A culture device, an exosome culture, separation and collection system and method

Abstract

This invention relates to a culture device, an exosome culture separation and collection system, and a method. The culture device includes a box unit, a support unit, a culture unit, a first vibration unit, a first delivery tube unit, a second delivery tube unit, a third delivery tube unit, and a temperature control unit. Its advantages lie in that the first vibration element provides sustained vibration, causing the cell culture medium in the culture unit to flow vertically, while the second vibration element provides intermittent vibration, causing the cell culture medium in the culture unit to flow horizontally in a ripple pattern, thus improving culture efficiency and shortening culture time.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cell separation, and in particular to a culture device, an exosome culture separation and collection system and method. BACKGROUND

[0002] Exosomes are extracellular vesicles with a lipid bilayer, about 30-150nm in diameter, which can be applied to immunotherapy and tumor therapy, and have good development prospects in biomedicine.

[0003] Although exosomes have good application effects, they cannot be cultured and separated on a large scale. At present, only small equipment can be used for trace culture and filtration of exosomes, resulting in high costs.

[0004] The existing cell culture device is generally divided into a stirring type cell culture device and a non-stirring type cell culture device. For the stirring type cell culture device, although the device is simple to operate and has low cost, the exosomes are easily damaged. For the non-stirring type cell culture device, although the exosomes can be avoided from being damaged, the culture efficiency is low, the device is complex, and the cost is high.

[0005] In addition, the exosome separation equipment can only be used for trace separation, and cannot meet the large-scale separation. This results in low separation efficiency, long separation time, and high use cost.

[0006] At present, there is no effective solution to the problems of exosomes not being easy to culture, being difficult to separate in large quantities, and the entire process being costly in the related art. SUMMARY

[0007] The present application aims to solve the problems of exosomes not being easy to culture, being difficult to separate in large quantities, and the entire process being costly in the related art by providing a culture device, an exosome culture separation and collection system and method.

[0008] To achieve the above-mentioned purpose, the technical solution adopted by the present application is as follows:

[0009] In a first aspect, a culture device for an exosome culture separation and collection system is provided, comprising:

[0010] a box unit placed on a horizontal plane;

[0011] a support unit arranged inside the box unit;

[0012] a culture unit detachably arranged inside the support unit;

[0013] A first vibration unit is disposed on the support unit and connected to the culture unit, and is used to vibrate the culture unit.

[0014] A temperature control unit, located inside the enclosure unit, is used to regulate the internal temperature of the enclosure unit;

[0015] The first vibration unit includes:

[0016] A plurality of first vibration elements are arranged at intervals along the vertical direction of the support unit inside the support unit for vibrating the bottom of the culture unit;

[0017] A plurality of first rotating elements are distributed at the bottom of the first vibrating element;

[0018] A plurality of second vibration elements are distributed at the bottom of the first vibration element and are used to vibrate the interior of the culture unit;

[0019] A plurality of second rotating elements are distributed at the ends of the second vibrating element and connected to the corresponding first rotating elements.

[0020] In some embodiments, the box unit includes:

[0021] A housing element, the housing element being placed on a horizontal plane;

[0022] A buffer element is disposed at the bottom of the housing element to reduce the impact caused by external vibrations.

[0023] In some embodiments, the support unit includes:

[0024] A support element, wherein the support element is disposed inside the housing unit;

[0025] A plurality of shelf elements are arranged at intervals inside the support element along the vertical direction of the support element, and the shelf elements are provided with the first vibration unit;

[0026] A plurality of first limiting elements are symmetrically arranged on the inner sidewalls of the left and right sides of the support element for detachable connection with the culture unit.

[0027] A plurality of first locking elements are distributed on the front and rear sides of the corresponding shelf element for detachable connection with the culture unit;

[0028] A plurality of first positioning elements are connected to corresponding first locking elements for detachable connection with the culture unit.

[0029] In some embodiments, the culture unit includes:

[0030] A plurality of culture elements are arranged at intervals inside the support unit along the vertical direction of the support unit and are detachably connected to the support unit;

[0031] A plurality of second limiting elements are distributed on the left and right sides of the corresponding culture element and are detachably connected to the support unit;

[0032] A plurality of second locking elements are distributed on the front and rear sides of the corresponding culture element and are detachably connected to the support unit;

[0033] A plurality of second positioning elements are distributed on the front and rear sides of the corresponding culture element and are detachably connected to the second locking element.

[0034] A plurality of third locking elements, wherein the plurality of third locking elements are detachably connected to the corresponding second locking element and the corresponding second positioning element, and are detachably connected to the corresponding second locking element and the bracket unit.

[0035] In some embodiments, the temperature control unit includes:

[0036] A temperature monitoring element is disposed inside the enclosure unit and is used to monitor the internal temperature of the enclosure unit;

[0037] A temperature regulating element is disposed inside the box unit and is used to regulate the internal temperature of the box unit.

[0038] Secondly, an exosome culture, isolation, and collection system is provided, comprising:

[0039] The culture device as described in the first aspect is provided with a cell culture medium inside for culturing exosomes;

[0040] A separation device, connected to the culture device, is used to perform multi-stage separation of the cell culture medium delivered by the culture device to obtain exosomes and secondary cell culture medium;

[0041] A collection device, connected to the separation device, is used to collect the exosomes separated by the separation device;

[0042] A rinsing device is connected to the culture device and the separation device respectively, and is used to provide rinsing solution to the culture device and the separation device respectively, for rinsing the cell culture medium of the culture device to the separation device and for separation by the separation device, wherein the composition of the rinsing solution is the same as that of the cell culture medium;

[0043] A reflux device is connected to the culture device and the separation device respectively, and is used to transport the secondary cell culture medium of the separation device to the culture device;

[0044] A control device is connected to the culture device, the separation device, the collection device, the rinsing device, and the reflux device, respectively.

[0045] In some embodiments, the separation device includes:

[0046] A separation unit is connected to the culture device, the collection device, the rinsing device, and the reflux device, respectively.

[0047] A vacuum unit is disposed on both sides of the separation unit and connected to the control device and the separation unit respectively, and is used to provide negative pressure to the separation unit under the control of the control device;

[0048] An electrode unit is disposed on both sides of the separation unit and connected to the control device and the separation unit respectively, and is used to alternately generate an electric field inside the separation unit under the control of the control device;

[0049] The second vibration unit is connected to the separation unit and the control device respectively, and is used to vibrate the separation unit.

[0050] Wherein, while the vacuum unit provides negative pressure to one side of the separation unit, the vacuum unit does not provide negative pressure to the other side of the separation unit.

[0051] In some embodiments, the separation unit includes:

[0052] A first chamber element is connected to the culture device, the rinsing device, and the reflux device, respectively.

[0053] A second chamber element is symmetrically disposed on both sides of the first chamber element and communicates with the first chamber element;

[0054] The third chamber element is symmetrically disposed on both sides of the second chamber element, communicates with the second chamber element, and is also communicated with the vacuum unit.

[0055] A first filter element is disposed between the first chamber element and the second chamber element;

[0056] The second filter element is disposed between the second chamber element and the third chamber element, and the pore size of the second filter element is smaller than that of the first filter element.

[0057] In some embodiments, the vacuum unit includes:

[0058] A first vacuum element is disposed on one side of the separation unit and is connected to both the separation unit and the control device.

[0059] A second vacuum element is disposed on the other side of the separation unit and is connected to the separation unit and the control device, respectively.

[0060] The first vacuum element and the second vacuum element alternately provide negative pressure to the separation unit.

[0061] In some embodiments, the electrode unit includes:

[0062] The first electrode element is disposed on one side inside the separation unit and is connected to the separation unit and the control device respectively;

[0063] The second electrode element is disposed on the other side inside the separation unit and is connected to the separation unit and the control device respectively;

[0064] The first electrode element and the second electrode element alternately generate an electric field inside the separation unit.

[0065] In some embodiments, the second vibration unit includes:

[0066] A third vibration element is disposed on the side of the separation unit and connected to the control device;

[0067] A fourth vibration element is disposed on the side of the separation unit and connected to the control device.

[0068] In some embodiments, the collection device includes:

[0069] A first collection unit is connected to the separation device and the control device respectively, and is used to collect exosomes obtained by the separation device in the first stage of separation.

[0070] The second collection unit is connected to the separation device and the control device, respectively, and is used to collect exosomes obtained by the separation device through multi-stage separation.

[0071] In some embodiments, the first collection unit includes:

[0072] A first liquid delivery element is connected to the separation device and the control device respectively, and is used to deliver the exosomes obtained by the primary separation of the separation device.

[0073] At least one first storage element, the first storage element being connected to the first liquid delivery element, is used to collect exosomes obtained by primary separation in the separation device;

[0074] At least one first liquid level monitoring element is disposed inside the first storage element and connected to the control device.

[0075] In some embodiments, the second collection unit includes:

[0076] A second liquid delivery element is connected to the separation device and the control device respectively, and is used to deliver exosomes obtained by multi-stage separation by the separation device.

[0077] At least one second storage element, the second storage element being connected to the second liquid delivery element, for collecting exosomes obtained through multi-stage separation by the separation device;

[0078] At least one second liquid level monitoring element is disposed inside the second storage element and connected to the control device.

[0079] In some embodiments, the rinsing device includes:

[0080] The first liquid storage unit is disposed on one side of the culture device and is used to store the rinsing solution;

[0081] The first rinsing unit is connected to the first liquid storage unit, the culture device, and the control device, respectively, and is used to provide rinsing solution to the culture device to rinse the cell culture medium of the culture device to the separation device;

[0082] The second rinsing unit is connected to the first liquid storage unit, the separation device, and the control device, respectively, and is used to provide rinsing liquid to the separation device for separation.

[0083] In some embodiments, the first liquid storage unit includes:

[0084] A third storage element is disposed on one side of the culture device and is used to store rinsing solution;

[0085] The third liquid level monitoring element is disposed inside the third storage element and connected to the control device.

[0086] In some embodiments, the first rinsing unit includes:

[0087] A third liquid delivery element is connected to the first liquid storage unit, the culture device, and the control device, respectively.

[0088] A first flow monitoring element is disposed on the third liquid delivery element and connected to the control device.

[0089] In some embodiments, the second rinsing unit includes:

[0090] A fourth liquid delivery element is connected to the first liquid storage unit, the separation device, and the control device, respectively.

[0091] A second flow monitoring element is disposed on the fourth liquid delivery element and connected to the control device.

