Closed cell separation operation apparatus

Abstract

A closed cell separation operation apparatus, comprising a first pipe body (10) having an air hole and an inlet, a second pipe body (20) having an air hole (231) and an inlet, and a connecting pipe (2

Description

1. Field of the Invention: The present invention relates to centrifugal separation, and more particularly to a closed cell separation operation apparatus with a design that provides a closed, contamination-free environment for both centrifugation and post-centrifugation supernatant separation. The closed cell separation operation apparatus is particularly useful in, for example, cell separation and isolation. 2. Description of the Related Art: Centrifugation has been extensively used in cell-related studies and assays to sort cells in such as bone marrow aspirate concentrate (BMAC), cerebrospinal fluid (CSF), peripheral blood, urine, ascites, cell culture media, etc. As an essential tool for centrifugal separation, a conventional centrifuge tube is made as a capped test tube. Examples may be those traded under the name of Nunc (Thermo Fisher Scientific, the US). In use of such an existing centrifuge tube, a fluid to be subject to centrifugal processing is transferred into it, and then the centrifuge tube is sealed and placed into a centrifuge for processing. After centrifugation, for removing the resulting supernatant (e.g., plasma for whole blood centrifugation) and harvesting the sediment (e.g., blood cells for whole blood centrifugation), or vice versa, an operator has to separate the two parts by unsealing the tube and transferring the supernatant to a spear vessel. However, this operation unavoidably exposes the supernatant and sediment to the ambient environment and brings about risks of contamination. One solution to address the concern is to perform the whole operation in a clean room or a biosafety cabinet (BSC), which is usually costly and relatively immobile, thus unsuitable for clinical use. SUMMARY OF THE INVENTION TECHNICAL PROBLEM Another problem of the conventional practice for post-centrifugation supernatant separation is that spill tends to happen in the process of transferring the content of the centrifuge tube and reduce the volume of the available specimen. This may have adverse effects on the intended study or assay, or in the worst case, makes it impossible to perform the intended study or assay. Hence, there is a need for a novel apparatus that allows post-centrifugation supernatant separation to be performed efficiently and effusively in a contamination-free manner while eliminating the risks of spill and the needs for costly laboratory equipment. SOLUTION TO PROBLEMS TECHNICAL SOLUTION In view of the aforementioned need, the primary objective of the present invention is to provide a closed cell separation operation apparatus that is designed to provide a closed, contamination-free environment for both centrifugation and post-centrifugation supernatant separation while eliminating the risks of spills and the needs for costly laboratory equipment. In order to achieve this and other objectives of the present invention, the disclosed closed cell separation operation apparatus comprises: a first tube body, being a cylinder-like container defined by a peripheral wall, the first tube body having a closed top that sealably communicates with the exterior through a vent and an inlet, wherein the vent is for allowing an air to pass and enter or exit the first tube body and the inlet is for allowing a fluid from an external fluid source to enter the first tube body; and the first tube body having a closed bottom that sealably communicates with the exterior through a centrally formed outlet; a second tube body, being a cylinder-like container defined by a peripheral wall, the second tube body having a closed top that sealably communicates with the exterior through a vent and an inlet, wherein the vent is for allowing an air to pass and enter or exit the second tube body and the inlet is for allowing at least one part of the fluid from the outlet of the first tube body to enter the second tube body and the second tube body having a closed bottom that sealably communicates with the exterior through a centrally formed outlet; and a connecting pipe, being arranged between the outlet of the first tube body and the inlet of the second tube so as to connect the first tube body and a second tube body. Further, the peripheral walls of the first tube body and the second tube body each extend downward beyond the bottom so the peripheral wall and the bottom form a pipe-receiving space around the outlet, and at least one of the connecting pipe and an extension pipe is configured to be at least partially coiled around the outlet of the tube body and received in the pipe-receiving space of tube body. Further, in the closed cell separation operation apparatus, there is further an extension pipe attached to the outlet of the second tube body for connecting the second tube body to a further said first tube body of a further said closed cell separation operation apparatus. Further, in the closed cell separation operation apparatus, at least one of the peripheral walls of the first tube body and the second tube body has a lower edge formed with a notch for the connecting pipe to pass therethrough, so that when the first tube body and the second tube body stands on a plane, an access to the pipe-receiving space of the first tube body and the second tube body is left for the connecting pipe to pass therethrough. Further, in the closed cell separation operation apparatus, at least one of the connecting pipes and the extension pipe is equipped with a valve for closing or opening the pipe, and the valve is a tube clamp, a flow control valve, a multi-way tap connector or an on-off fluid valve. Further, in the closed cell separation operation apparatus, wherein at least one of the bottoms of the first tube body and the second tube body has a downward conical structure with a diameter gradually reduces and reaches a smallest extent thereof at a periphery of the outlet of the first tube body and the second tube body. Further, in the closed cell separation operation apparatus, at least one of the vents of the first