; the angle θ 2 The smaller the size, the more the volume of the culture solution to be contained can be sufficiently obtained; the operator can appropriately determine the two tendencies based on these two tendencies.
再者,圖15中,下側障壁面部410k與上側障壁面部420k之邊界形成脊線,但亦可對該邊界進行C倒角加工或R倒角加工。 In addition, in FIG. 15, the ridge line is formed in the boundary between the lower side barrier surface portion 410k and the upper side barrier surface portion 420k, but the boundary may be subjected to C chamfering or R chamfering.
<變形例> <Modification>
再者,於上述第2至第7實施形態中,均與第1實施形態同樣地可允許有變形例。即,只要彎液面M(參照圖3至圖9)之至少一部分設置在較內部周緣B2靠外側即可,因此只要邊界E(參照圖3至圖9)未形成於障壁面400f、400g、400h、400i、400j、400k中之下側障壁面410f、410j上,該下側障壁面410f、410j為將內部周緣B2作為基礎端並相對於底面200呈90度上升者,則可允許為任意構造。即,於變形例中,只要以如下方式構成即可:於已收容培養液至培養室C(參照圖3至圖9)內時,培養液之液面S(參照圖3至圖9)與培養容器之內壁之邊界E成為較內部周緣B2更外側。 Further, in the second to seventh embodiments described above, modifications are possible similarly to the first embodiment. In other words, as long as at least a part of the meniscus M (see FIGS. 3 to 9) is provided outside the inner periphery B2, the boundary E (see FIGS. 3 to 9) is not formed on the barrier faces 400f and 400g. 400h, 400i, 400j, 400k, the lower side barrier surfaces 410f, 410j, which are the base end B2 as the base end and rise at 90 degrees with respect to the bottom surface 200, may be allowed to be arbitrary structure. In other words, in the modification, the liquid surface S (see FIGS. 3 to 9) of the culture solution may be configured when the culture solution is stored in the culture chamber C (see FIGS. 3 to 9). The boundary E of the inner wall of the culture vessel becomes outside the inner peripheral edge B2.
[第8實施形態] [Eighth Embodiment]
圖16係第8實施形態之培養容器之示意性外觀立體圖。培養容器1001具有多個培養室C。於本實施形態中,1個培養容器1001具有6個培養室C,但培養室C之數量並不限定於此。例如存在如下傾向:培養室C之數量越 多,越可較佳地享有將已培養之細胞之大部分集中於可觀察之部分的效果;藉由某種程度地限定培養室C之數量,而可適當地獲取用於培養及觀察之底面200上之空間,業者可基於上述情形而適當決定培養室C之數量。作為培養容器1001之培養室C之構造,可應用第1實施形態至第7實施形態以及其等之變形例之任一者中之構造。 Fig. 16 is a schematic perspective view showing the culture container of the eighth embodiment. The culture container 1001 has a plurality of culture chambers C. In the present embodiment, one culture container 1001 has six culture chambers C, but the number of culture chambers C is not limited thereto. For example, there is a tendency that the number of culture chambers C is higher. In many cases, it is preferable to enjoy the effect of concentrating a large part of the cultured cells on the observable portion; by limiting the amount of the culture chamber C to some extent, the bottom surface for cultivation and observation can be appropriately obtained. In the space above 200, the operator can appropriately determine the number of the culture chamber C based on the above situation. The structure of the culture chamber C of the culture container 1001 can be applied to any of the first to seventh embodiments and the modifications thereof.
[實施例] [Examples]
以下示出實施例來具體地說明本發明,但本發明並不限制於下述實施例。 The invention is specifically illustrated by the following examples, but the invention is not limited to the following examples.
<實施例1> <Example 1>
藉由將聚苯乙烯之圓板貼合於使用聚碳酸酯之切削加工品(對無蓋無底圓筒之內周面側進行切削加工而成者),而製作具有圖9所記載之構造之培養容器之試作品(再者,培養容器之正式製品亦可藉由一體成形而製作。以下,將藉由貼合而製作成之試作品亦簡稱為「培養容器」)。θ 1為90度,θ 2為120度。 The structure shown in FIG. 9 was produced by laminating a circular plate of polystyrene to a cut product using polycarbonate (cutting on the inner peripheral side of the bottomless cylinder without a cover). The test piece of the culture container is further produced (in addition, the official product of the culture container can also be produced by integral molding. Hereinafter, the test piece produced by lamination is also simply referred to as "culture container"). θ 1 is 90 degrees and θ 2 is 120 degrees.
