CN205473829U - Interact's culture apparatus between cell culture and cell - Google Patents
Interact's culture apparatus between cell culture and cell Download PDFInfo
- Publication number
- CN205473829U CN205473829U CN201620298442.3U CN201620298442U CN205473829U CN 205473829 U CN205473829 U CN 205473829U CN 201620298442 U CN201620298442 U CN 201620298442U CN 205473829 U CN205473829 U CN 205473829U
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- Prior art keywords
- cell
- culture
- detachable
- shaping piece
- semipermeable membrane
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 238000004113 cell culture Methods 0.000 title claims abstract description 32
- 238000003501 co-culture Methods 0.000 claims abstract description 36
- 238000007493 shaping process Methods 0.000 claims description 66
- 239000012528 membrane Substances 0.000 claims description 44
- 230000008611 intercellular interaction Effects 0.000 claims description 21
- 238000003780 insertion Methods 0.000 claims description 5
- 230000037431 insertion Effects 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 4
- 238000007789 sealing Methods 0.000 claims description 4
- 238000005192 partition Methods 0.000 claims description 3
- 239000004417 polycarbonate Substances 0.000 claims description 2
- 229920000515 polycarbonate Polymers 0.000 claims description 2
- 239000011148 porous material Substances 0.000 claims description 2
- 238000002955 isolation Methods 0.000 claims 2
- 238000000465 moulding Methods 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 52
- 210000004881 tumor cell Anatomy 0.000 abstract description 27
- 238000000034 method Methods 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 8
- 230000001464 adherent effect Effects 0.000 abstract description 4
- 238000013508 migration Methods 0.000 abstract description 4
- 230000005012 migration Effects 0.000 abstract description 4
- 230000001954 sterilising effect Effects 0.000 abstract description 3
- 125000006850 spacer group Chemical group 0.000 abstract 1
- 238000004659 sterilization and disinfection Methods 0.000 abstract 1
- 206010028980 Neoplasm Diseases 0.000 description 14
- 230000006872 improvement Effects 0.000 description 9
- 206010006187 Breast cancer Diseases 0.000 description 4
- 208000026310 Breast neoplasm Diseases 0.000 description 4
- KDXKERNSBIXSRK-RXMQYKEDSA-N D-lysine Chemical compound NCCCC[C@@H](N)C(O)=O KDXKERNSBIXSRK-RXMQYKEDSA-N 0.000 description 4
- 230000005484 gravity Effects 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 108010082117 matrigel Proteins 0.000 description 4
- 230000007547 defect Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 201000008275 breast carcinoma Diseases 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 210000001711 oxyntic cell Anatomy 0.000 description 2
- 239000002985 plastic film Substances 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 241001149900 Fusconaia subrotunda Species 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 150000004651 carbonic acid esters Chemical class 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 210000003322 glomus cell Anatomy 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000016507 interphase Effects 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 210000005075 mammary gland Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 230000005748 tumor development Effects 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The utility model discloses an interact's culture apparatus between cell culture and cell, the device be including cultivateing the board, train and to be provided with a culture hole at least on the board, at least one is cut apart into culture hole the separated pellicle spacing block of a plurality of cell culture room and lies in the separated spacing block that can separate pellicle spacer both sides. The device's application method is that the sterilization back is inserted at culture hole and can be separated the pellicle spacing block, can separate the spacing block, cultivate tumor cells a cell culture room of cutting apart, afterwards at another cell culture room adherent cell, separate and to separate the spacing block, make tumor cells and adherent cell co -culture, continuously co -culture, observe the tumor cells balling -up condition to and the tumor cells of balling -up from the migration condition of a cell culture room to another cell culture room. Utilize this culture apparatus to carry out interact effect observation between tumor cells balling -up cultivation and cell, easy operation is practical, effect clear.
Description
Technical field
This utility model relates to biological technical field, relates in particular to a kind of cell and cultivates and thin
The co-culture device that intercellular interacts.
Background technology
The cultivation of ordinary two dimensional cell is utilized to study tumor development machine-processed and the most internal
Environment is very different, as cell divides with substrate interphase interaction, cell with cell, cell
Change.Animal model, owing to there is species variation with human body, can not well repeat human body special
Levy, such as human tumor growth's transfer, Drug therapy reaction, immunoreation and tumor stem cell
(TSCs) differentiation etc..