tube body and the second tube body is equipped with a filter for filtering the air passing through the vent. Further, in the closed cell separation operation apparatus, there is further an air-pressure source attached to at least one of the vents of the first tube body and the second tube body for pushing air into or drawing air from the first tube body and the second tube body. The air-pressure source is an air pump or an air injector. Further, in the closed cell separation operation apparatus, wherein at least one of the inlets and the vents of the first tube body and the second tube body is configured to be detachably sealed by a seal in an airtight manner. Further, in the closed cell separation operation apparatus, at least one of the inlets and the vents of the first tube body and the second tube body is further provided with a pipe that is equipped with a vale. The valve is a tube clamp, a flow control valve, a multi-way tap connector or an on-off fluid valve. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is perspective view of a closed cell separation operation apparatus according to one embodiment of the present invention; Fig. 2 through Fig. 4 are applied, cross-sectional view of the closed cell separation operation apparatus of the present invention, illustrating the operation of the closed cell separation operation apparatus during centrifugation and post-centrifugation supernatant separation; Fig. 5 is a perspective view of an alternative implementation of the closed cell separation operation apparatus, wherein two closed cell separation operation apparatus of the present invention are connected in series; and Fig. 6 is a perspective view of a closed cell separation operation apparatus according to another embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION While a preferred embodiment provided hereinafter for illustrating the concept of the present invention has been described above, it is to be understood that the components of the embodiment shown in the accompanying drawings are depicted for the sake of easy explanation and need not to be made to exact scale. For the descriptions herein, the singular forms “a”, and “an” include plural referents unless the context clearly indicates otherwise. Referring first to Fig. 1 and Fig. 2, according to one embodiment of the present invention, a closed cell separation operation apparatus primarily comprises a first tube body 10 capped by a first cap 13, a second tube body 20 capped by a second cap 23, and a connecting pipe 24 connecting the two tube bodies 10, 20. As the first tube body 10 and the second tube body 20 are substantively identical in terms of structure, the following description is directed to only one of them for succinctness. The first tube body 10 or the second tube body 20 is a cylinder-like container defined by a peripheral wall 122 or 222. The first tube body 10 or the second tube body 20 has an open top fully open to the exterior through an opening 11 or 21 and a closed bottom 12 or 22 that sealably communicates with the exterior through a centrally formed outlet 121 or 221. Particularly, the bottom 12 or 22 has a funnel-like, or downward conical structure with a diameter that gradually reduces and reaches its smallest extent at the periphery of the outlet 121 or 221. The peripheral wall 122 or 222 of the tube body 10 or 20 extends downward beyond the closed bottom 12 or 22 so that it works with the funnel-like bottom 12 or 22 to form a pipe-receiving space 123 or 223 around the outlet 121 or 221. The lower edge of the peripheral wall 122 or 222 is further formed with a notch 124 or 224, so that when the first tube body 10 or the second tube body 20 stands on a plane, an access to the pipe-receiving space 123 or 223 is created. It is to be noted that the first tube body 10 or the second tube body 20 is sized similar to the existing centrifuge tubes or blood collection tubes, so as to be compatible with existing centrifuges. Similarly, since the first cap 13, a second cap 23 are substantively identical in terms of structure, the following description is directed to only one for succinctness. The first cap 13 or the second cap 23 is configured to cap the first tube body 10 or the second tube body 20 from top and has a vent 131 or 231 and an inlet 132 or 232. As detailed below, each of the vent 131 or 231 and the inlet 132 or 232 is configured to be removably connected to pipes in used and / or detachably capped by a seal 80 in an airtight manner when not in use. It is to be understood that while the present embodiment has the first tube body 10 or the second tube body 20 capped by the cap 13 or 23 formed with the vent 131 or 231 and the inlet 132 or 232, the disclosed closed cell separation operation apparatus in another embodiment may have the vent 131 or 231 and the inlet 132 or 232 directly formed on a closed top surface of the first tube body 10 or the second tube body 20 instead of using the cap 13 or 23 to close the first tube body 10 or the second tube body 20. The connecting pipe 24 is arranged between the outlet 121 of the first tube body 10 and the inlet 232 of the second tube body 20 to connect the first tube body 10 and the second tube body 20 for liquid communication. The connecting pipe 24 can be partially coiled around the outlet 121 of the first tube body 10 to be well received in the pipe-receiving space 123 of the first tube body 10. With the notch 124 formed on the peripheral wall 122 of the first tube body 10, the connecting pipe 24 can extend between the first tube body 10 and the second tube body 20 without preventing the first tube body 10 from standing on any plane stably. The connecting pipe 24 is further equipped with a valve 60 for closing or opening the connecting pipe 24. In use, with the connecting pipe 24 closed by the valve 60, a feeding pipe 40 communicated with a source of a fluid to be processed (not shown), such as a filled blood bag, a blood gas syringe or a specimen tube, is connected to the inlet 132 of the first tube body 10. At this time, the vent 131 of the first tube body 10 remains open, so that the fluid to be processed (e.g., whole blood) can be transferred to the first tube body 10, as shown in Fig. 2. Herein, the vent 131 of the