於製成之培養容器之培養室添加含10%血清之培養基3mL,自上下方向觀察培養室。圖17表示上下方向觀察照片。如圖17所示,培養容器具有障壁面400e,因此彎液面M之大部分於內部周緣B2a之外側產生,由此可確認於用於培養及觀察之內部周緣B2a之內部可良好地視認之範圍較下述比較例1擴大。 3 mL of a medium containing 10% serum was added to the culture chamber of the prepared culture vessel, and the culture chamber was observed from the up and down direction. Fig. 17 shows a photograph viewed in the up and down direction. As shown in Fig. 17, the culture container has the barrier surface 400e, and therefore most of the meniscus M is generated on the outer side of the inner periphery B2a, whereby it can be confirmed that the inside of the inner periphery B2a for cultivation and observation is well recognized. The range was expanded as compared with Comparative Example 1 below.
於製成之培養容器之培養室添加含10%血清之培養基3mL及人類纖維母細胞38.4萬個,之後於37℃、5%CO2之條件下培養24小時,利用倍率40倍之顯微鏡觀察培養室中央部及周邊部。圖18表示培養室中央 部之顯微鏡觀察照片,圖19表示培養室周邊部之顯微鏡觀察照片。如圖18所示,可確認細胞正常接著於底面200。又,如圖19所示,可確認因障壁面400e而彎液面M之大部分於內部周緣B2a之外側產生,於用於培養及觀察之內部周緣B2a之內部可觀察之範圍較下述比較例1擴大。 3 mL of medium containing 10% serum and 384,000 human fibroblasts were added to the culture chamber of the prepared culture vessel, and then cultured at 37 ° C under 5% CO 2 for 24 hours, and observed under a microscope with a magnification of 40 times. The central part of the room and the surrounding area. Fig. 18 is a microscopic observation photograph of the central portion of the culture chamber, and Fig. 19 is a microscopic observation photograph of the peripheral portion of the culture chamber. As shown in Fig. 18, it was confirmed that the cells were normally followed by the bottom surface 200. Further, as shown in Fig. 19, it can be confirmed that most of the meniscus M is generated on the outer side of the inner peripheral edge B2a due to the barrier surface 400e, and the inner observable range for the inner periphery B2a for culture and observation is compared with the following. Example 1 is expanded.
<實施例2> <Example 2>
藉由將聚苯乙烯之圓板貼合於使用聚碳酸酯之切削加工品(對無蓋無底圓筒之內周面側進行切削加工而成者),而製作具有圖9所記載之構造之培養容器之試作品。本實施例之培養容器除θ 2為150度以外,與實施例1之培養容器相同。 The structure shown in FIG. 9 was produced by laminating a circular plate of polystyrene to a cut product using polycarbonate (cutting on the inner peripheral side of the bottomless cylinder without a cover). Cultivate the test piece of the container. The culture container of this example was the same as the culture container of Example 1 except that θ 2 was 150 degrees.
於製成之培養容器之培養室添加含10%血清之培養基3mL,自上下方向觀察培養室。圖20表示上下方向觀察之照片。如圖20所示,可確認培養容器因具有障壁面400e,而因彎液面M形成之陰影於內部周緣B2a之外側產生,於用於培養及觀察之內部周緣B2a之內部可良好地視認之範圍較下述比較例1擴大。 3 mL of a medium containing 10% serum was added to the culture chamber of the prepared culture vessel, and the culture chamber was observed from the up and down direction. Fig. 20 shows a photograph viewed in the up and down direction. As shown in Fig. 20, it was confirmed that the culture container has the barrier surface 400e, and the shadow formed by the meniscus M is generated on the outer side of the inner periphery B2a, and can be clearly recognized inside the inner periphery B2a for cultivation and observation. The range was expanded as compared with Comparative Example 1 below.