Three-dimensional cell culture technology is simple to operation, it is possible to simulated in vivo environment feature.In three-dimensional
In cultivation, tumor cell or the TSCs of single cell type can be comprised, it is also possible to comprise many cells
Being such as tumor cell, substrate and immunocyte, such model is cultivated than common two dimension and more can
Simulation tumor true growing state in vivo.In the incubation of cell, dimensional culture can
To promote the connection between cell and iuntercellular and cell and extracellular matrix, Three-dimensional cell culture is than patch
Parietal cell is cultivated and is more easy to be supplied to one, cell close to internal environment, therefore, and dimensional culture skill
Art application in Cultural Course of Tumor Cells receives people and more and more payes attention to, but three
It is a kind of new technique that dimension cell is cultivated, and also has some incomplete place, as lacked at present and it
The three-dimensional nodule cell culture studies of his cell-cell interaction.
In dimensional culture technology, cell balling-up technology is more and more paid attention to.Tumor is thin
Born of the same parents' ball sample aggregation is cultivated, and has another name called multicellular spheriods and cultivates, and referred to as balling-up is cultivated, its purpose
It is that scattered oncocyte is carried out In vitro culture so that it is re-form and be morphologically similar to internal life
Long solid tumor sample tuberosity, does some tumor experiments with this tuberosity (spheroid) and studies closer to body
Interior growth.Dittmer etc. find breast cancer cell (MCF-7) energy in dimensional culture model
Enough attractive interstital stem cells (hMSCs) also strengthen they transfer abilities, and hMSCs also can
The mammary gland ball entering breast carcinoma formation affects growth development and the immunoreation of the latter.But its
The simplest breast cancer cell that hMSCs is added balling-up of experiment observes connecing of the two
Touching reaction, owing to two kinds of cells have been thoroughly mixed in together, this is difficult to follow-up be kept completely separate two
Plant cell, observe the characteristic of two kinds of cells respectively, and cannot accomplish that hMSCs is to breast carcinoma
The observation of cell balling-up capacity.The limitation of this research is cultivated and it with lacking cell balling-up
He has some relations by the co-culture device of cell-cell interaction.Study more lymphocyte at present
Similar Problems is there is also with tumor cell Co-culture.
Traditional Transwell cell is utilized to realize being total to of tumor cell balling-up and other cell
Cultivation there is problems in that the particular/special requirement that (1) cultivates due to tumour 3 D balling-up, and tumor is thin
Born of the same parents can only be confined to the lower floor of Transwell cell, therefore limits other cell to balling-up
The research of some function effects of tumor cell；(2) Transwell cell is the training of levels cell
Support structure, it is impossible to get rid of the defect of gravity interference.
Utility model content
The purpose of this utility model is the defect for above-mentioned prior art, it is provided that a kind of cell
Cultivate and the co-culture device of cell-cell interaction, simple in construction, with low cost, the easiest
Operation, overcomes above-mentioned conventional vertical co-culture device cannot get rid of the balling-up tumor bigger to volume
The defect of cell gravity interference, and solve in prior art tumor cell balling-up and cultivate and be difficult to and it
The problem of its co-culture of cells, it is simple to observe tumor and become glomus cell and other intercellular phase interaction
With.
For achieving the above object, technical solution adopted in the utility model is:
A kind of cell is cultivated and the co-culture device of cell-cell interaction, including culture plate, institute
State on culture plate at least provided with a culture hole, at least one culture hole is divided into several
The detachable semipermeable membrane shaping piece of cell culture chamber and be positioned at detachable semipermeable membrane partition both sides
Detachable shaping piece.
As the improvement to technique scheme, the most parallel on the hole wall of described culture hole set
It is equipped with and sets detachable semipermeable membrane shaping piece, the draw-in groove of detachable shaping piece, described draw-in groove for card
Longitudinally opposed distribution；The height of described draw-in groove is identical with the degree of depth of culture hole, and described detachable half
In the insertion draw-in groove that permeable membrane shaping piece, detachable shaping piece are detachable.
As the improvement to technique scheme, described culture hole and detachable semipermeable membrane shaping piece,
The junction of detachable semipermeable membrane shaping piece, draw-in groove and detachable semipermeable membrane shaping piece, detachable
The junction of semipermeable membrane shaping piece is provided with prevent liquid between cell culture chamber from flowing mutually close
Seal apparatus.This sealing device be can fill in draw-in groove with detachable shaping piece or with detachable half
Permeable membrane shaping piece forms diaphragm seal, the sealing gasket etc. sealed.