[0092] In some embodiments, the recirculation device includes:

[0093] A filtration unit, which is connected to the separation device;

[0094] The second liquid storage unit is disposed on one side of the filter unit and connected to the filter unit;

[0095] The third liquid storage unit is disposed on one side of the filter unit and connected to the filter unit;

[0096] A reflux unit is connected to the culture device, the second liquid storage unit, and the control device, respectively, and is used to reflux the secondary cell culture medium obtained by the separation device back to the culture device.

[0097] In some embodiments, the filtering unit includes:

[0098] The fifth liquid delivery element is connected to both the separation device and the control device.

[0099] The fourth storage element is connected to the fifth liquid delivery element, the second liquid storage unit, and the third liquid storage unit respectively;

[0100] An electrostatic adsorption element is disposed inside the fourth storage element and connected to the control device.

[0101] The third filter element is disposed inside the fourth storage element and is located downstream of the electrostatic adsorption element.

[0102] In some embodiments, the second liquid storage unit includes:

[0103] The sixth liquid delivery element is connected to the filter unit and the control device respectively;

[0104] The fifth storage element is connected to the sixth liquid delivery element and the reflux unit.

[0105] In some embodiments, the third liquid storage unit includes:

[0106] A seventh liquid delivery element, which is connected to the filter unit and the control device respectively;

[0107] A sixth storage element, which is connected to the seventh liquid delivery element.

[0108] In some embodiments, the reflow unit includes:

[0109] The eighth liquid delivery element is connected to the second liquid storage unit, the culture device, and the control device respectively;

[0110] A third flow monitoring element is disposed on the eighth liquid delivery element and connected to the control device.

[0111] Thirdly, a method for culturing, isolating, and collecting exosomes is provided, applied to the exosome culture, isolating, and collecting system as described in the second aspect, comprising:

[0112] Cell culture medium is placed into the culture device for culturing exosomes;

[0113] When the culture time of the culture device reaches a preset culture time threshold, the control device controls the rinsing device to provide rinsing fluid to the interior of the culture device so that all the cell culture medium of the culture device enters the separation device, wherein the composition of the rinsing fluid is the same as the composition of the cell culture medium;

[0114] The control device controls the separation device to perform multi-stage separation of the cell culture medium to obtain exosomes and secondary cell culture medium. In the case of multi-stage separation, the control device controls the rinsing device to provide rinsing fluid to the interior of the separation device.

[0115] After the separation device has completed multi-stage separation, the control device controls the collection device to collect the exosomes obtained by the separation device.

[0116] After the separation device completes the primary separation, the control device controls the reflux device to return the secondary cell culture medium obtained from the primary separation to the culture device for use by the culture device to culture exosomes.

[0117] In some of these embodiments, the following are included:

[0118] Cell culture medium is placed into the culture unit of the culture device for culturing exosomes;

[0119] The control device controls the first vibration unit of the culture device to vibrate the culture unit in order to increase the culture rate;

[0120] When the culture time of the culture unit reaches a preset culture time threshold, the control device controls the first rinsing unit of the rinsing device to provide rinsing fluid to the interior of the culture unit, so that the cell culture medium of the culture unit enters the separation unit of the separation device;

[0121] The control device controls the vacuum unit, electrode unit, and second vibration unit of the separation device to perform multi-stage separation of the cell culture medium in the separation unit to obtain exosomes and secondary cell culture medium. In the case of multi-stage separation in the separation unit, the control device controls the second rinsing unit of the rinsing device to provide rinsing solution to the separation unit.

[0122] After the separation unit has completed multi-stage separation, the control device controls the first collection unit of the collection device to collect the exosomes separated in the first stage, and the second collection unit of the collection device to collect the exosomes separated in the multi-stage separation.

[0123] After the separation unit completes the primary separation, the control device controls the filtration unit of the reflux device to filter the secondary cell culture medium.

[0124] The control device controls the second liquid storage unit of the reflux device to collect the secondary cell culture medium filtered by the filtration unit;

[0125] The control device controls the third storage unit of the reflux device to collect the secondary cell culture medium that has not been filtered by the filtration unit;

[0126] The control device controls the reflux unit of the reflux device to reflux the secondary cell culture medium collected by the second storage unit back to the culture unit.

[0127] The present invention adopts the above technical solution and has the following technical effects compared with the prior art:

[0128] The present invention discloses a culture device, an exosome cell culture separation and collection system and method, which utilizes a first vibration element to vibrate for a long time to make the cell culture medium in the culture unit flow vertically, and utilizes a second vibration element to vibrate intermittently to make the cell culture medium in the culture unit flow horizontally in a ripple pattern, thereby improving culture efficiency and shortening culture time. Attached Figure Description

[0129] Figure 1 This is a schematic diagram of a culture apparatus according to an embodiment of the present invention;

[0130] Figure 2 This is a schematic diagram of a box unit according to an embodiment of the present invention;

[0131] Figure 3 This is a schematic diagram of a support unit according to an embodiment of the present invention;

[0132] Figure 4 This is a schematic diagram of a culture unit according to an embodiment of the present invention;

[0133] Figure 5 This is a schematic diagram of a vibration unit according to an embodiment of the present invention;

[0134] Figure 6 This is a schematic diagram of an exosome culture, separation, and collection system according to an embodiment of the present invention;

[0135] Figure 7 This is a schematic diagram of a separation device according to an embodiment of the present invention;

[0136] Figure 8 This is a schematic diagram of a collection device according to an embodiment of the present invention;

[0137] Figure 9This is a schematic diagram of a rinsing device according to an embodiment of the present invention;

[0138] Figure 10 This is a schematic diagram of a reflux device according to an embodiment of the present invention.

[0139] The reference numerals in the accompanying drawings are as follows: 100, culture device; 110, box unit; 111, box element; 112, buffer element; 120, support unit; 121, support element; 122, shelf element; 123, first limiting element; 124, first locking element; 125, first positioning element; 130, culture unit; 131, culture element; 132, second limiting element; 133, second locking element; 134, second positioning element; 135, third locking element; 140, first vibration unit; 141, first vibration element; 142, first rotating element; 143, second vibration element; 144, second rotating element; 150, first delivery tube unit; 160, second delivery tube unit; 170, third delivery tube unit;

[0140] 200. Separation device; 210. Separation unit; 211. First chamber element; 212. Second chamber element; 213. Third chamber element; 214. First filter element; 215. Second filter element; 220. Vacuum unit; 221. First vacuum element; 222. Second vacuum element; 230. Electrode unit; 231. First electrode element; 232. Second electrode element; 240. Second vibration unit; 241. Third vibration element; 242. Fourth vibration element; 250. Fourth delivery pipe unit; 260. Fifth delivery pipe unit; 270. Sixth delivery pipe unit; 280. Seventh delivery pipe unit;

[0141] 300. Collection device; 310. First collection unit; 311. First liquid conveying element; 312. First storage element; 320. Second collection unit; 321. Second liquid conveying element; 322. Second storage element; 330. Eighth conveying pipe unit; 340. Ninth conveying pipe unit;

[0142] 400. Flushing device; 410. First liquid storage unit; 411. Third storage element; 420. First flushing unit; 421. Third liquid delivery element; 430. Second flushing unit; 431. Fourth liquid delivery element; 440. Tenth delivery pipe unit; 450. Eleventh delivery pipe unit;

[0143] 500. Reflux device; 510. Filter unit; 511. Fifth liquid conveying element; 512. Fourth storage element; 513. Electrostatic adsorption element; 514. Third filter element; 520. Second liquid storage unit; 521. Sixth liquid conveying element; 522. Fifth storage element; 530. Third liquid storage unit; 531. Seventh liquid conveying element; 532. Sixth storage element; 540. Reflux unit; 541. Eighth liquid conveying element; 550. Twelfth conveying pipe unit; 560. Thirteenth conveying pipe unit; 570. Fourteenth conveying pipe unit; 580. Fifteenth conveying pipe unit. Detailed Implementation

[0144] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0145] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.

[0146] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but this is not intended to limit the scope of the invention.

[0147] Example 1

[0148] like Figure 1 As shown, a culture device 100 is used for an exosome culture, separation, and collection system, including a box unit 110, a support unit 120, a culture unit 130, a first vibration unit 140, and a temperature control unit. The box unit 110 is placed on a horizontal plane; the support unit 120 is disposed inside the box unit 110; the culture unit 130 is detachably disposed inside the support unit 120; the first vibration unit 140 is disposed in the support unit 120 and connected to the culture unit 130, for vibrating the culture unit 130; and the temperature control unit is disposed inside the box unit 110 for regulating the internal temperature of the box unit 110.

[0149] like Figure 2As shown, the box unit 110 includes a box element 111, a buffer element 112, a first inlet element, a second inlet element, a first outlet element, a first valve element, a second valve element, and a third valve element. The box element 111 is placed on a horizontal plane; the buffer element 112 is located at the bottom of the box element 111 to reduce the impact of external vibrations; the first inlet element is located on one side of the box element 111; the second inlet element is located on one side of the box element 111; the first outlet element is located on one side of the box element 111; the first valve element is located on the first inlet element; the second valve element is located on the second inlet element; and the third valve element is located on the first outlet element.

[0150] The box element 111 includes a box body and a box door, which can be opened and closed to facilitate the replacement of the culture unit 130 placed inside.

[0151] The buffer element 112 is disposed on the inner and / or outer side of the bottom of the cover box element 111.

[0152] In some of these embodiments, the buffer element 112 is made of materials such as rubber or silicone.

[0153] The first inlet element is located on the top or right side of the box element 111; the second inlet element is located on the top or right side of the box element 111 and is situated on one side of the first inlet element; the first outlet element is located on the left side of the box element 111.

[0154] In some embodiments, the first inlet element is a first liquid inlet, with interfaces for connecting to a liquid delivery pipe on both its inner and outer sides; the second inlet element is a second liquid inlet, with interfaces for connecting to a liquid delivery pipe on both its inner and outer sides; and the first outlet element is a first liquid outlet, with interfaces for connecting to a liquid delivery pipe on both its inner and outer sides.

[0155] The first valve element is located inside the first inlet element and is used to control the opening and closing of the first inlet element; the second valve element is located inside the second inlet element and is used to control the opening and closing of the second inlet element; the third valve element is located inside the first outlet element and is used to control the opening and closing of the first outlet element.

[0156] In some of these embodiments, the first valve element is a first solenoid valve; the second valve element is a second solenoid valve; and the third valve element is a third solenoid valve.