first tube body 10 may be further provide with a filter 31 for preventing foreign objects from entering the of the first tube body 10 and thereby ensuring a contamination-free operation environment. Specifically, as shown in Fig. 2, a venting pipe 30 equipped with a filter 31 is attached to the vent 131 of the first tube body 10. The venting pipe 30 may be also opened or released by a valve 60. After the fluid is filled into the first tube body 10, the inlet 132 of the first tube body 10 can be capped using a seal 80 directly in an airtight manner, or the feeding pipe 40 remained connected with the inlet 132 of the first tube body 10 is capped using a seal 80 in an airtight manner, as shown Fig. 1. Similarly, the vent 131 of the first tube body 10 can be capped using a seal 80 directly in an airtight manner, or the venting pipe 30 remained connected with the vent 131 is capped using a seal 80 in an airtight manner, as shown Fig. 1. It is to be noted that when the connecting pipe 24, the venting pipe 30, and the feeding pipe 40 are closed by their respective valves 60, the interior of the first tube body 10 is sealed from the exterior and forms a closed environment. Also, the second tube body 20 is now with its vent 231 and outlet 221 sealed and isolated from the exterior. Then the entire closed cell separation operation apparatus can be moved into a centrifuge (not shown) for centrifugation. Therein, the first tube body 10 and the second tube body 20 are seated in two adjacent places in the centrifuge, with the connecting pipe 24 partially coiled around the outlet 121 and received in the pipe-receiving space 123 of the first tube body 10. This prevents the connecting pipe 24 from undesired excessive movement during centrifugation. Now referring to Fig. 3, the fluid in the first tube body 10 has separated into a supernatant (e.g., plasma for whole blood centrifugation) and a sediment (e.g., blood cells for whole blood centrifugation). Then, for the sediment to move into the second tube body 20, the valve 60 on the connecting pipe 24 is open and the caps 80 are removed from the venting pipe 30 of the first tube body 10 and the vent 231 of the second tube body 20, respectively. Optionally, an air-pressure source 70 may be removably connected to the vent 231 of the second tube body 20 for accelerated migration of the sediment with a drawing force. Since the bottom 12 of the first tube body 10 has the funnel-like, or downward conical, structure, and the outlet 121 is at the bottom-most position of the bottom 12 of the first tube body 10, the migration of the sediment can be performed precisely without having the supernatant unintentionally move into the second tube body 20. Besides, as the sediment directly flows into the second tube body 20, the risk of spills can be completely eliminated. After the sediment enters the second tube body 20, the connecting pipe 24 is closed by its valve 60 again, and the vent 231 of the second tube body 20 with the air-pressure source 70 having been detached is capped again, so as to seal the supernatant and the sediment in the first tube body 10 and the second tube body 20, respectively. Consequently, all these enable effective and efficient isolation of the sediment form the supernatant, as shown in Fig. 4. Referring to Fig. 5, for various applications where multiple separations are required, another closed cell separation operation apparatus of the present invention may be connected in series to the first closed cell separation operation apparatus. Specifically, an extension pipe 50 may be used to connect the first tube body 10 of the further closed cell separation operation apparatus. In this case, the extension pipe 50 may be coiled around the outlet 221 and received in the pipe-receiving space 223 of the second tube body 20 during centrifugation as described with respect to the connecting pipe 24. Fig. 6 depicts another embodiment of the closed cell separation operation apparatus of the present invention. As shown in Fig. 6, the closed cell separation operation apparatus is further provided with a second venting pipe 90 attached to the vent 231 of the second tube body 20. The second venting pipe 90 is equipped with a further valve 60 for closing or opening it. The second venting pipe 90 is connected to an additional filter 91 for preventing foreign objects from entering the second tube body 20 and further ensuring a contamination-free operation environment. In this case, an air-pressure source 70, as described previously may be attached to the vent 131 of the first tube body 10 for pushing the content in the first tube body 10 into the second tube body 20. The foregoing valves 60 may each independently be a tube clamp, a flow control valve, a multi-way connector or an on-off fluid valve, without limitation. The air-pressure source 70 attached to the vent 131 of the first tube body 10 and / or the vent 231 of the second tube body 20 may be an air pump or an air injector or any other device that can change the pressure in the first tube body 10 and the second tube body 20 to facilitate fluid flow from the first tube body 10 to the second tube body 20. At last, centrifugation is widely used in various medical and biological applications and well known to those skilled in the art, so the related operation and principle are not described in detail for the sake of succinctness. Likewise, the cell isolation technology involving in the use of the disclosed closed cell separation operation apparatus is also known in the art and not discussed herein. With the configurations described above, the closed cell separation operation apparatus achieves the objectives of the present invention by providing a closed, contamination-free environment for both centrifugation and post-centrifugation supernatant separation while eliminating the risks of spills and the needs for costly laboratory equipment. The present invention has been described regarding the preferred embodiments and it is understood that the embodiments are not intended to limit the scope of the present invention. Moreover, all modifications, equivalent changes or improvements which do not depart from the concept of the present invention should be encompassed by the appended claims.