As the improvement to technique scheme, described detachable shaping piece is symmetrically distributed in removable
From the both sides of semipermeable membrane shaping piece, the most adjacent with detachable semipermeable membrane shaping piece, at a distance of width
≤0.5mm。
As the improvement to technique scheme, described detachable semipermeable membrane shaping piece is detachable
Be suitable to culture medium or semipermeable membrane shaping piece that cell passes.
As the improvement to technique scheme, described detachable semipermeable membrane shaping piece is poly-carbonic acid
Ester film shaping piece.
As the improvement to technique scheme, the aperture of described detachable semipermeable membrane shaping piece is big
Little for 3-8 μm.
As the improvement to technique scheme, described detachable shaping piece is dismountable without thin
The plastic sheet of born of the same parents' adhesion, height is identical with the culture hole degree of depth.
As the improvement to technique scheme, described culture plate is that 6 culture hole or 12 are cultivated
Hole or the Tissue Culture Plate of 24 culture hole or Tissue Culture Dish.
This cell cultivates the using method of the co-culture device with cell-cell interaction:
S1, co-culture device is carried out autoclaving and ultra-vioket radiation sterilizing；
S2, insert detachable semipermeable membrane shaping piece, detachable shaping piece；
Under S3, sterile working, tumor cell suspension is added the cell cultivation being coated matrigel
Room, makes tumor cell be sunken to matrigel, stablizes grown cultures 3 days in cell culture chamber；
S4, the 4th day time, be coated another cell culture chamber with poly-D-lysine, another kind of patch
Parietal cell is planted in the cell culture chamber being coated poly-D-lysine.37 DEG C of incubated overnight, are allowed to
Adherent growth in cell culture chamber；
S5, separate detachable shaping piece, make tumor cell and attached cell co-culture；
S6, persistently co-culture, observe tumor cell balling-up situation, and the tumor cell of balling-up
From a cell culture chamber to the migration situation of another cell culture chamber.
Compared with prior art, this utility model has the advantage that with good effect and is:
Cell of the present utility model is cultivated and the co-culture device of cell-cell interaction, and 1, knot
Structure is simple and practical, it is to avoid conventional tumor cell balling-up is cultivated with other cell direct contact type altogether
Loaded down with trivial details separating step after cultivation, and separate halfway interference；2, due to balling-up body constitution
Amount is relatively big, overcomes the factor that cannot get rid of gravity interference during Transwell co-cultures；3, suitable
Wide by property, it is not only suitable for tumor balling-up and cultivates and the co-culturing of attached cell, be also applied for tumor
Balling-up cultivation co-cultures with suspension cell；4, can be with low cost, simple with Reusability
Easily operation.This co-culture device is utilized to carry out tumor cell balling-up cultivation and cell-cell interaction
Effect observation, practicality simple to operate, effect are simple and clear.
Accompanying drawing explanation
In order to be illustrated more clearly that this utility model embodiment or technical scheme of the prior art,
The accompanying drawing used required in embodiment or description of the prior art will be briefly described below,
It should be evident that the accompanying drawing in describing below is only embodiments more of the present utility model, right
From the point of view of those of ordinary skill in the art, on the premise of not paying creative work, also may be used
To obtain other accompanying drawing according to these accompanying drawings.
Fig. 1 is overall structure schematic diagram of the present utility model；
Fig. 2 is the structural representation of single culture hole of the present utility model；
Fig. 3 is the assembling structural representation of single culture hole of the present utility model.
Detailed description of the invention
Below in conjunction with the accompanying drawing in this utility model embodiment, in this utility model embodiment
Technical scheme be clearly and completely described, it is clear that described embodiment be only this
The a part of embodiment of utility model rather than whole embodiments.Based in this utility model
Embodiment, those of ordinary skill in the art are obtained under not making creative work premise
Every other embodiment, any modification, equivalent substitution and improvement etc. made, should be included in
Within protection domain of the present utility model.