[0157] like Figure 3As shown, the support unit 120 includes a support element 121, a plurality of shelf elements 122, a plurality of first limiting elements 123, a plurality of first locking elements 124, and a plurality of first positioning elements 125. The support element 121 is disposed inside the box unit 110; the plurality of shelf elements 122 are spaced apart along the vertical direction of the support element 121 inside the support element 121, and each shelf element 122 is provided with a first vibration unit 140; the plurality of first limiting elements 123 are symmetrically disposed on the inner sidewalls of the left and right sides of the support element 121 for detachable connection with the culture unit 130; the plurality of first locking elements 124 are distributed on the front and rear sides of the corresponding shelf elements 122 for detachable connection with the culture unit 130; and the plurality of first positioning elements 125 are connected to the corresponding first locking elements 124 for detachable connection with the culture unit 130.

[0158] Specifically, the support element 121 is disposed inside the box element 111; the first limiting element 123 is slidably connected to the culture unit 130.

[0159] The support element 121 has a support, a top plate, a bottom plate, a front side plate and a rear side plate. The top plate, bottom plate, front side plate and rear side plate are respectively set at the top, bottom, front and rear of the support. Only the left and right sides of the support are left empty to facilitate the installation of the culture unit 130.

[0160] Several shelf elements 122 are fixedly disposed inside the support element 121. The shelf element 122 located at the bottom is at a certain distance from the bottom of the support element 121; the shelf element 122 located at the top is at a certain distance from the top of the support element 121.

[0161] At least one first limiting element 123 is symmetrically provided on the inner sidewalls of the support element 121 located between adjacent shelf elements 122. Specifically, if the number of shelf elements 122 is n (n≥2), then the number of first limiting elements 123 is at least 2 (n-1).

[0162] In some embodiments, the first limiting element 123 consists of at least two limiting rods. The first ends of the two limiting rods are connected, and the second ends of the two limiting rods are separated.

[0163] In some embodiments, the first limiting element 123 is triangular, meaning one limiting rod is arranged in parallel, and the remaining two limiting rods are symmetrically arranged on both sides of the limiting rod with the limiting rod as the center. Alternatively, the three limiting rods may be parallel to each other, or the remaining two limiting rods may be inclined outwards.

[0164] In some of these embodiments, the cross-section of the limiting rod is rectangular or L-shaped.

[0165] At least one first locking element 124 is provided on each of the left and right sides of each shelf element 122 to lock the culture unit 130 and prevent the culture unit 130 from falling.

[0166] In some of these embodiments, the first locking element 124 is a locking groove.

[0167] At least one first positioning element 125 is provided in each shelf element 122, and the first positioning element 125 is connected to the first locking elements 124 on the front and rear sides of the shelf element 122.

[0168] In some of these embodiments, the first positioning element 125 is a first positioning hole.

[0169] like Figure 4 As shown, the culture unit 130 includes a plurality of culture elements 131, a plurality of second limiting elements 132, a plurality of second locking elements 133, a plurality of second positioning elements 134, a plurality of third locking elements 135, a plurality of third inlet elements, a plurality of fourth inlet elements, a plurality of second outlet elements, a plurality of fourth valve elements, a plurality of fifth valve elements, and a plurality of sixth valve elements. The culture elements 131 are spaced apart along the vertical direction of the support unit 120 inside the support unit 120 and are detachably connected to the support unit 120; the second limiting elements 132 are distributed on the left and right sides of the corresponding culture elements 131 and are detachably connected to the support unit 120; the second locking elements 133 are distributed on the front and rear sides of the corresponding culture elements 131 and are detachably connected to the support unit 120; the second positioning elements 134 are distributed on the front and rear sides of the corresponding culture elements 131 and are detachably connected to the second locking elements 135; the third limiting elements 131 are spaced apart along the vertical direction of the support unit 120 and are detachably connected to the support unit 120; the second limiting elements 132 are distributed on the front and rear sides of the corresponding culture elements 131 and are detachably connected to the second locking elements 133; the second limiting elements 132 are spaced apart along the vertical direction of the support unit 120 and are detachably connected to the support unit 120; the second limiting elements 132 are distributed on the front and rear sides of the corresponding culture elements 131 and are detachably connected to the second locking elements 135; the second limiting elements 132 are spaced apart along the vertical direction of the support unit 120 and are detachably connected to the support unit 120; the second limiting elements 132 are spaced apart along the vertical direction of the support unit 120 and are detachably connected to the support unit Locking element 135 is detachably connected to the corresponding second locking element 133 and the corresponding second positioning element 134, and is also detachably connected to the corresponding second locking element 133 and the support unit 120; a plurality of third inlet elements are distributed on one side of the corresponding culture element 131; a plurality of fourth inlet elements are distributed on one side of the corresponding culture element 131; a plurality of second outlet elements are distributed on one side of the corresponding culture element 131; a plurality of fourth valve elements are disposed on the corresponding third inlet elements; a plurality of fifth valve elements are disposed on the corresponding fourth inlet elements; and a plurality of sixth valve elements are disposed on the corresponding second outlet elements.

[0170] Specifically, the culture element 131 is disposed on the upper part of the corresponding shelf element 122; the second limiting element 132 is detachably slidably connected to the corresponding first limiting element 123; the second locking element 133 is detachably connected to the corresponding first locking element 124; the second positioning element 134 corresponds to the corresponding first positioning element 125; the third locking element 135 is detachably connected to the corresponding first positioning element 125 and the second positioning element 134 respectively; the third inlet element corresponds to the first inlet element; the fourth inlet element corresponds to the second inlet element; and the second outlet element corresponds to the first outlet element.

[0171] The culture element 131 is a culture frame with an open top, in which cell culture medium is placed for exosome cell culture, and its bottom and interior are vibrated by the first vibration unit 140.

[0172] The height of the culture element 131 is the same as the distance between two adjacent shelf elements 122.

[0173] The difference between the number of culture elements 131 and the number of shelf elements 122 is 1, i.e., n-1.

[0174] At least one second limiting element 132 is symmetrically arranged on the front and rear sides of each culture element 131. The number of second limiting elements 132 is the same as the number of first limiting elements 123, and the shape of the second limiting elements 132 is the same as the shape of the first limiting elements 123.

[0175] In some of these embodiments, the second limiting element 132 is a limiting groove.

[0176] At least one second locking element 133 is symmetrically arranged on the left and right sides of each culture element 131, and the second locking element 133 is slidably connected or rotatably connected to the corresponding culture element 131.

[0177] In some of these embodiments, the second locking element 133 is a locking plate.

[0178] At least one second positioning element 134 is provided in each culture element 131 and each second locking element 133. Specifically, each culture element 131 has a second positioning element 134, and each of the second locking elements 133 on its left and right sides has a second positioning element 134. When the second locking element 133 is not connected to the first locking element 124, the second positioning element 134 of the second locking element 133 is connected to the second positioning element 134 of the culture element 131. When the second locking element 133 is connected to the first locking element 124, the second positioning element 134 of the second locking element 133 is connected to the first positioning element 125 and is not connected to the second positioning element 134 of the culture element 131.

[0179] The second positioning element 134 of the culture element 131 is disposed through the culture element 131.

[0180] In some of these embodiments, the second positioning element 134 is a second positioning hole.

[0181] For each culture element 131, at least one third locking element 135 is provided. When the second locking element 133 is not connected to the first locking element 124, the third locking element 135 is detachably connected to the second positioning element 134 of the culture element 131 and the second positioning element 134 of the second locking element 133; when the second locking element 133 is connected to the first locking element 124, the third locking element 135 is detachably connected to the first positioning element 125 and the second positioning element 134 of the second locking element 133.

[0182] In some embodiments, the third locking element 135 includes a locking lever and a locking nut, the locking nut being detachably disposed at both ends of the locking lever.

[0183] Through the limiting sliding of the first limiting element 123 and the second limiting element 132, and the locking cooperation of the first locking element 124, the second locking element 133 and the third locking element 135, the culture element 131 is firmly fixed inside the support element 121 and is set close to the shelf element 122.

[0184] Each culture element 131 is provided with a third inlet element, a fourth inlet element, and a second outlet element. The third inlet element is located on the left side of the culture element 131, the fourth inlet element is located on the left side of the culture element 131, and the second outlet element is located on the right side of the culture element 131.

[0185] In some embodiments, the third inlet element is a third liquid inlet, and its outer side is provided with an interface for connecting to a liquid delivery pipe; the second outlet element is a second liquid outlet, and its outer side is provided with an interface for connecting to a liquid delivery pipe.

[0186] The fourth valve element is located inside the third inlet element and is used to control the opening and closing of the third inlet element; the fifth valve element is located inside the fourth inlet element and is used to control the opening and closing of the fourth inlet element; the sixth valve element is located inside the second outlet element and is used to control the opening and closing of the second outlet element.

[0187] In some of these embodiments, the fourth valve element is a fourth solenoid valve; the fifth valve element is a fifth solenoid valve; and the sixth valve element is a sixth solenoid valve.

[0188] like Figure 5 As shown, the first vibration unit 140 includes a plurality of first vibration elements 141, a plurality of first rotating elements 142, a plurality of second vibration elements 143, and a plurality of second rotating elements 144. The plurality of first vibration elements 141 are spaced apart along the vertical direction of the support unit 120 inside the support unit 120 to vibrate the bottom of the culture unit 130; the plurality of first rotating elements 142 are distributed at the bottom of the first vibration elements 141; the plurality of second vibration elements 143 are distributed at the bottom of the first vibration elements 141 to vibrate inside the culture unit 130; and the plurality of second rotating elements 144 are distributed at the ends of the second vibration elements 143 and connected to the corresponding first rotating elements 142.

[0189] Specifically, the first vibration element 141 is disposed at the bottom of the corresponding shelf element 122 and is used to vibrate the bottom of the culture element 131; the second vibration element 143 vibrates the interior of the culture element 131.

[0190] The first vibration element 141 is disposed at the bottom of the shelf element 122.

[0191] The number of first vibration elements 141 is the same as the number of shelf elements 122.

[0192] In some of these embodiments, the first vibrating element 141 is a vibrating plate.

[0193] At least five first rotating elements 142 are distributed at the bottom of each first vibrating element 141, wherein the first vibrating element 141 at the bottommost position does not have any first rotating elements 142. Specifically, the first rotating elements 142 are at least located at 1 / 4, 1 / 2, and 3 / 4 positions diagonally of the first vibrating element 141.

[0194] In some of these embodiments, the first rotating element 142 is a rotating seat.

[0195] At least five second vibration elements 143 are distributed at the bottom of each first vibration element 141, wherein the first vibration element 141 at the bottommost position is not provided with any second vibration element 143.

[0196] The bottom end of the second vibration element 143 is at a certain distance from the inner bottom surface of the culture element 131.

[0197] In some of these embodiments, the second vibrating element 143 is a vibrating rod.