Claims

1. A closed cell separation operation apparatus, comprising:first tube body, being a cylinder-like container defined by a peripheral wall, the first tube body having a closed top that sealably communicates with the exterior through a vent and 5 an inlet, wherein the vent is for allowing air to pass and enter or exit the first tube body and the inlet is for allowing fluid from an external fluid source to enter the first tube body; and the first tube body having a closed bottom that sealably communicates with the exterior through a central outlet;10 a second tube body, being a cylinder-like container defined by a peripheral wall, the second tube body having a closed top that sealably communicates with the exterior through a vent and an inlet, wherein the vent is for allowing an air to pass and enter or exit the second tube body and the inlet is for15 allowing at least one part of the fluid from the outlet of the first tube body to enter the second tube body and the second tube body having a closed bottom that sealably communicates with the exterior through a centrally formed outlet; anda connecting pipe, being arranged between the outlet of the first tube body and the inlet of the second tube body, so as to link the first tube body (10) and the second tube body.

2. The closed cell separation operation apparatus of claim 1, wherein the peripheral wall of the first tube body extends downward the bottom of the first tube body so the peripheral wall and the bottom form a pipe-receiving space around the outlet of the first tube body.

3. The closed cell separation operation apparatus of claim 2, wherein the peripheral wall of the first tube body has a lower edge formed with a notch for the connecting pipe to pass therethrough.

4. The closed cell separation operation apparatus of claim 1, further comprising an extension pipe attached to the outlet of the second tube body for connecting the second tube body to a further said first tube body of a further said closed cell separation operation apparatus.

5. The closed cell separation operation apparatus of claim 4, wherein the peripheral wall of the second tube body extends downward the bottom of the second tube body so theperipheral wall and the bottom form a pipe-receiving space around the outlet of the second tube body.

6. The closed cell separation operation apparatus of claim 5, wherein the peripheral wall of the second tube body has a lower edge formed with a notch for the connecting pipe to pass therethrough.

7. The closed cell separation operation apparatus of claim 1, wherein the connecting pipe is equipped with a valve for closing or opening the connecting pipe.

8. The closed cell separation operation apparatus of claim 4, wherein the extension pipe is equipped with a valve for closing or opening the extension pipe.

9. The closed cell separation operation apparatus of claim 2, wherein the bottom of the first tube body has a downward conical structure with a diameter gradually reduces and reaches a smallest extent thereof at a periphery of the outlet of the first tube body.

10. The closed cell separation operation apparatus of claim 5, wherein the bottom of the second tube body has a downward conical structure with a diameter gradually reducesand reaches a smallest extent thereof at a periphery of the outlet of the second tube body11. The closed cell separation operation apparatus of claim 1, wherein at least one of the vents of the first tube body (and the second tube body is equipped with a filter for filtering the air passing through the vent.

12. The closed cell separation operation apparatus of claim 1, wherein at least one of the vents of the first tube body and the second tube body is equipped with an air-pressure source for pushing an air into or drawing an air from the first tube body and the second tube body.

13. The closed cell separation operation apparatus of claim 1, wherein at least one of the inlets and the vents of the first tube body and the second tube body is configured to be detachably sealed by a seal in an airtight manner.