As shown in Figure 1, 2, 3, cell of the present utility model is cultivated and cell-cell interaction
Co-culture device, including culture plate 1, at least provided with a culture hole on described culture plate 1
2, culture hole 2 is divided into the detachable semi-transparent of several cell culture chambers 3,4 by least one
Film shaping piece 6 and be positioned at detachable semipermeable membrane partition 6 both sides detachable shaping piece 7a,
7b；It is arranged with in parallel vertically on the hole wall of described culture hole 2 and sets detachable semipermeable membrane for card
Shaping piece 6, draw-in groove 5a, 5b, 5c, 5a', 5b of detachable shaping piece 7a, 7b ', 5c ',
Described draw-in groove 5a, 5b, 5c, 5a', 5b ', the longitudinally opposed distribution of 5c '；Described draw-in groove 5a,
5b, 5c, 5a', 5b ', the height of 5c ' identical with the degree of depth of culture hole 2, described removable
From insertion draw-in groove 5c, 5c that semipermeable membrane shaping piece 6 is detachable ' in, detachable shaping piece 7a
In detachable insertion draw-in groove 5a, 5a', detachable insertion draw-in groove detachable for shaping piece 7b
5b, 5b ' in；Described culture hole 2 and detachable semipermeable membrane shaping piece 6, detachable semipermeable membrane
The junction of shaping piece 7a, 7b is provided with prevent liquid between cell culture chamber from flowing mutually close
Seal apparatus, detachable semipermeable membrane shaping piece 6 and draw-in groove 5c, 5c ' junction, detachable
Shaping piece 7a and the junction of draw-in groove 5a, 5a', detachable shaping piece 7b and draw-in groove 5b, 5b '
Junction be provided with the sealing device preventing liquid between cell culture chamber from flowing mutually.
Described detachable shaping piece 7a, 7b are symmetrically distributed in detachable semipermeable membrane shaping piece 6
Both sides, the most adjacent with detachable semipermeable membrane shaping piece 6, at a distance of width≤0.5mm；Described
Detachable semipermeable membrane shaping piece 6 is suitable to culture medium or that cell passes is semi-transparent for dismountable
Film shaping piece；Described detachable semipermeable membrane shaping piece 6 is polycarbonate membrane shaping piece；Described can
The pore size separating semipermeable membrane shaping piece 6 is 3-8 μm；Described detachable shaping piece 7a,
7b is the plastic sheet of dismountable acellular adhesion, and height is identical with the culture hole degree of depth.
Described culture plate 1 is 6 culture hole or 12 culture hole or the cell of 24 culture hole
Culture plate or Tissue Culture Dish, shown in Fig. 1 for 6 culture hole.
This cell cultivates the using method of the co-culture device with cell-cell interaction:
S1, co-culture device is carried out autoclaving and ultra-vioket radiation sterilizing；
S2, insert detachable semipermeable membrane shaping piece 6, detachable shaping piece 7a, 7b；
Under S3, sterile working, tumor cell suspension is added the cell cultivation being coated matrigel
Room 3, makes tumor cell be sunken to matrigel, stablizes grown cultures 3 in cell culture chamber 3
My god；
S4, the 4th day time, be coated another cell culture chamber 4 with poly-D-lysine, another kind of
Attached cell is planted in the cell culture chamber 4 being coated poly-D-lysine.37 DEG C of incubated overnight,
It is allowed to adherent growth in cell culture chamber 4；
S5, separate detachable shaping piece 7a, 7b, make tumor cell and attached cell co-culture；
S6, persistently co-culture, observe tumor cell balling-up situation, and the tumor cell of balling-up
From a cell culture chamber 3 to the migration situation of another cell culture chamber 4；The balling-up migrated
Tumor cell is attached on detachable semipermeable membrane shaping piece 6, it is simple to balling-up is observed in follow-up dyeing
The migration of tumor.
Cell of the present utility model is cultivated and the co-culture device of cell-cell interaction, and 1, knot
Structure is simple and practical, it is to avoid conventional tumor cell balling-up is cultivated with other cell direct contact type altogether
Loaded down with trivial details separating step after cultivation, and separate halfway interference；2, due to balling-up body constitution
Amount is relatively big, overcomes the factor that cannot get rid of gravity interference during Transwell co-cultures；3, suitable
Wide by property, it is not only suitable for tumor balling-up and cultivates and the co-culturing of attached cell, be also applied for tumor
Balling-up cultivation co-cultures with suspension cell；4, can be with low cost, simple with Reusability
Easily operation.This co-culture device is utilized to carry out tumor cell balling-up cultivation and cell-cell interaction
Effect observation, practicality simple to operate, effect are simple and clear.