[0198] Each second vibration element 143 has a second rotating element 144 at its end, which is rotatably connected to the first rotating element 142.

[0199] In some of these embodiments, the second rotating element 144 is a rotating shaft.

[0200] The second vibration element 143 has an initial state and an operating state. In the initial state, the second vibration element 143 is arranged parallel to the first vibration element 141 and embedded inside the first vibration element 141, without protruding from the first vibration element 141. In the operating state, the second rotating element 144 rotates relative to the first rotating element 142 so that the second vibration element 143 is arranged perpendicular to the first vibration element 141.

[0201] Furthermore, the first vibration unit 140 also includes a driving element, which is mounted on the bottom of the support element 121 and connected to the first vibration element 141, the second vibration element 143, and the second rotation element 144 respectively, for controlling the operation of the above elements respectively.

[0202] The first vibration element 141 vibrates for a long time, while the second vibration element 143 vibrates intermittently, thereby causing the cell culture medium inside the culture element 131 to flow and improving the culture efficiency.

[0203] The temperature control unit includes a temperature monitoring element and a temperature regulating element. The temperature monitoring element is located inside the enclosure unit 110 and is used to monitor the internal temperature of the enclosure unit 110; the temperature regulating element is located inside the enclosure unit 110 and is used to regulate the internal temperature of the enclosure unit 110.

[0204] Specifically, both the temperature monitoring element and the temperature regulating element are located inside the housing element 111.

[0205] In some of these embodiments, the temperature monitoring element is a temperature sensor.

[0206] In some of these embodiments, the temperature regulating element is a hot air device or a semiconductor temperature control device.

[0207] Furthermore, the culture apparatus 100 also includes a first delivery tube unit 150, a second delivery tube unit 160, and a third delivery tube unit 170. The first delivery tube unit 150 is disposed inside the box unit 110 and connected to the culture unit 130, for providing rinsing solution to the culture unit 130; the second delivery tube unit 160 is disposed inside the box unit 110 and connected to the culture unit 130, for providing recovered cell culture medium to the culture unit 130; and the third delivery tube unit 170 is disposed inside the box unit 110 and connected to the culture unit 130, for outputting the cell culture medium from the culture unit 130 to the outside.

[0208] The first delivery tube unit 150 includes a first input tube element and a plurality of first output tube elements. The first input tube element is disposed inside the housing unit 110; the plurality of first output tube elements are disposed inside the housing unit 110 and connected to the first input tube element and the culture unit 130, respectively. The second delivery tube unit 160 includes a second input tube element and a plurality of output tube elements. The second input tube element is disposed inside the housing unit 110; the plurality of second output tube elements are disposed inside the housing unit 110 and connected to the second input tube element and the culture unit 130, respectively. The third delivery tube unit 170 includes a plurality of third input tube elements and a third output tube element. The plurality of third input tube elements are disposed inside the housing unit 110 and connected to the culture unit 130; the third output tube elements are disposed inside the housing unit 110 and connected to the third input tube elements.

[0209] Specifically, the first input transistor element is connected to the first inlet element; the first output transistor element is connected to the corresponding third inlet element. The second input transistor element is connected to the second inlet element; the second output transistor element is connected to the corresponding fourth inlet element. The third input transistor element is connected to the corresponding second outlet element; the third output transistor element is connected to the first outlet element.

[0210] The number of the first output transistor components is the same as the number of the third inlet transistor components. The number of the second output transistor components is the same as the number of the fourth inlet transistor components.

[0211] Specifically, the usage method of this embodiment is as follows:

[0212] The cell culture medium is placed inside the culture element 131;

[0213] The culture element 131 is placed on the shelf element 122, so that the first limiting element 123 corresponds to and slides with the second limiting element 132;

[0214] The second locking element 133 is aligned with the corresponding first locking element 124, and the first positioning element 125 is aligned with the second positioning element 134. The third locking element 135 is connected to the first positioning element 125 and the second positioning element 134 in sequence to complete the fixation of the culture element 131.

[0215] The second rotating element 144 is controlled to rotate, so that the second vibrating element 143 extends into the interior of the culture element 131;

[0216] The first vibration element 141 is controlled to vibrate the bottom of the culture element 131, and the second vibration element 143 is controlled to intermittently vibrate the inside of the culture element 131.

[0217] After the culture is completed, rinsing solution is delivered into the culture element 131 through the first delivery tube unit 150, and the cell culture medium inside the culture element 131 is delivered to the subsequent separation device through the third delivery tube unit 170.

[0218] The advantages of this invention are that the cell culture medium in the culture unit flows vertically by using the first vibration element to vibrate for a long time, and the cell culture medium in the culture unit flows horizontally in a ripple pattern by using the second vibration element to vibrate intermittently, which improves the culture efficiency and shortens the culture time; the combination of the specially designed first limiting element and the second limiting element effectively suppresses the vertical vibration of the culture element itself.

[0219] Example 2

[0220] like Figure 6As shown, an exosome culture, separation and collection system includes a culture device 100, a separation device 200, a collection device 300, a rinsing device 400, a reflux device 500 and a control device as described in Example 1. The culture device 100 contains cell culture medium for culturing exosomes; the separation device 200 is connected to the culture device 100 and is used to perform multi-stage separation of the cell culture medium delivered by the culture device 100 to obtain exosomes and secondary cell culture medium; the collection device 300 is connected to the separation device 200 and is used to collect the exosomes separated by the separation device 200; the rinsing device 400 is connected to both the culture device 100 and the separation device 200 and is used to provide rinsing solution to both devices, rinsing the cell culture medium from the culture device 100 to the separation device 200 and for separation by the separation device 200, wherein the composition of the rinsing solution is the same as that of the cell culture medium; the reflux device 500 is connected to both the culture device 100 and the separation device 200 and is used to deliver the secondary cell culture medium from the separation device 200 to the culture device 100; and the control device is connected to the culture device 100, the separation device 200, the collection device 300, the rinsing device 400, and the reflux device 500.

[0221] like Figure 7 As shown, the separation device 200 includes a separation unit 210, a vacuum unit 220, an electrode unit 230, and a second vibration unit 240. The separation unit 210 is connected to the culture device 100, the collection device 300, the rinsing device 400, and the reflux device 500, respectively. The vacuum unit 220 is located on both sides of the separation unit 210 and is connected to both the control device and the separation unit 210, respectively, and is used to alternately provide negative pressure to the separation unit 210 under the control of the control device. The electrode unit 230 is located on both sides of the separation unit 210 and is connected to both the control device and the separation unit 210, and is used to alternately generate an electric field inside the separation unit 210 under the control of the control device. The second vibration unit 240 is connected to both the separation unit 210 and the control device, and is used to vibrate the separation unit 210.

[0222] The separation unit 210 includes a first chamber element 211, a second chamber element 212, a third chamber element 213, a first filter element 214, a second filter element 215, a fifth inlet element, a sixth inlet element, a third outlet element, a fourth outlet element, a fifth outlet element, a seventh valve element, an eighth valve element, a ninth valve element, a tenth valve element, and an eleventh valve element. The first chamber element 211 is connected to the culture device 100, the rinsing device 400, and the reflux device 500, respectively. The second chamber element 212 is symmetrically arranged on both sides of the first chamber element 211 and communicates with it. The third chamber element 213 is symmetrically arranged on both sides of the second chamber element 212 and communicates with it, and is also connected to the vacuum unit 220. The first filter element 214 is disposed between the first chamber element 211 and the second chamber element 212. The second filter element 215 is disposed between the second chamber element 212 and the third chamber element 213, and the pore size of the second filter element 215 is smaller than that of the first filter element 214. The fifth inlet element is disposed on one side of the first chamber element 211 and is connected to the culture device 100. 0 connection; the sixth inlet element is located on one side of the first chamber element 211 and connected to the flushing device 400; the third outlet element is located on one side of the first chamber element 211 and connected to the reflux device 500; the fourth outlet element is located on one side of the second chamber element 212 and connected to the collection device 300; the fifth outlet element is located on one side of the third chamber element 213 and connected to the collection device 300; the seventh valve element is located on the fifth inlet element and connected to the control device; the eighth valve element is located on the sixth inlet element and connected to the control device; the ninth valve element is located on the third outlet element and connected to the control device; the tenth valve element is located on the fourth outlet element and connected to the control device; the eleventh valve element is located on the fifth outlet element and connected to the control device.

[0223] Specifically, the fifth inlet element is connected to the second outlet element.

[0224] There are two second chamber elements 212, symmetrically arranged on both sides of the first chamber element 211, and both are connected to the first chamber element 211. There are two third chamber elements 213, respectively arranged on the outside of the corresponding second chamber element 212, and both are connected to the corresponding second chamber element 212.

[0225] The height of the first chamber element 211 is greater than the height of the second chamber element 212. The height of the second chamber element 212 is greater than or equal to the height of the third chamber element 213.

[0226] A first filter element 214 is provided at the connection between the first chamber element 211 and the second chamber element 212. That is, there are two first filter elements 214. A second filter element 215 is provided at the connection between the second chamber element 212 and the third chamber element 213. That is, there are two second filter elements 215.

[0227] The filter pore size of the second filter element 215 is smaller than that of the first filter element 214.

[0228] In some of these embodiments, the first filter element 214 is a filter membrane; the second filter element 215 is a filter membrane.

[0229] The fifth inlet element is located on one side of the top of the first chamber element 211; the sixth inlet element is located on the other side of the top of the first chamber element 211; the third outlet element is located at the bottom of the first chamber element 211; the fourth outlet element is located at the bottom of the second chamber element 212; and the fifth outlet element is located on the side or bottom of the third chamber element 213.

[0230] In some embodiments, the first filter element 214 is also disposed in the fourth outlet element.

[0231] In some embodiments, the fifth inlet element is a fifth liquid inlet, with an interface on its outer side for connecting to a liquid delivery pipe; the sixth inlet element is a sixth liquid inlet, with an interface on its outer side for connecting to a liquid delivery pipe; the third outlet element is a third liquid outlet, with an interface on its outer side for connecting to a liquid delivery pipe; the fourth outlet element is a fourth liquid outlet, with an interface on its outer side for connecting to a liquid delivery pipe; and the fifth outlet element is a fifth liquid outlet, with an interface on its outer side for connecting to a liquid delivery pipe.

[0232] The seventh valve element is located inside the fifth inlet element and is used to control the opening and closing of the fifth inlet element; the eighth valve element is located inside the sixth inlet element and is used to control the opening and closing of the sixth inlet element; the ninth valve element is located inside the third outlet element and is used to control the opening and closing of the third outlet element; the tenth valve element is located inside the fourth outlet element and is used to control the opening and closing of the fourth outlet element; the eleventh valve element is located inside the fifth outlet element and is used to control the opening and closing of the fifth outlet element.