於製成之培養容器之培養室添加含10%血清之培養基3mL及人類纖維母細胞38.4萬個,之後於37℃、5%CO2之條件下培養24小時,利用倍率40倍之顯微鏡觀察培養室中央部及周邊部。圖21表示培養室中央部之顯微鏡觀察照片,圖22表示培養室周邊部之顯微鏡觀察照片。如圖21所示,可確認細胞正常接著於底面200。又,如圖22所示,可確認因障壁面400e而彎液面M之大部分於內部周緣B2a之外側產生,於用於培養及觀察之內部周緣B2a之內部可觀察之範圍較下述比較例1擴大。 3 mL of medium containing 10% serum and 384,000 human fibroblasts were added to the culture chamber of the prepared culture vessel, and then cultured at 37 ° C under 5% CO 2 for 24 hours, and observed under a microscope with a magnification of 40 times. The central part of the room and the surrounding area. Fig. 21 is a photomicrograph of the central portion of the culture chamber, and Fig. 22 is a photomicrograph of the peripheral portion of the culture chamber. As shown in Fig. 21, it was confirmed that the cells were normally followed by the bottom surface 200. Further, as shown in Fig. 22, it can be confirmed that most of the meniscus M is generated on the outer side of the inner peripheral edge B2a due to the barrier surface 400e, and the range which can be observed inside the inner periphery B2a for cultivation and observation is compared with the following. Example 1 is expanded.
若將實施例1與本實施例2之培養室周邊部之顯微鏡觀察照 片(圖19及圖22)進行比較,則可確認實施例2(圖22)中可良好地視認細胞之範圍更廣。於本實施例2中,培養液面S與培養容器100e之內壁面之邊界Ee位於上側周側壁面部420e上,進而,障壁面400e之特定高度hd低於收容於培養室C內之培養液之水位H且亦低於邊界E之高度。因此,與θ 2為120度之實施例1相比,θ 2為150度之本實施例2中於內部周緣B2a之外側產生之彎液面M之範圍變得更大,本實施例2(圖22)中可良好地視認細胞之範圍變得更廣。 Microscopic observation of the peripheral portion of the culture chamber of Example 1 and Example 2 Comparing the sheets (Figs. 19 and 22), it was confirmed that the range of cells that can be clearly visualized in Example 2 (Fig. 22) was wider. In the second embodiment, the boundary Ee between the culture liquid surface S and the inner wall surface of the culture container 100e is located on the upper peripheral side wall surface portion 420e, and further, the specific height hd of the barrier surface 400e is lower than the culture liquid contained in the culture chamber C. The water level H is also below the height of the boundary E. Therefore, the range of the meniscus M generated on the outer side of the inner periphery B2a in the second embodiment is larger as compared with the first embodiment in which θ 2 is 120 degrees, and the present embodiment 2 ( In Fig. 22), the range in which cells can be clearly recognized is made wider.
<比較例1> <Comparative Example 1>
藉由將聚苯乙烯之圓板貼合於聚碳酸酯之無蓋無底圓筒,而製作不具有障壁面之培養容器。將該培養容器之示意性剖面圖示於圖23。 A culture vessel having no barrier surface was produced by laminating a circular plate of polystyrene to a lidless bottomless cylinder of polycarbonate. A schematic cross-sectional view of the culture vessel is shown in FIG.
於製成之培養容器之培養室添加含10%血清之培養基3mL,自上下方向觀察培養室。圖24表示上下方向觀察之照片。如圖24所示,彎液面M全部於周緣B之內側產生,於用於培養及觀察之周緣B之內部確認到因彎液面M形成之陰影。 3 mL of a medium containing 10% serum was added to the culture chamber of the prepared culture vessel, and the culture chamber was observed from the up and down direction. Fig. 24 shows a photograph viewed in the up and down direction. As shown in Fig. 24, the meniscus M was all generated inside the peripheral edge B, and the shadow formed by the meniscus M was confirmed inside the periphery B for cultivation and observation.
於製成之培養容器之培養室添加含10%血清之培養基3mL及人類纖維母細胞38.4萬個,之後於37℃、5%CO2之條件下培養24小時,利用倍率40倍之顯微鏡觀察培養室中央部及周邊部。圖25表示培養室中央部之顯微鏡觀察照片,圖26表示培養室周邊部之顯微鏡觀察照片。如圖25所示,可確認細胞正常接著於底面200。又,如圖26所示,可確認彎液面M全部於周緣B之內側產生,於用於培養及觀察之周緣B之內部產生因彎液面M形成之陰影,難以觀察周緣B附近之細胞,於用於培養及觀察之周緣B之內部可觀察之範圍較窄。 3 mL of medium containing 10% serum and 384,000 human fibroblasts were added to the culture chamber of the prepared culture vessel, and then cultured at 37 ° C under 5% CO 2 for 24 hours, and observed under a microscope with a magnification of 40 times. The central part of the room and the surrounding area. Fig. 25 is a photomicrograph of the central portion of the culture chamber, and Fig. 26 is a photomicrograph of the peripheral portion of the culture chamber. As shown in Fig. 25, it was confirmed that the cells were normally followed by the bottom surface 200. Further, as shown in Fig. 26, it was confirmed that all of the meniscus M was generated inside the peripheral edge B, and a shadow formed by the meniscus M was generated inside the periphery B for culture and observation, and it was difficult to observe cells near the periphery B. The range that can be observed inside the periphery B used for culture and observation is narrow.