Claims (9)
1. cell is cultivated and a co-culture device for cell-cell interaction, including culture plate,
It is characterized in that: cultivate at least provided with a culture hole, at least one general on described culture plate
Hole is divided into the detachable semipermeable membrane shaping piece of several cell culture chambers and is positioned at detachable
The detachable shaping piece of semipermeable membrane partition both sides.
Cell the most according to claim 1 cultivation co-cultures with cell-cell interaction
Device, it is characterised in that: it is arranged with in parallel vertically on the hole wall of described culture hole and sets for card
Detachable semipermeable membrane shaping piece, the draw-in groove of detachable shaping piece, the longitudinally opposed distribution of described draw-in groove；
The height of described draw-in groove is identical with the degree of depth of culture hole, described detachable semipermeable membrane shaping piece, can
Separate in the insertion draw-in groove that shaping piece is detachable.
Cell the most according to claim 2 cultivation co-cultures with cell-cell interaction
Device, it is characterised in that: described culture hole and detachable semipermeable membrane shaping piece, detachable semi-transparent
The junction of film shaping piece, draw-in groove and detachable semipermeable membrane shaping piece, the isolation of detachable semipermeable membrane
The junction of sheet is provided with the sealing device preventing liquid between cell culture chamber from flowing mutually.
Cell the most according to claim 1 cultivation co-cultures with cell-cell interaction
Device, it is characterised in that: described detachable shaping piece is symmetrically distributed in the isolation of detachable semipermeable membrane
The both sides of sheet, the most adjacent with detachable semipermeable membrane shaping piece, at a distance of width≤0.5mm.
Cell the most according to claim 1 cultivation co-cultures with cell-cell interaction
Device, it is characterised in that: described detachable semipermeable membrane shaping piece is that dismountable being suitable to cultivates Jie
The semipermeable membrane shaping piece that matter or cell pass.
Cell the most according to claim 5 cultivation co-cultures with cell-cell interaction
Device, it is characterised in that: described detachable semipermeable membrane shaping piece is polycarbonate membrane shaping piece.
Cell the most according to claim 6 cultivation co-cultures with cell-cell interaction
Device, it is characterised in that: the pore size of described detachable semipermeable membrane shaping piece is 3-8 μm.
Cell the most according to claim 1 cultivation co-cultures with cell-cell interaction
Device, it is characterised in that: described detachable shaping piece is moulding of dismountable acellular adhesion
Tablet, height is identical with the culture hole degree of depth.
Cell the most according to claim 1 cultivation co-cultures with cell-cell interaction
Device, it is characterised in that: described culture plate is 6 culture hole or 12 culture hole or 24 culture hole
Tissue Culture Plate or Tissue Culture Dish.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201620298442.3U CN205473829U (en) | 2016-04-11 | 2016-04-11 | Interact's culture apparatus between cell culture and cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201620298442.3U CN205473829U (en) | 2016-04-11 | 2016-04-11 | Interact's culture apparatus between cell culture and cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN205473829U true CN205473829U (en) | 2016-08-17 |
Family
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201620298442.3U Expired - Fee Related CN205473829U (en) | 2016-04-11 | 2016-04-11 | Interact's culture apparatus between cell culture and cell |
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| Country | Link |
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| CN (1) | CN205473829U (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105695328A (en) * | 2016-04-11 | 2016-06-22 | 张姝 | Co-culture device for cell culture and cell-cell interaction and using method of co-culture device |
| CN106754358A (en) * | 2016-12-23 | 2017-05-31 | 青岛农业大学 | A kind of co-culture of cells blake bottle of convenient transfer cell |
-
2016
- 2016-04-11 CN CN201620298442.3U patent/CN205473829U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105695328A (en) * | 2016-04-11 | 2016-06-22 | 张姝 | Co-culture device for cell culture and cell-cell interaction and using method of co-culture device |
| CN106754358A (en) * | 2016-12-23 | 2017-05-31 | 青岛农业大学 | A kind of co-culture of cells blake bottle of convenient transfer cell |
| CN106754358B (en) * | 2016-12-23 | 2023-06-30 | 青岛农业大学 | A kind of cell co-cultivation flask convenient for transferring cells |
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| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160817 Termination date: 20190411 |