[0233] In some of these embodiments, the fourth valve element is a fourth solenoid valve; the fifth valve element is a fifth solenoid valve; the ninth valve element is a ninth solenoid valve; the tenth valve element is a tenth solenoid valve; and the eleventh valve element is an eleventh solenoid valve.

[0234] Vacuum unit 220 includes a first vacuum element 221 and a second vacuum element 222. The first vacuum element 221 is disposed on one side of separation unit 210 and is connected to separation unit 210 and control device respectively; the second vacuum element 222 is disposed on the other side of separation unit 210 and is connected to separation unit 210 and control device respectively; wherein the first vacuum element 221 and the second vacuum element 222 alternately provide negative pressure to separation unit 210.

[0235] Specifically, the first vacuum element 221 is connected to the third chamber element 213 on one side; the second vacuum element 222 is connected to the third chamber element 213 on the other side. When the first vacuum element 221 provides negative pressure to the third chamber element 213 and the second chamber element 212 on one side, the second vacuum element 222 does not provide negative pressure to the third chamber element 213 and the second chamber element 212 on the other side.

[0236] In some of these embodiments, the first vacuum element 221 and the second vacuum element 222 are negative pressure generating devices, such as vacuum pumps.

[0237] Electrode unit 230 includes a first electrode element 231 and a second electrode element 232. The first electrode element 231 is disposed on one side of the interior of separation unit 210 and is connected to separation unit 210 and control device respectively; the second electrode element 232 is disposed on the other side of the interior of separation unit 210 and is connected to separation unit 210 and control device respectively; wherein the first electrode element 231 and the second electrode element 232 alternately generate an electric field inside separation unit 210.

[0238] Specifically, the first electrode element 231 is disposed inside the third chamber element 213 on one side; the second electrode element 232 is disposed inside the third chamber element 213 on the other side.

[0239] In some embodiments, when the first electrode element 231 is energized and generates an electric field, the second electrode element 232 is not energized; when the second electrode element 232 is energized and generates an electric field, the first electrode element 231 is not energized.

[0240] In some embodiments, when both the first electrode element 231 and the second electrode element 232 are energized, the first electrode element 231 is the positive electrode and the second electrode element 232 is the negative electrode, or the first electrode element 231 is the negative electrode and the second electrode element 232 is the positive electrode, and the first electrode element 231 and the second electrode element 232 alternately become the positive and negative electrodes.

[0241] When the first vacuum element 221 is working, the first electrode element 231 generates an electric field or is the positive electrode; when the second vacuum element 222 is working, the second electrode element 232 generates an electric field or is the positive electrode. That is, through the combined action of the vacuum element and the electrode element, the cell culture medium inside the separation unit 210 flows to one side.

[0242] In some embodiments, the first electrode element 231 and the second electrode element 232 are electrodes, or the first electrode element 231 and the second electrode element 232 are electromagnetic coils.

[0243] The second vibration unit 240 includes a third vibration element 241 and a fourth vibration element 242. The third vibration element 241 is disposed on the side of the separation unit 210 and connected to the control device; the fourth vibration element 242 is disposed on the side of the separation unit 210 and connected to the control device.

[0244] Specifically, the third vibration element 241 is disposed on the side of the second chamber element 212 and the third chamber element 213 on one side; the fourth vibration element 242 is disposed on the side of the second chamber element 212 and the third chamber element 213 on the other side.

[0245] When the first vacuum element 221 provides negative pressure, the third vibration element 241 located on the same side vibrates, causing the first filter element 214 and the second filter element 215 on the same side to vibrate, thereby preventing the first filter element 214 and the second filter element 215 from becoming clogged.

[0246] In some of these embodiments, the third vibration element 241 and the fourth vibration element 242 are oscillators or vibration motors.

[0247] In some embodiments, the third vibration element 241 and the fourth vibration element 242 may also be directly disposed on the first filter element 214 and the second filter element 215.

[0248] Furthermore, the separation device 200 also includes a fourth delivery pipe unit 250, a fifth delivery pipe unit 260, a sixth delivery pipe unit 270, and a seventh delivery pipe unit 280. The fourth delivery pipe unit 250 is connected to the culture device 100 and the separation unit 210, respectively; the fifth delivery pipe unit 260 is connected to the rinsing device 400 and the separation unit 210, respectively; the sixth delivery pipe unit 270 is connected to the separation unit 210 and the reflux device 500, respectively; and the seventh delivery pipe unit 280 is connected to the separation unit 210 and the collection device 300, respectively.

[0249] Specifically, the fourth delivery pipe unit 250 is connected to the second outlet element and the fifth inlet element respectively; the fifth delivery pipe unit 260 is connected to the sixth inlet element; the sixth delivery pipe unit 270 is connected to the third outlet element; and the seventh delivery pipe unit 280 is connected to the fourth outlet element and the fifth outlet element respectively.

[0250] like Figure 8 As shown, the collection device 300 includes a first collection unit 310 and a second collection unit 320. The first collection unit 310 is connected to the separation device 200 and the control device, respectively, and is used to collect exosomes obtained by primary separation by the separation device 200; the second collection unit 320 is connected to the separation device 200 and the control device, respectively, and is used to collect exosomes obtained by multi-stage separation by the separation device 200.

[0251] The first collection unit 310 includes a first liquid conveying element 311, at least one first storage element 312, at least one seventh inlet element, at least one twelfth valve element, and at least one first liquid level monitoring element. The first liquid conveying element 311 is connected to both the separation device 200 and the control device, and is used to convey exosomes obtained through primary separation by the separation device 200. The first storage element 312 is connected to the first liquid conveying element 311 and is used to collect the exosomes obtained through primary separation by the separation device 200. The seventh inlet element is disposed in the first storage element 312 and connected to the first liquid conveying element 311. The twelfth valve element is disposed in the seventh inlet element and connected to the control device. The first liquid level monitoring element is disposed inside the first storage element 312 and connected to the control device.

[0252] Specifically, the first liquid delivery element 311 is connected to the fourth outlet element through the seventh delivery pipe unit 280.

[0253] When there are multiple first storage elements 312, the first liquid delivery element 311 is connected to a plurality of first storage elements 312 respectively.

[0254] The first storage element 312, the seventh inlet element, the twelfth valve element, and the first liquid level monitoring element correspond one-to-one. The twelfth valve element is located inside the seventh inlet element.

[0255] In some of these embodiments, the first liquid delivery element 311 is a first water pump; the first storage element 312 is a first liquid storage tank; the seventh inlet element is a seventh liquid inlet with an interface on its outer side for connecting to a liquid delivery pipe; the twelfth valve element is a twelfth solenoid valve; and the first liquid level monitoring element is a first liquid level sensor.

[0256] The second collection unit 320 includes a second liquid conveying element 321, at least one second storage element 322, at least one eighth inlet element, at least one thirteenth valve element, and at least one second liquid level monitoring element. The second liquid conveying element 321 is connected to both the separation device 200 and the control device, and is used to convey exosomes obtained through multi-stage separation by the separation device 200. The second storage element 322 is connected to the second liquid conveying element 321 and is used to collect the exosomes obtained through multi-stage separation by the separation device 200. The eighth inlet element is disposed in the second storage element 322 and connected to the second liquid conveying element 321. The thirteenth valve element is disposed in the eighth inlet element and connected to the control device. The second liquid level monitoring element is disposed inside the second storage element 322 and connected to the control device.

[0257] Specifically, the second liquid delivery element 321 is connected to the fifth outlet element via the seventh delivery pipe unit 280.

[0258] When there are multiple second storage elements 322, the second liquid delivery element 321 is connected to a plurality of second storage elements 322 respectively.

[0259] The second storage element 322, the eighth inlet element, the thirteenth valve element, and the second liquid level monitoring element correspond one-to-one. The thirteenth valve element is located inside the eighth inlet element.

[0260] In some embodiments, the second liquid delivery element 321 is a second water pump; the second storage element 322 is a second liquid storage tank; the eighth inlet element is an eighth liquid inlet with an interface on its outer side for connecting to a liquid delivery pipe; the thirteenth valve element is a thirteenth solenoid valve; and the second liquid level monitoring element is a second liquid level sensor.

[0261] Furthermore, the collection device 300 also includes an eighth conveying pipe unit 330 and a ninth conveying pipe unit 340. The eighth conveying pipe unit 330 is connected to the separation device 200 and the first collection unit 310, respectively; the ninth conveying pipe unit 340 is connected to the separation device 200 and the second collection unit 320, respectively.

[0262] Specifically, the eighth delivery pipe unit 330 is connected to the first liquid delivery element 311 and at least one first storage element 312, respectively; the ninth delivery pipe unit 340 is connected to the second liquid delivery element 321 and at least one second storage element 322, respectively. More specifically, the eighth delivery pipe unit 330 is connected to the seventh inlet element; the ninth delivery pipe unit 340 is connected to the eighth inlet element.

[0263] like Figure 9As shown, the rinsing device 400 includes a first liquid storage unit 410, a first rinsing unit 420, and a second rinsing unit 430. The first liquid storage unit 410 is disposed on one side of the culture device 100 and is used to store rinsing solution. The first rinsing unit 420 is connected to the first liquid storage unit 410, the culture device 100, and the control device, respectively, and is used to provide rinsing solution to the culture device 100 to rinse the cell culture medium in the culture device 100 to the separation device 200. The second rinsing unit 430 is connected to the first liquid storage unit 410, the separation device 200, and the control device, respectively, and is used to provide rinsing solution to the separation device 200 for separation.

[0264] The first liquid storage unit 410 includes a third storage element 411, a ninth inlet element, a sixth outlet element, a seventh outlet element, a fourteenth valve element, a fifteenth valve element, and a third liquid level monitoring element. The third storage element 411 is disposed on one side of the culture device 100 and is used to store rinsing liquid; the ninth inlet element is disposed on one side of the third storage element 411 and is used to replenish the rinsing liquid inside the third storage element 411; the sixth outlet element is disposed on one side of the third storage element 411 and is connected to the first rinsing unit 420; the seventh outlet element is disposed on one side of the third storage element 411 and is connected to the second rinsing unit 430; the fourteenth valve element is disposed on the sixth outlet element and is connected to a control device; the fifteenth valve element is disposed on the seventh outlet element and is connected to a control device; and the third liquid level monitoring element is disposed inside the third storage element 411 and is connected to the control device.

