CN209048767U - A kind of stem cell in vitro enrichment isolation system - Google Patents

A kind of stem cell in vitro enrichment isolation system Download PDF

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Publication number
CN209048767U
CN209048767U CN201821486972.6U CN201821486972U CN209048767U CN 209048767 U CN209048767 U CN 209048767U CN 201821486972 U CN201821486972 U CN 201821486972U CN 209048767 U CN209048767 U CN 209048767U
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China
Prior art keywords
stem cell
filter cartridge
isolation system
shell
enrichment isolation
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Expired - Fee Related
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CN201821486972.6U
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Chinese (zh)
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干耀恺
戴尅戎
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Ninth Peoples Hospital Shanghai Jiaotong University School of Medicine
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Ninth Peoples Hospital Shanghai Jiaotong University School of Medicine
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Abstract

The utility model discloses a kind of stem cell in vitro enrichment isolation system, the filter cartridge group being connected into including several filter cartridges and sealing for holding the shell of filter cartridge group, the adsorbed film for adsorbing stem cell is equipped in each filter cartridge;The peristaltic pump that filter cartridge group can be located at outside shell with one forms closed loop circulating system;Stem cell in vitro enrichment isolation system further include: for holding filter cartridge, stem cell is set to shed into the oscillator of stem cell suspension from adsorbed film, and it is a set of keep filter cartridge in parallel, for stem cell to be eluted from adsorbed film into stem cell suspension and recycles the elution pipeline of stem cell suspension.Its technical effect is that: its efficiency that can effectively improve stem cell in vitro enrichment isolation quickly sticks technology by stem cell and quickly collects the suspension that acquisition is enriched with stem cell, meets the case where clinic only needs stem cell suspension requirement.