14. The closed cell separation operation apparatus of claim 1, wherein at least one of the inlets and the vents of the first tube body and the second tube body is further provided with a pipe that is equipped with a vale.PCT / CN 2022 / 082170A. CLASSIFICATION OF SUBJECT MATTER C12M 3 / 08(2006.01)1 According to International Patent Classification (IPC) or to both national classification and IPC B. FIELDS SEARCHED Minimum documentation searched (classification system followed by classification symbols) C12M Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched Electronic data base consulted during the international search (name of data base and, where practicable, search terms used) CNTXT; CNABS; ENTXT: ENTXTC; VEN: >lW, « IX, A®, Affifi, W, H h®®, centrifugal, two tube?, supemat+, second tube, blood, separat+, tube, blood, cell?, double tube?, in series C. DOCUMENTS CONSIDERED TO BE RELEVANT Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. Y KR 1020140131211 A (DONGGUK UNIVERSITY INDUSTRY-ACADEMIC COOPERATION FOUNDATION et al.) 12 November 2014 (2014-11-12) description, paragraphs 1, 7, 31, 37 and 49, and figures 1-7 1-14 Y KR 101091052 Bl (COLLEGE OF MEDICINE POCHON CHA UNIVERSITY INDUSTRYACADEMIC COOPERATION FOUNDATION) 08 December 2011 (2011-12-08) embodiment 2, and figure 3 1-14 Y KR 1020110111059 A (CHABIOMED CO., LTD.) figure 6 10 October 2011 (2011-10-10) 1-14 Y CN 213943196 U (SUZHOU DAISHI ELECTRONIC INFORMATION TECHNOLOGY CO., LTD.) 13 August 2021 (2021-08-13) embodiment 1, figure 1 1-14 Y CN 212791027 U (WEST CHINA HOSPITAL, SICHUAN UNIVERSITY) 26 March 2021 (2021-03-26) claims 1-10, and figures 1-2 1-14 E TW M627564 U (DUOGENIC STEMCELLS CORP.) 01 June 2022 (2022-06-01) description, paragraphs 12-27, and figures 1-6 1-14 | | Further documents are listed in the continuation of Box C. | J | See patent family annex. * Special categories of cited documents: “A” document defining the general state of the art which is not considered to be of particular relevance “E” earlier application or patent but published on or after the international filing date “L” document which may throw doubts on priority claim(s) or which is cited to establish the publication date of another citation or other special reason (as specified) “O” document referring to an oral disclosure, use, exhibition or other means “P” document published prior to the international filing date but later than the priority date claimed “T” later document published after the international filing date or priority date and not in conflict with the application but cited to understand the principle or theory underlying the invention “X” document of particular relevance; the claimed invention cannot be considered novel or cannot be considered to involve an inventive step when the document is taken alone “Y” document of particular relevance; the claimed invention cannot be considered to involve an inventive step when the document is combined with one or more other such documents, such combination being obvious to a person skilled in the ait document member of the same patent family Date of the actual completion of the international search 06 December 2022 Date of mailing of the international search report 26 December 2022 Name and mailing address of the ISA / CN China National Intellectual Property Administration (ISA / CN) No. 6, Xitucheng Road, Jimenqiao, Haidian District, Beijing 100088, China Facsimile No. (86-10)62019451 Authorized officer Telephone No.PCT / CN2022 / 082170C. DOCUMENTS CONSIDERED TO BE RELEVANTCategory* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. A CN 211358875 U (GUO YANQING) 28 August 2020 (2020-08-28) entire document 1-14 A CN 106929399 A (EASTERN UNION STEM CELL &GENE ENGINEERING CO., LTD.) 07 July 2017 (2017-07-07) entire document 1-14 A CN 205659694 U (HANGZHOU SHANHATTANG HEALTH TECHNOLOGY CO., LTD.) 26 October 2016 (2016-10-26) entire document 1-14 A CN 212159203 U (SYNERMORE BIOLOGICS (SUZHOU) CO., LTD.) 15 December 2020 (2020-12-15) entire document 1-14INTERNATIONAL SEARCH REPORT Information on patent family membersInternational application No.PCT / CN2022 / 082170Patent document cited in search report Publication date (day / month / year) Patent family member)s) Publication date (day / month / year) KR 1020140131211 A 12 November 2014 None KR 101091052 Bl 08 December 2011 KR 1020100095952 A 01 September 2010 KR 1020110111059 A 10 October 2011 WO 2011122844 A2 06 October 2011 CN 213943196 U 13 August 2021 None CN 212791027 U 26 March 2021 None TW M627564 U 01 June 2022 None CN 211358875 U 28 August 2020 None CN 106929399 A 07 July 2017 None CN 205659694 U 26 October 2016 None CN 212159203 U 15 December 2020 None

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