[0265] The ninth inlet element is located on top of the third storage element 411; the sixth outlet element is located on one side of the bottom of the third storage element 411; the seventh outlet element is located on the other side of the bottom of the third storage element 411; the fourteenth valve element is located inside the sixth outlet element; and the fifteenth valve element is located inside the seventh outlet element.

[0266] In some embodiments, the third storage element 411 is a third liquid storage tank; the ninth inlet element is a ninth liquid inlet, with an interface on its outer side for connecting to a liquid delivery pipe; the sixth outlet element is a sixth liquid outlet, with an interface on its outer side for connecting to a liquid delivery pipe; the seventh outlet element is a seventh liquid outlet, with an interface on its outer side for connecting to a liquid delivery pipe; the fourteenth valve element is a fourteenth solenoid valve; the fifteenth valve element is a fifteenth solenoid valve; and the third liquid level monitoring element is a third liquid level sensor.

[0267] The first rinsing unit 420 includes a third liquid delivery element 421 and a first flow monitoring element. The third liquid delivery element 421 is connected to the first liquid storage unit 410, the culture device 100, and the control device. The first flow monitoring element is disposed on the third liquid delivery element 421 and is connected to the control device.

[0268] Specifically, the third liquid conveying element 421 is connected to the first liquid storage unit 410 and the tank unit 110, respectively. More specifically, the third liquid conveying element 421 is connected to the third storage element 411 and the tank element 111, respectively. More specifically, the third liquid conveying element 421 is connected to the seventh outlet element and the first inlet element, respectively.

[0269] In some of these embodiments, the third liquid delivery element 421 is a third water pump; and the first flow monitoring element is a first flow meter.

[0270] The second flushing unit 430 includes a fourth liquid delivery element 431 and a second flow monitoring element. The fourth liquid delivery element 431 is connected to the first liquid storage unit 410, the separation device 200, and the control device, respectively; the second flow monitoring element is disposed on the fourth liquid delivery element 431 and is connected to the control device.

[0271] Specifically, the fourth liquid conveying element 431 is connected to the first liquid storage unit 410 and the separation unit 210, respectively. More specifically, the fourth liquid conveying element 431 is connected to the eighth outlet element and the sixth inlet element, respectively.

[0272] In some of these embodiments, the fourth liquid delivery element 431 is a fourth water pump; and the second flow monitoring element is a second flow meter.

[0273] Furthermore, the rinsing device 400 also includes a tenth delivery pipe unit 440 and an eleventh delivery pipe unit 450. The tenth delivery pipe unit 440 is connected to the first liquid storage unit 410 and the culture device 100, respectively; the eleventh delivery pipe unit 450 is connected to the first liquid storage unit 410 and the separation device 200, respectively.

[0274] Specifically, the tenth delivery pipe unit 440 is connected to the first liquid storage unit 410 and the tank unit 110, respectively; the eleventh delivery pipe unit 450 is connected to the first liquid storage unit 410 and the separation unit 210, respectively. More specifically, the tenth delivery pipe unit 440 is connected to the seventh outlet element, the third liquid delivery element 421, and the first inlet element, respectively; the eleventh delivery pipe unit 450 is connected to the eighth outlet element, the fourth liquid delivery element 431, and the sixth inlet element, respectively.

[0275] In some embodiments, the eleventh delivery pipe unit 450 and the fifth delivery pipe unit 260 are the same liquid delivery pipe.

[0276] like Figure 10 As shown, the reflux device 500 includes a filtration unit 510, a second liquid storage unit 520, a third liquid storage unit 530, and a reflux unit 540. The filtration unit 510 is connected to the separation device 200; the second liquid storage unit 520 is disposed on one side of the filtration unit 510 and connected to it; the third liquid storage unit 530 is disposed on one side of the filtration unit 510 and connected to it; the reflux unit 540 is connected to the culture device 100, the second liquid storage unit 520, and the control device, respectively, and is used to reflux the secondary cell culture medium obtained by the separation device 200 back to the culture device 100.

[0277] The filtration unit 510 includes a fifth liquid delivery element 511, a fourth storage element 512, an electrostatic adsorption element 513, a third filtration element 514, a tenth inlet element, an eighth outlet element, a ninth outlet element, a sixteenth valve element, a seventeenth valve element, and an eighteenth valve element. The fifth liquid conveying element 511 is connected to the separation device 200 and the control device, respectively; the fourth storage element 512 is connected to the fifth liquid conveying element 511, the second liquid storage unit 520, and the third liquid storage unit 530, respectively; the electrostatic adsorption element 513 is disposed inside the fourth storage element 512 and is connected to the control device; the third filter element 514 is disposed inside the fourth storage element 512 and is located downstream of the electrostatic adsorption element 513; the tenth inlet element is disposed on one side of the fourth storage element 512 and is connected to the fifth liquid conveying element 511; the eighth outlet element is disposed on one side of the fourth storage element 512, is located downstream of the third filter element 514, and is connected to the second liquid storage unit 520; the ninth outlet element is disposed on one side of the fourth storage element 512, is located upstream of the third filter element 514, and is connected to the third liquid storage unit 530; the sixteenth valve element is disposed on the tenth inlet element and is connected to the control device; the seventeenth valve element is disposed on the eighth outlet element and is connected to the control device; and the eighteenth valve element is disposed on the ninth outlet element and is connected to the control device.

[0278] Specifically, the fifth liquid delivery element 511 is connected to the first chamber element 211. More specifically, the fifth liquid delivery element 511 is connected to the third outlet element.

[0279] When energized, the electrostatic adsorption element 513 electrostatically adsorbs the cell culture medium located inside the fourth storage element 512 to adsorb larger substances; when the electrostatic adsorption element 513 is not energized, it does not adsorb any substances and releases the adsorbed substances.

[0280] An electrostatic adsorption element 513 is disposed in the upper middle / front middle part of the fourth storage element 512; a third filter element 514 is disposed in the middle of the fourth storage element 512; a tenth inlet element is disposed at the top of the fourth storage element 512; an eighth outlet element is disposed at the bottom of the fourth storage element 512; a ninth outlet element is disposed on the side of the fourth storage element 512; a sixteenth valve element is disposed inside the tenth inlet element; a seventeenth valve element is disposed inside the eighth outlet element; and an eighteenth valve element is disposed inside the ninth outlet element.

[0281] In some embodiments, the pore size of the third filter element 514 is smaller than that of the second filter element 215.

[0282] In some embodiments, the fifth liquid delivery element 511 is a fifth water pump; the fourth storage element 512 is a fourth liquid storage tank; the electrostatic adsorption element 513 is an electrostatic adsorption membrane; the third filter element 514 is a filter membrane; the tenth inlet element is a tenth liquid inlet, with an interface on its outer side for connecting to a liquid delivery pipe; the eighth outlet element is an eighth liquid outlet, with an interface on its outer side for connecting to a liquid delivery pipe; the ninth outlet element is a ninth liquid outlet, with an interface on its outer side for connecting to a liquid delivery pipe; the sixteenth valve element is a sixteenth solenoid valve; the seventeenth valve element is a seventeenth solenoid valve; and the eighteenth valve element is an eighteenth solenoid valve.

[0283] The second liquid storage unit 520 includes a sixth liquid delivery element 521, a fifth storage element 522, an eleventh inlet element, a twelfth inlet element, a tenth outlet element, a nineteenth valve element, and a twentieth valve element. The sixth liquid delivery element 521 is connected to the filtration unit 510 and the control device. The fifth storage element 522 is connected to the sixth liquid delivery element 521 and the reflux unit 540. The eleventh inlet element is located on one side of the fifth storage element 522 and connected to the sixth liquid delivery element 521. The twelfth inlet element is located on one side of the fifth storage element 522 and is used to replenish cultured cells into the fifth storage element 522. The tenth outlet element is located on one side of the fifth storage element 522 and connected to the reflux unit 540. The nineteenth valve element is located on the eleventh inlet element and connected to the control device. The twentieth valve element is located on the tenth outlet element and connected to the control device.

[0284] Specifically, the sixth liquid conveying element 521 is connected to the fourth storage element 512. More specifically, the sixth liquid conveying element 521 is connected to the eighth outlet element.

[0285] The eleventh inlet element is located on one side of the top of the fifth storage element 522; the twelfth inlet element is located on the other side of the top of the fifth storage element 522; the tenth outlet element is located at the bottom of the fifth storage element 522; the nineteenth valve element is located inside the eleventh inlet element; and the twentieth valve element is located inside the tenth outlet element.

[0286] In some embodiments, the sixth liquid delivery element 521 is a sixth water pump; the fifth storage element 522 is a fifth liquid storage tank; the eleventh inlet element is an eleventh liquid inlet with an interface on its outer side for connecting to a liquid delivery pipe; the twelfth inlet element is a twelfth liquid inlet with an interface on its outer side for connecting to a liquid delivery pipe; the tenth outlet element is a tenth liquid outlet with an interface on its outer side for connecting to a liquid delivery pipe; the nineteenth valve element is a nineteenth solenoid valve; and the twentieth valve element is a twentieth solenoid valve.

[0287] The third liquid storage unit 530 includes a seventh liquid delivery element 531, a sixth storage element 532, a thirteenth inlet element, and a twenty-first valve element. The seventh liquid delivery element 531 is connected to the filter unit 510 and the control device; the sixth storage element 532 is connected to the seventh liquid delivery element 531; the thirteenth inlet element is located on the sixth storage element 532 and connected to the seventh liquid delivery element 531; and the twenty-first valve element is located on the twelfth inlet element and connected to the control device.

[0288] Specifically, the seventh liquid conveying element 531 is connected to the fourth storage element 512. More specifically, the seventh liquid conveying element 531 is connected to the ninth outlet element.

[0289] The thirteenth inlet element is located on one side of the top of the sixth storage element 532; the twenty-first valve element is located inside the thirteenth inlet element.

[0290] In some embodiments, the seventh liquid delivery element 531 is a seventh water pump; the sixth storage element 532 is a sixth liquid storage tank; the thirteenth inlet element is a thirteenth liquid inlet with an interface on its outer side for connecting to a liquid delivery pipe; and the twenty-first valve element is a twenty-first solenoid valve.

[0291] The reflux unit 540 includes an eighth liquid delivery element 541 and a third flow monitoring element. The eighth liquid delivery element 541 is connected to the second liquid storage unit 520, the culture device 100, and the control device. The third flow monitoring element is disposed on the eighth liquid delivery element 541 and is connected to the control device.