Description

A kind of stem cell in vitro enrichment isolation system
Technical field
The utility model relates to a kind of stem cell in vitro enrichment isolation systems in stem-cell therapy field.
Background technique
The autologous stem cells beneficiation technologies of Recent study discovery, i.e., without the stem cell of in vitro culture, direct Hui Zhi, Not only it is avoided that immunological rejection, and the autologous stem cells being enriched with can use local microenvironment and realize its directional induction Thus box regeneration brings hope to the treatment of some diseases.Its biggest advantage is that being not necessarily in vitro culture, can perform the operation The technical issues of same period carries out, and avoids complex operations and relevant ethnics Problem, this is that conventional method can not substitute and compare Quasi-.With deepening continuously for stem-cell research, with stem cell (bone marrow stem cells, BMSCs) conduct The cell therapy of seed cell receives more and more attention.BMSCs promotes the zoopery of Bone Defect Repari more to be affirmed Effect, but carry out clinical treatment be still subject to many limitations.First, when being treated with autogenous bone marrow blood merely, because of unit bodies BMSCs amount reduces curative effect less in product marrow blood.Second, it is expanded according to cell injuring model, then patient will wait one section Time just can be carried out back plant operation, generally about need for 3 weeks, the needs for not being able to satisfy clinical emergency or treating in time;And body Outer culture must avoid the pollution of various microorganisms, to the more demanding of environment and equipment, therefore increase medical expense;In addition For in vitro culture often using Cow placeta serum as culture medium, this also brings the ethics dispute of clinical cytology treatment;Without training in vitro The feeding BMSCs stem cell enrichment screening technique that high concentration can be obtained in art is developed.
Cell enrichment screening technique mainly has at present: flow cytometric sorting method, magnetic activated cell seperation, adherent screening method And density-gradient centrifugation method.Flow cytometric sorting method is the antigen marker knot using special fluorescence antibody and cell surface It closes, then stem cell is sorted using flow cytometer.Although it separates cell rapidly, purity is high, somewhat expensive, The trace cell too low to content is difficult to separate, and cannot handle a large amount of cell sample, to obtain sterile stem cell suspension It is highly difficult.
Magnetic activated cell seperation is using micro- magnetic bead not only in combination with protein, but also the characteristics of can be attracted by magnet, by antibody It is coated on magnetic bead, by forming antigen-antibody magnetic bead immune complex in conjunction with stem cell surface specific antigen.When this It when compound passes through splitter, under magneticaction, i.e., separates with mother liquor, so that stem cell be made to separate with other substances, reaches The purpose of screening.It is larger to the activity influence of cell, and the rate of recovery only has 60%-75%, may result in cell complete deactivation, Equally operate it is more complex, it is expensive.
Adherent screening method is the most common bone marrow stroma stem cell screening technique in current laboratory, by training marrow blood It supports and is cultivated in ware, bone marrow stroma stem cell is adherent after 24-48h, whereby can be by bone marrow stroma stem cell and other not adherent bones Myelocyte separation.It needs the time longer, is related to the in vitro culture of cell, and clinical use acquires a certain degree of difficulty.
Density-gradient centrifugation method is the method for common concentration and separation cell, utilizes the difference of different cell component density gradients It is different, cell can be made in different levels concentration by centrifugation, the separating liquid of different specific weight be used in combination, by the cell of different specific weight Carry out multi_layer extraction.It needs finally to need to wash away separating medium again using separating medium, to keep the sterile need of whole operation Want special equipment, somewhat expensive.
Utility model content
The purpose of this utility model is to overcome the deficiencies in the prior art, provide a kind of stem cell in vitro enrichment isolation system System, can directly obtain stem cell suspension, more be able to satisfy clinical a variety of needs, due to ceramic without using biological stephanoporate, The efficiency of stem cell enrichment screening can be effectively improved, stem cell enrichment screening is reduced, obtains cost and the time of stem cell suspension, Meet the requirement of emergency treatment clinical operation.
Realizing a kind of technical solution of above-mentioned purpose is: a kind of stem cell in vitro enrichment isolation system, including several filterings The shell of box and a sealing;
The filter cartridge can be sequentially connected in series from bottom to up as a filter cartridge group in the shell, and the filter cartridge group can The peristaltic pump being located at outside the shell in the shell with one forms closed loop circulating system;
The adsorbed film for adsorbing stem cell is equipped in each filter cartridge;
The stem cell in vitro enrichment isolation system further include:
For holding the filter cartridge, stem cell is made to shed into stem cell suspension from the adsorbed film of the filter cartridge Oscillator and it is a set of keep the filter cartridge in parallel, for eluting stem cell into stem cell suspension, and by stem cell suspension The elution pipeline recycled out of described filter cartridge.
Further, the upper and lower ends of the filter cartridge are correspondingly provided with closed upper sealed interface and lower sealed interface.
Further, it is connected between two filter cartridges of arbitrary neighborhood in the filter cartridge group by jointing, institute State the lower sealed interface of the filter cartridge of the top connection top of jointing, the filtering of the bottom end connection lower section of the jointing The upper sealed interface of box.
Further, the elution pipeline includes the upper elution pipe that can be connect with the upper sealed interface of the filter cartridge Road, and the lower elution pipeline that can be connect with the lower sealed interface of the filter cartridge.
Further, the filter cartridge is that top surface area is big, the small conical filter box of base area.
Further, the top of the shell is equipped with top interface, filter cartridge group described in the top orifice, and leads to Cross the import of peristaltic pump described in bleeding pipeline connection;
The bottom end of the shell is equipped with bottom interface, and the bottom interface is equipped with triple valve, and the triple valve is connected to institute Filter cartridge group is stated, and passes through the outlet of peristaltic pump described in blood back pipeline connection.
Further, the adsorbed film has the double-layer structure being made of two layers of perforated membrane, and the aperture of every layer of perforated membrane is equal It is 5~8 times of stem cell diameter.