[第1實施形態至第8實施形態中之各部與請求項之各構成要素之對應關係] [Correspondence between each component in the first embodiment to the eighth embodiment and each component of the request item]
於本發明中,培養容器100、1001相當於「培養容器」,底面200相當於「底面」,周側壁面300、300b、300d相當於「周側壁面」,障壁面400、400a、400b、400c、400d、400e、400f、400g、400h、400i、400j、400k相當於「障壁面」,下側障壁面部410、410f、410g、410h、410j、410k相當於「下側障壁面部」,上側障壁面部420、420d、420e、420f、420g、420h、420j、420k相當於「上側障壁面部」,周緣O相當於「周緣」,內部周緣B2、B2a相當於「內部周緣」,邊界B3、B3b、B3d相當於(障壁面與周側壁面之)「邊界」,培養室C相當於「培養室」,角度θ 1相當於(下側障壁面部與底面所成之)「角度」,角度θ、θ'、θ"相當於(障壁面之剖面形狀之切線與底面所成之)「角度」。 In the present invention, the culture containers 100 and 1001 correspond to a "culture container", the bottom surface 200 corresponds to the "bottom surface", and the peripheral side wall surfaces 300, 300b, and 300d correspond to the "peripheral side wall surface", and the barrier surfaces 400, 400a, 400b, and 400c 400d, 400e, 400f, 400g, 400h, 400i, 400j, and 400k correspond to the "barrier surface", and the lower barrier surface portions 410, 410f, 410g, 410h, 410j, and 410k correspond to the "lower barrier surface portion" and the upper barrier surface portion. 420, 420d, 420e, 420f, 420g, 420h, 420j, and 420k correspond to "upper barrier face", peripheral edge O corresponds to "perimeter", inner periphery B2, B2a corresponds to "internal circumference", and boundaries B3, B3b, and B3d are equivalent. In the "boundary" of the (barrier surface and the peripheral side wall surface), the culture chamber C corresponds to the "culture chamber", and the angle θ 1 corresponds to the "angle" (the angle formed by the lower barrier surface portion and the bottom surface), the angle θ, θ', θ" corresponds to (the angle formed by the tangent to the bottom surface of the cross-sectional shape of the barrier surface).
本發明之較佳之實施形態如上所述,但本發明並不僅限定於該等實施形態,存在其他不脫離本發明之主旨及範圍之各種實施形態。進而,於本實施形態中敍述之作用及效果為一例,並非限定本發明。 The preferred embodiments of the present invention are as described above, but the present invention is not limited to the embodiments, and various other embodiments are possible without departing from the spirit and scope of the invention. Furthermore, the actions and effects described in the present embodiment are merely examples and are not intended to limit the present invention.
[產業上之可利用性] [Industrial availability]
根據本發明,可提供一種即便產生彎液面,因該彎液面而形成之陰影亦不會妨礙培養細胞之觀察之培養容器。 According to the present invention, it is possible to provide a culture container in which the shadow formed by the meniscus does not interfere with the observation of the cultured cells even if a meniscus is generated.
100‧‧‧培養容器 100‧‧‧ culture container
200‧‧‧底面 200‧‧‧ bottom
300‧‧‧周側壁面 300‧‧‧ Weekly side wall
400‧‧‧障壁面 400‧‧ ‧ barrier wall
B2‧‧‧(底面之)內部周緣 B2‧‧‧ (bottom) internal circumference
B3‧‧‧(障壁面與周側壁面之)邊界 B3‧‧‧ (Bound wall and circumferential side wall) boundary
C‧‧‧培養室 C‧‧‧Cultivation room
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US4321330A (en) * | 1980-04-04 | 1982-03-23 | Baker Fraser L | Tissue culture device |
WO1983000047A1 (en) * | 1981-06-22 | 1983-01-06 | American Micro Scan Inc | Improved biomedical analysis tray |
JPS6251977A (en) * | 1985-09-02 | 1987-03-06 | Terumo Corp | Tissue culture plate |
WO1991006624A1 (en) * | 1989-10-26 | 1991-05-16 | Costar Corporation | Dish for in vitro fertilization |
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