[0292] Specifically, the eighth liquid conveying element 541 is connected to the second liquid storage unit 520 and the tank unit 110, respectively. More specifically, the eighth liquid conveying element 541 is connected to the fifth storage element 522 and the tank element 111, respectively. More specifically, the eighth liquid conveying element 541 is connected to the tenth outlet element and the second inlet element, respectively.

[0293] In some of these embodiments, the eighth liquid delivery element 541 is an eighth water pump; the third flow monitoring element is a third flow meter.

[0294] Furthermore, the reflux device 500 also includes a twelfth delivery pipe unit 550, a thirteenth delivery pipe unit 560, a fourteenth delivery pipe unit 570, and a fifteenth delivery pipe unit 580. The twelfth delivery pipe unit 550 is connected to the separation device 200 and the filtration unit 510, respectively; the thirteenth delivery pipe unit 560 is connected to the filtration unit 510 and the second liquid storage unit 520, respectively; the fourteenth delivery pipe unit 570 is connected to the filtration unit 510 and the third liquid storage unit 530, respectively; and the fifteenth delivery pipe unit 580 is connected to the second liquid storage unit 520 and the culture device 100, respectively.

[0295] Specifically, the twelfth delivery pipe unit 550 is connected to the separation unit 210 and the filtration unit 510, respectively; the fifteenth delivery pipe unit 580 is connected to the second storage unit 520 and the tank unit 110, respectively. More specifically, the twelfth delivery pipe unit 550 is connected to the first chamber element 211 and the fourth storage element 512, respectively; the thirteenth delivery pipe unit 560 is connected to the fourth storage element 512 and the fifth storage element 522, respectively; the fourteenth delivery pipe unit 570 is connected to the fourth storage element 512 and the sixth storage element 532, respectively; and the fifteenth delivery pipe unit 580 is connected to the fifth storage element 522 and the tank element 111, respectively. More specifically, the twelfth delivery pipe unit 550 is connected to the third outlet element and the tenth inlet element, respectively; the thirteenth delivery pipe unit 560 is connected to the eighth outlet element and the eleventh inlet element, respectively; the fourteenth delivery pipe unit 570 is connected to the ninth outlet element and the thirteenth inlet element, respectively; and the fifteenth delivery pipe unit 580 is connected to the tenth outlet element and the second inlet element, respectively.

[0296] In some of these embodiments, the twelfth delivery pipe unit 550 and the sixth delivery pipe unit 270 are the same liquid delivery pipe.

[0297] The control device is a central control device, which can be set on one side of the culture device 100 or in the central control room.

[0298] The advantages of this invention are: it utilizes a culture device for large-scale culture of exosomes, improving culture efficiency; it uses a separation and collection device for multi-stage separation and collection of exosomes, obtaining exosomes of different sizes to meet different needs; it utilizes a vacuum unit and an electrode unit working together to further improve separation efficiency; and it uses a reflux device to filter and adsorb the secondary cell culture medium separated by the separation unit and reflux it, further reducing the cost of exosome culture.

[0299] Example 3

[0300] A method for culturing, isolating, and collecting exosomes, applied to the exosome culture, isolating, and collecting system described in Example 2, includes:

[0301] Step S702: Place cell culture medium into culture device 100 for culturing exosomes;

[0302] Step S704: When the culture time of the culture device 100 reaches the preset culture time threshold, the control device controls the rinsing device 400 to provide rinsing solution to the interior of the culture device 100 so that all the cell culture medium in the culture device 100 enters the separation device 200. The composition of the rinsing solution is the same as that of the cell culture medium.

[0303] Step S706: The control device controls the separation device 200 to perform multi-stage separation of the cell culture medium to obtain exosomes and secondary cell culture medium. During the multi-stage separation of the separation device 200, the control device controls the rinsing device 400 to provide rinsing solution to the interior of the separation device 200.

[0304] Step S708: After the separation device 200 has completed multi-stage separation, the control device controls the collection device 300 to collect the exosomes obtained by the separation device 200.

[0305] Step S710: After the separation device 200 has completed the primary separation, the control device controls the reflux device 500 to reflux the secondary cell culture medium obtained by the primary separation in the separation device 200 back to the culture device 100 for use in culturing exosomes.

[0306] Specifically, methods for culturing, isolating, and collecting exosomes include:

[0307] Step S7022: Place cell culture medium into the culture unit 130 of the culture device 100 for culturing exosomes;

[0308] Step S7024: The control device controls the first vibration unit 140 of the culture device 100 to vibrate the culture unit 130 to increase the culture rate;

[0309] Step S7042: When the culture time of culture unit 130 reaches the preset culture time threshold, the control device controls the first rinsing unit 420 of rinsing device 400 to provide rinsing solution to the interior of culture unit 130 so that the cell culture medium of culture unit 130 enters the separation unit 210 of separation device 200.

[0310] Step S7062: The control device controls the vacuum unit 220, electrode unit 230, and second vibration unit 240 of the separation device 200 to perform multi-stage separation of the cell culture medium in the separation unit 210 to obtain exosomes and secondary cell culture medium. In the case of multi-stage separation in the separation unit 210, the control device controls the second rinsing unit 430 of the rinsing device 400 to provide rinsing solution to the separation unit 210.

[0311] Step S7082: After the separation unit 210 has completed multi-stage separation, the control device controls the first collection unit 310 of the collection device 300 to collect the exosomes separated in the first stage, and the second collection unit 320 of the collection device 300 to collect the exosomes separated in the multi-stage separation.

[0312] Step S7102: After the separation unit 210 has completed the primary separation, the control device controls the filtration unit 510 of the reflux device 500 to filter the secondary cell culture medium.

[0313] Step S7104: The control device controls the second liquid storage unit 520 of the reflux device 500 to collect the secondary cell culture medium filtered by the filtration unit 510.

[0314] Step S7106: The control device controls the third storage unit 530 of the reflux device 500 to collect the secondary cell culture medium that has not been filtered by the filtration unit 510.

[0315] Step S7108: The control device controls the reflux unit 540 of the reflux device 500 to reflux the secondary cell culture medium collected in the second storage unit 520 back to the culture unit 130.

[0316] More specifically, exosome culture isolation and collection methods include:

[0317] (a) Exosome culture

[0318] The cell culture medium is placed inside the culture element 131;

[0319] The culture element 131 is placed on the shelf element 122, so that the first limiting element 123 corresponds to and slides with the second limiting element 132;

[0320] The second locking element 133 is aligned with the corresponding first locking element 124, and the first positioning element 125 is aligned with the second positioning element 134. The third locking element 135 is connected to the first positioning element 125 and the second positioning element 134 in sequence to complete the fixation of the culture element 131.

[0321] The control device controls the second rotating element 144 to rotate, so that the second vibrating element 143 extends into the interior of the culture element 131;

[0322] The control device controls the first vibration element 141 to vibrate the bottom of the culture element 131, and the control device controls the second vibration element 143 to vibrate the inside of the culture element 131 intermittently.

[0323] After the exosome culture is completed, the control device controls the first vibration element 141 and the second vibration element 143 to stop working;

[0324] (II) Exosome isolation

[0325] The control device controls the third liquid delivery element 421 to provide flushing fluid to the interior of the culture element 131, so as to flush the cell culture medium of the culture element 131 into the first chamber element 211;

[0326] After the cell culture medium in the culture element 131 has completely entered the interior of the first chamber element 211, the control device controls the fourth liquid delivery element 431 to provide flushing fluid to the interior of the first chamber element 211.

[0327] The control device controls the first vacuum element 221 and the second vacuum element 222 to alternately provide negative pressure to the first chamber element 211, and the first electrode element 231 and the second electrode element 232 to alternately generate electric fields with different directions, so that the cell culture medium inside the first chamber element 211 alternately enters the second chamber element 212 and the third chamber element 213, thereby filtering the cell culture medium through the first filter element 214 and the second filter element 215, thereby obtaining exosomes obtained by primary separation (first exosomes) and exosomes obtained by multi-stage separation (second exosomes).

[0328] During the exosome separation process, the control device also controls the third vibration element 241 and the fourth vibration element 242 to vibrate in order to improve the separation efficiency.

[0329] After the exosomes are separated, the control device controls the first liquid delivery element 311 to deliver the first exosomes to the first storage element 312, and controls the second liquid delivery element 321 to deliver the second exosomes to the second storage element 322.

[0330] (iii) Cell culture medium reflux

[0331] The control device controls the fifth liquid delivery element 511 to deliver the secondary cell culture medium of the first chamber element 211 to the fourth storage element 512;

[0332] The control device controls the sixth liquid delivery element 521 to deliver the secondary cell culture medium filtered by the electrostatic adsorption element 513 and the third filter element 514 to the fifth storage element 522, and the control device controls the seventh liquid delivery element 531 to deliver the secondary cell culture medium that has not been filtered by the third filter element 514 to the sixth storage element 532.

[0333] The materials required for exosome culture are delivered to the fourth storage element 512;

[0334] The control device controls the eighth liquid delivery element 541 to deliver the cell culture medium from the fourth storage element 512 to the culture element 131.

[0335] The above description is merely a preferred embodiment of the present invention and does not limit the implementation and protection scope of the present invention. Those skilled in the art should realize that any equivalent substitutions and obvious changes made based on the description and illustrations of the present invention should be included within the protection scope of the present invention.