Using a kind of technical solution of stem cell in vitro enrichment isolation system of the utility model, including several filterings Cassette in series at filter cartridge group and sealing for holding the shell of filter cartridge group, be equipped in each filter cartridge for inhaling The adsorbed film of attached stem cell;The peristaltic pump that filter cartridge group can be located at outside shell with one forms closed loop circulating system;It is dry thin The outer enrichment isolation system of cell space further include: for holding filter cartridge, stem cell is made to shed into stem cell suspension from adsorbed film Oscillator and it is a set of keep filter cartridge in parallel, for eluting into stem cell suspension and recycling stem cell from adsorbed film The elution pipeline of stem cell suspension.Its technical effect is that: its efficiency that can effectively improve stem cell in vitro enrichment isolation, by dry Cell, which quickly sticks technology, quickly to be collected to obtain and is enriched with the suspension of stem cell, rather than stem cell and material is compound Object, and the cost of stem cell in vitro enrichment isolation is reduced, meet the case where clinic only needs stem cell suspension requirement.
Detailed description of the invention
Fig. 1 is a kind of schematic diagram of the closed loop circulating system of stem cell in vitro enrichment isolation system of the utility model.
Fig. 2 is a kind of schematic diagram of the oscillating separator of stem cell in vitro enrichment isolation system of the utility model.
Fig. 3 is a kind of schematic diagram of the elution pipeline of stem cell in vitro enrichment isolation system of the utility model.
Specific embodiment
Please refer to Fig.1 to Fig.3, the inventor of the utility model in order to preferably to the technical solution of the utility model into Row understands, below by specifically embodiment, and will be described in detail with reference to the accompanying drawings:
Referring to Fig. 1, a kind of stem cell enrichment screening system of the utility model includes, several filter cartridges 2, one are close The shell 1 of envelope and the peristaltic pump 3 outside shell 1.The quantity of filter cartridge 2 is usually three to six.The upper end of filter cartridge 2 is set There is closed upper sealed interface 21, the lower end of filter cartridge 2 is equipped with closed lower sealed interface 22, the filter cartridge 2 of top It can be connected by medical jointing 23 between lower sealed interface 22 and the upper sealed interface 21 of the filter cartridge 2 of lower section.It is several A filter cartridge 2 can be sequentially connected in series from bottom to up in shell 1, form a filter cartridge group.It is equipped with and is used in each filter cartridge 2 Adsorb the adsorbed film 20 of stem cell.
The top of shell 1 is closed by lid 11, and the center of lid 11 is equipped with top interface 12, and the bottom of shell 1 is equipped with Bottom interface 13.The filter cartridge group is placed in shell 1, the upper sealed interface of the filter cartridge 2 of top in the filter cartridge group 21 are located at the bleeding pipeline 31 outside shell 1 by the connection of top interface 12, and bleeding pipeline 31 is connected to the import of peristaltic pump 3.Filtering Triple valve 14 of the lower sealed interface 22 of the filter cartridge 2 of lowermost end by the connection of bottom interface 13 outside shell 1 in box group, three Port valve 14 is connected to blood back pipeline 32, and blood back pipeline 32 is connected to the outlet of peristaltic pump 3.Filter cartridge group forms closed loop with peristaltic pump 3 and follows Loop system, after marrow blood recycles 10 minutes in the closed loop circulating system, in the filter cartridge group in each filter cartridge 2 Adsorbed film 20 can at least adsorb in marrow blood, 85% stem cell completes the acquisition of stem cell.Meanwhile Triple valve 14 is also connected into blood vessel road 33, and marrow blood is by entering the closed loop circulating system into blood vessel road 33, and from institute State closed loop circulating system recycling marrow blood.
Adsorbed film 20 has the double-layer structure being made of two layers of perforated membrane, and the aperture of every layer of perforated membrane is stem cell diameter 5~8 times.20 material of adsorbed film is selected from bata-tricalcium phosphate, natural non-organic bone NNB, decalcified bone matrix DBM, forging bone, poly- cream Acid-polyglycolic acid, collagen or freeze-drying cancellous bone.Above-mentioned material has good characterization of adsorption to stem cell.
The closed loop circulating system is using adsorbed film 20 for the characteristic of stem cell quick adsorption, therefore marrow blood During circulation in the closed loop circulating system, the cell in marrow blood can be adsorbed 20 quick adsorption of film, improve The survival rate of stem cell, improves the acquisition of stem cell in the efficiency and collection process of stem cell enrichment isolation Amount, reduces the blood sampling volume of patient, ensures the safety of patient, mitigate the pain and risk of patient, reduces stem cell enrichment screening Cost, while reducing in collection process a possibility that stem cell pollutes.
A kind of stem cell enrichment screening system of the utility model includes oscillator 4, by marrow blood cyclic process In, by the absorption of adsorbed film 20, after having acquired stem cell, filter cartridge group is taken out out of shell 1, and medical connection connects First 23 separate with corresponding filter cartridge 2, and filter cartridge group is disassembled as single filter cartridge 2, from upper sealed interface 21 to corresponding The physiological saline of 3~5ml is added in filter cartridge 2,0.01wt.% anticoagulant factor or 0.01wt.% liver can be contained in physiological saline Other anti-coagulants of element or 0.01wt.% prevent stem cell outstanding in stem cell to guarantee the survival rate of stem cell Assemble in liquid.Then filter cartridge 2 puts into oscillator 4 makes stem cell fall off from adsorbed film 20 by the oscillation of oscillator 4, Into in physiological saline, stem cell suspension is formed.In order to improve the efficiency of stem cell enrichment screening, oscillator 4, which is selected, once may be used To complete the isolated centrifugation oscillator of stem cell and adsorbed film 20 in multiple filter cartridges 2, therefore the quantity of filter cartridge 2 is excellent Even number is selected, and filter cartridge 2 is that top surface area is big, the small conical filter box of base area.
A kind of stem cell enrichment screening system of the utility model further includes a set of elution pipeline for making the parallel connection of filter cartridge 2 5.Elute pipeline 5 include the upper elution pipeline 51 being connect with the upper sealed interface 21 of each filter cartridge 2, and with each filter cartridge The lower elution pipeline 52 that 2 lower sealed interface 22 connects.It is upper elution pipeline 51 and each filter cartridge 2 upper sealed interface 21 it Between, shut-off valve is equipped between lower elution pipeline 52 and the lower sealed interface 22 of each filter cartridge 2.Upper elution pipeline 51 be used for The physiological saline of 1~2ml of injection in each filter cartridge 2, in physiological saline can containing 0.01wt.% anticoagulant factor or Other anti-coagulants of 0.01wt.% heparin or 0.01wt.% make to remain for eluting the adsorbed film 20 in filter cartridge 2 Remaining stem cell is separated with adsorbed film 20, into stem cell suspension.Stem cell suspension can be from lower elution pipeline 52 Recycling is used for subsequent treatment.
For the utility model a kind of stem cell enrichment screening system operation under aseptic conditions into can be effective The efficiency for improving stem cell enrichment screening, meets requirement used as needed in clinical operation.
Those of ordinary skill in the art it should be appreciated that more than embodiment be intended merely to illustrate that this is practical new Type, and be not used as the restriction to the utility model, as long as in the spirit of the utility model, to the above Variation, the modification of embodiment will all be fallen in the Claims scope of the utility model.

Claims (7)

1. a kind of stem cell in vitro enrichment isolation system, including several filter cartridges and a shell sealed, it is characterised in that:
The filter cartridge can be sequentially connected in series from bottom to up as a filter cartridge group in the shell, and the filter cartridge group can be in institute It states the peristaltic pump being located at outside the shell in shell with one and forms closed loop circulating system;
The adsorbed film for adsorbing stem cell is equipped in each filter cartridge;
The stem cell in vitro enrichment isolation system further include:
For holding the filter cartridge, stem cell is made to shed into the oscillation of stem cell suspension from the adsorbed film of the filter cartridge Device and it is a set of keep the filter cartridge in parallel, for eluting stem cell into stem cell suspension, and by stem cell suspension from institute State the elution pipeline recycled in filter cartridge.
2. a kind of stem cell in vitro enrichment isolation system according to claim 1, it is characterised in that: the filter cartridge it is upper Lower both ends are correspondingly provided with closed upper sealed interface and lower sealed interface.
3. a kind of stem cell in vitro enrichment isolation system according to claim 2, it is characterised in that: in the filter cartridge group Two filter cartridges of arbitrary neighborhood between connected by jointing, the filter cartridge of the top of jointing connection top Lower sealed interface, the upper sealed interface of the filter cartridge of the bottom end connection lower section of the jointing.
4. a kind of stem cell in vitro enrichment isolation system according to claim 2, it is characterised in that: the elution pipeline packet The upper elution pipeline that can be connect with the upper sealed interface of the filter cartridge is included, and can be connected with the lower sealed interface of the filter cartridge The lower elution pipeline connect.
5. a kind of stem cell in vitro enrichment isolation system according to claim 1, it is characterised in that: the filter cartridge is top Face area is big, the small conical filter box of base area.
6. a kind of stem cell in vitro enrichment isolation system according to claim 1, it is characterised in that: the top of the shell Equipped with top interface, filter cartridge group described in the top orifice, and pass through the import of peristaltic pump described in bleeding pipeline connection;
The bottom end of the shell is equipped with bottom interface, and the bottom interface is equipped with triple valve, and the triple valve is connected to the mistake Box group is filtered, and passes through the outlet of peristaltic pump described in blood back pipeline connection.
7. a kind of stem cell in vitro enrichment isolation system according to claim 1, it is characterised in that: the adsorbed film has The double-layer structure being made of two layers of perforated membrane, the aperture of every layer of perforated membrane are 5~8 times of stem cell diameter.
CN201821486972.6U 2018-09-12 2018-09-12 A kind of stem cell in vitro enrichment isolation system Expired - Fee Related CN209048767U (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108785779A (en) * 2018-09-12 2018-11-13 上海交通大学医学院附属第九人民医院 A kind of stem cell in vitro enrichment isolation system and method

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108785779A (en) * 2018-09-12 2018-11-13 上海交通大学医学院附属第九人民医院 A kind of stem cell in vitro enrichment isolation system and method

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