Claims

1. A culture device for an exosome culture separation collection system, characterized by, The application relates to a cell culture device. The device comprises: a box unit placed on a horizontal plane; a support unit arranged in the interior of the box unit; a culture unit detachably arranged in the interior of the support unit; a first vibration unit arranged in the support unit and connected with the culture unit, used for vibrating the culture unit; a temperature control unit arranged in the interior of the box unit, used for adjusting the interior temperature of the box unit; wherein the support unit comprises: a support element arranged in the interior of the box unit; a plurality of shelf plate elements arranged in the interior of the support element in the vertical direction of the support element, the shelf plate elements being provided with the first vibration unit; wherein the culture unit comprises: a plurality of culture elements arranged in the interior of the support unit in the vertical direction of the support unit, detachably connected with the support unit, and arranged on the upper part of the corresponding shelf plate elements, the culture elements being culture frames with open tops; wherein the first vibration unit comprises: a plurality of first vibration elements arranged in the interior of the support unit in the vertical direction of the support unit, used for vibrating the bottom of the culture unit; a plurality of first rotating elements arranged on the bottom of the first vibration elements, wherein the first vibration element at the bottom is not provided with a first rotating element; a plurality of second vibration elements arranged on the bottom of the first vibration elements, the bottom end of the second vibration element being a certain distance from the inner bottom surface of the culture element, used for intermittently vibrating the interior of the culture unit so that the cell culture solution of the culture unit flows in the horizontal direction and presents a ripple pattern, wherein the first vibration element at the bottom is not provided with a second vibration element; a plurality of second rotating elements arranged on the end of the second vibration element and connected with the corresponding first rotating element; 2. The culture device of claim 1, wherein wherein the second vibration element has an initial state and a working state; in the case that the second vibration element is in the initial state, the second vibration element is arranged parallel to the first vibration element, embedded in the interior of the first vibration element, and does not protrude from the first vibration element; in the case that the second vibration element is in the working state, the second rotating element rotates relative to the first rotating element, so that the second vibration element is arranged perpendicular to the first vibration element. The box unit comprises: a box element placed on a horizontal plane; a buffer element arranged on the bottom of the box element, used for reducing the impact of external vibration; and / or The support unit further comprises: a plurality of first limiting elements symmetrically arranged on the inner side walls of the left and right sides of the support element, used for detachably connecting with the culture unit; a plurality of first locking elements arranged on the front and back sides of the corresponding shelf plate elements, used for detachably connecting with the culture unit; a plurality of first positioning elements connected with the corresponding first locking elements, used for detachably connecting with the culture unit; and / or The culture unit further comprises: A plurality of second limiting elements are arranged on the left and right sides of the corresponding culture element and are detachably connected with the support unit. A plurality of second locking elements are arranged on the front and back sides of the corresponding culture element and are detachably connected with the support unit. A plurality of second positioning elements are arranged on the front and back sides of the corresponding culture element and are detachably connected with the second locking element. A plurality of third locking elements are detachably connected with the corresponding second locking element, the corresponding second positioning element, and the corresponding second locking element and the support unit; and / or The temperature control unit comprises: A temperature monitoring element arranged in the interior of the box unit for monitoring the interior temperature of the box unit; A temperature adjusting element arranged in the interior of the box unit for adjusting the interior temperature of the box unit.

3. An exosome culture separation collection system, characterized by, Comprise: The culture device of claim 1 or 2, wherein the interior of the culture device is provided with a cell culture solution for culturing exosomes; A separation device connected with the culture device for multi-stage separation of the cell culture solution delivered by the culture device to obtain exosomes and secondary cell culture solution; A collection device connected with the separation device for collecting the exosomes separated by the separation device; A flushing device connected with the culture device and the separation device, respectively, for providing flushing liquid to the culture device and the separation device, respectively, for flushing the cell culture solution of the culture device to the separation device, and for use in separation by the separation device, wherein the composition of the flushing liquid is the same as that of the cell culture solution; A reflux device connected with the culture device and the separation device, respectively, for delivering the secondary cell culture solution of the separation device to the culture device; A control device connected with the culture device, the separation device, the collection device, the flushing device, and the reflux device, respectively.

4. The exosome culture separation collection system according to claim 3, characterized by, The separation device comprises: A separation unit connected with the culture device, the collection device, the flushing device, and the reflux device, respectively; A vacuum unit arranged on both sides of the separation unit and connected with the control device and the separation unit, respectively, for alternately providing negative pressure to the separation unit under the control of the control device; An electrode unit arranged on both sides of the separation unit and connected with the control device and the separation unit, respectively, for alternately generating an electric field in the interior of the separation unit under the control of the control device; A second vibration unit connected with the separation unit and the control device, respectively, for vibrating the separation unit; Wherein, in the case that the vacuum unit provides negative pressure to one side of the separation unit, the vacuum unit does not provide negative pressure to the other side of the separation unit; and / or The collection device comprises: A first collection unit connected with the separation device and the control device, respectively, for collecting exosomes obtained by one-stage separation by the separation device; A second collection unit connected with the separation device and the control device, respectively, for collecting exosomes obtained by multi-stage separation by the separation device; and / or The flushing device comprises: The first liquid storage unit is arranged on one side of the culture device and is used for storing the flushing liquid. The first flushing unit is connected with the first liquid storage unit, the culture device and the control device, respectively, and is used for providing the flushing liquid to the culture device to flush the cell culture liquid of the culture device to the separation device. The second flushing unit is connected with the first liquid storage unit, the separation device and the control device, respectively, and is used for providing the flushing liquid to the separation device for separation. The reflux device comprises: The filtration unit is connected with the separation device. The second liquid storage unit is arranged on one side of the filtration unit and is connected with the filtration unit. The third liquid storage unit is arranged on one side of the filtration unit and is connected with the filtration unit. The reflux unit is connected with the culture device, the second liquid storage unit and the control device, respectively, and is used for refluxing the secondary cell culture liquid obtained through the separation of the separation device to the culture device.

5. The exosome culture separation collection system of claim 4, wherein, The separation unit comprises: The first chamber element is connected with the culture device, the flushing device and the reflux device, respectively. The second chamber element is symmetrically arranged on two sides of the first chamber element and is in communication with the first chamber element. The third chamber element is symmetrically arranged on two sides of the second chamber element, is in communication with the second chamber element, and is in communication with the vacuum unit, respectively. The first filter element is arranged between the first chamber element and the second chamber element. The second filter element is arranged between the second chamber element and the third chamber element, and the pore size of the second filter element is smaller than that of the first filter element. The vacuum unit comprises: The first vacuum element is arranged on one side of the separation unit and is connected with the separation unit and the control device, respectively. The second vacuum element is arranged on the other side of the separation unit and is connected with the separation unit and the control device, respectively. The first vacuum element and the second vacuum element alternately provide negative pressure to the separation unit. The electrode unit comprises: The first electrode element is arranged on one side of the inside of the separation unit and is connected with the separation unit and the control device, respectively. The second electrode element is arranged on the other side of the inside of the separation unit and is connected with the separation unit and the control device, respectively. The first electrode element and the second electrode element alternately generate an electric field in the inside of the separation unit. The second vibration unit comprises: The third vibration element is arranged on the side of the separation unit and is connected with the control device. The fourth vibration element is arranged on the side of the separation unit and is connected with the control device.

6. The exosome culture separation collection system of claim 4, wherein, The first collection unit comprises: The first liquid conveying element is connected with the separation device and the control device, respectively, and is used for conveying the exosomes obtained through the primary separation of the separation device. The at least one first storage element is connected with the first liquid conveying element and is used for collecting the exosomes obtained through the primary separation of the separation device. At least one first liquid level monitoring element is arranged inside the first storage element and connected with the control device; and / or The second collection unit comprises: A second liquid delivery element connected with the separation device and the control device, for delivering the exosomes obtained through multi-stage separation by the separation device; At least one second storage element connected with the second liquid delivery element, for collecting the exosomes obtained through multi-stage separation by the separation device; At least one second liquid level monitoring element arranged inside the second storage element and connected with the control device.

7. The exosome culture separation collection system of claim 4, wherein, The first liquid storage unit comprises: A third storage element arranged on one side of the culture device, for storing the flushing liquid; A third liquid level monitoring element arranged inside the third storage element and connected with the control device; and / or The first flushing unit comprises: A third liquid delivery element connected with the first liquid storage unit, the culture device and the control device; A first flow monitoring element arranged on the third liquid delivery element and connected with the control device; and / or The second flushing unit comprises: A fourth liquid delivery element connected with the first liquid storage unit, the separation device and the control device; A second flow monitoring element arranged on the fourth liquid delivery element and connected with the control device.

8. The exosome culture separation collection system of claim 4, wherein, The filtration unit comprises: A fifth liquid delivery element connected with the separation device and the control device; A fourth storage element connected with the fifth liquid delivery element, the second liquid storage unit and the third liquid storage unit; An electrostatic adsorption element arranged inside the fourth storage element and connected with the control device; A third filtration element arranged inside the fourth storage element and downstream of the electrostatic adsorption element; and / or The second liquid storage unit comprises: A sixth liquid delivery element connected with the filtration unit and the control device; A fifth storage element connected with the sixth liquid delivery element and the reflux unit; and / or The third liquid storage unit comprises: A seventh liquid delivery element connected with the filtration unit and the control device; A sixth storage element connected with the seventh liquid delivery element; and / or The reflux unit comprises: An eighth liquid delivery element connected with the second liquid storage unit, the culture device and the control device; A third flow monitoring element arranged on the eighth liquid delivery element and connected with the control device.

9. An exosome culture separation collection method applied to the exosome culture separation collection system according to any one of claims 4 to 8, characterized by, The method comprises: Placing a cell culture solution into the culture device, for culturing exosomes; In a case where the culture time of the culture device reaches a preset culture time threshold, the control device controls the flushing device to provide flushing liquid into the interior of the culture device, so that the cell culture solution of the culture device is completely introduced into the separation device, wherein the composition of the flushing liquid is the same as that of the cell culture solution; The control device controls the separation device to perform multi-stage separation on the cell culture solution to obtain the exosomes and the secondary cell culture solution, wherein, in the case that the separation device performs multi-stage separation, the control device controls the flushing device to provide flushing liquid to the inside of the separation device; In the case that the separation device completes multi-stage separation, the control device controls the collection device to collect the exosomes separated by the separation device; In the case that the separation device completes first-stage separation, the control device controls the reflux device to reflux the secondary cell culture solution obtained by the first-stage separation of the separation device to the culture device for the culture device to culture the exosomes.

10. The exosome culture separation collection method of claim 9, wherein, Comprise: Placing the cell culture solution to the culture unit of the culture device for culturing the exosomes; The control device controls the first vibration unit of the culture device to vibrate the culture unit to improve the culture rate; In the case that the culture time of the culture unit reaches the preset culture time threshold, the control device controls the first flushing unit of the flushing device to provide flushing liquid to the inside of the culture unit to make the cell culture solution of the culture unit enter the separation unit of the separation device; The control device controls the vacuum unit, the electrode unit and the second vibration unit of the separation device to perform multi-stage separation on the cell culture solution in the separation unit to obtain the exosomes and the secondary cell culture solution, wherein, in the case that the separation unit performs multi-stage separation, the control device controls the second flushing unit of the flushing device to provide flushing liquid to the separation unit; In the case that the separation unit completes multi-stage separation, the control device controls the first collection unit of the collection device to collect the exosomes of the first-stage separation and the second collection unit of the collection device to collect the exosomes of the multi-stage separation; In the case that the separation unit completes first-stage separation, the control device controls the filtering unit of the reflux device to filter the secondary cell culture solution; The control device controls the second liquid storage unit of the reflux device to collect the secondary cell culture solution filtered by the filtering unit; The control device controls the third liquid storage unit of the reflux device to collect the secondary cell culture solution not filtered by the filtering unit; The control device controls the reflux unit of the reflux device to reflux the secondary cell culture solution collected by the second liquid storage unit to the culture unit.

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