CN109749979A - A kind of primary cell method for separating - Google Patents
A kind of primary cell method for separating Download PDFInfo
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- CN109749979A CN109749979A CN201910136039.9A CN201910136039A CN109749979A CN 109749979 A CN109749979 A CN 109749979A CN 201910136039 A CN201910136039 A CN 201910136039A CN 109749979 A CN109749979 A CN 109749979A
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Abstract
The invention belongs to cell sorting techniques fields, more particularly to a kind of primary cell method for separating, if this primary cell sorting method includes: to contain N kind cell in the primary cell chosen, it then needs to be correspondingly arranged on N-1 cell sorting devices, and above-mentioned cell sorting devices are controlled by controller, to be sorted to primary cell；This primary cell sorting method, sorts primary cell using cell sorting system, is able to achieve the sorting of primary cell.
Description
Technical field
The invention belongs to technical field of cell biology, and in particular to a kind of primary cell method for separating.
Background technique
When cultivating primary cell, because the extraction of primary cell is often mixed with other cells, one is caused to cultivation results
Fixed influence at this time must come out other cell sortings.
Therefore, a kind of primary cell method for separating is designed to be necessary.
Summary of the invention
The object of the present invention is to provide a kind of primary cell method for separating.
In order to solve the above-mentioned technical problems, the present invention provides a kind of primary cell method for separating, comprising:
If containing N kind cell in the primary cell chosen, need to be correspondingly arranged on N-1 cell sorting devices, and by controller
Above-mentioned cell sorting devices are controlled, to be sorted to primary cell.
Further, if N is 2 kinds of cells, two kinds of primary cells are sorted using a cell sorting devices；
The cell sorting devices include: culture dish, are provided with vibrator below the culture dish；Wherein
The vibrator and controller are electrically connected；
The side wall of the culture dish offers water outlet, and water outlet is provided with outlet valve；
The controller control vibrator vibration, so that non-attached cell floats in culture dish；
After the liquid level of culture solution is higher than water outlet in culture dish, the controller control outlet valve is opened, to release containing not
The culture solution of attached cell.
Further, the culture dish is additionally provided with water inlet, and the water inlet is provided with inlet valve；
After controller control inlet valve is opened, culture solution is supplemented into culture dish from water inlet；
Liquidometer is provided in the culture dish, the liquidometer is connected with controller；And
The controller by liquidometer control inlet valve and outlet valve aperture so that in culture dish the liquid level of culture solution with go out
The upper of the mouth of a river keeps approximate fair along height.
Further, if N is 3 kinds of cells or more, grade setting will be connected using multiple cell sorting devices, to a variety of primary
Cell is gradually sorted；
The cell sorting devices include: culture dish, are provided with vibrator below the culture dish；Wherein
The vibrator and controller are electrically connected；
The side wall of the culture dish offers water outlet, and water outlet is provided with outlet valve；
The controller control vibrator vibration, so that non-attached cell floats in culture dish；
After the liquid level of culture solution is higher than water outlet in culture dish, the controller control outlet valve is opened, to release containing not
The culture solution of attached cell is into the culture dish of next cell sorting devices.
Further, the culture dish is additionally provided with water inlet, and the water inlet is provided with inlet valve；
After controller control inlet valve is opened, culture solution is supplemented into culture dish from water inlet；
Liquidometer is provided in the culture dish, the liquidometer is connected with controller；And
The controller by liquidometer control inlet valve and outlet valve aperture so that in culture dish the liquid level of culture solution with go out
The upper of the mouth of a river keeps approximate fair along height.
Further, different according to the various cells adherent time, controller controls vibrator in corresponding cell sorting devices
Induction Peried；I.e.
Non- attached cell floats in this grade of culture dish after the corresponding cell sorting devices vibration of previous stage；
After the liquid level of culture solution in the culture dish of previous stage is higher than water outlet, controller control outlet valve is opened, with to
Next stage cell sorting devices are released the culture solution containing non-attached cell, and attached cell is retained；
It gradually sorts, so as to retain same type cell in the culture dish in corresponding cell sorting devices, realizes cell sorting.
The invention has the advantages that a kind of primary cell method for separating of the invention, by being set below culture dish
It is equipped with vibrator, the side wall of culture dish offers water outlet, water outlet is provided with outlet valve, controller controls vibrator vibration
It is dynamic, so that non-attached cell floats in culture dish；After the liquid level of culture solution is higher than water outlet in culture dish, controller is controlled out
Water valve is opened, with the culture solution containing non-attached cell of releasing.This primary cell sorting method is easy to operate, can sub-elect primary
Other cells in cell.
Detailed description of the invention
Present invention will be further explained below with reference to the attached drawings and examples.
Fig. 1 is the schematic diagram of cell sorting devices of the present invention；
Fig. 2 is the schematic diagram of company's stage arrangement of cell sorting devices of the present invention.
In figure:
Vibrator 1, culture dish 2, water inlet 201, water outlet 202, water pump 3.
Specific embodiment
In conjunction with the accompanying drawings, the present invention is further explained in detail.These attached drawings are simplified schematic diagram, only with
Illustration illustrates basic structure of the invention, therefore it only shows the composition relevant to the invention.
Embodiment 1
Fig. 1 is the schematic diagram of cell sorting devices of the present invention；
As shown in Figure 1, a kind of primary cell method for separating is present embodiments provided, using a cell sorting devices to two kinds of originals
It is sorted for cell, the cell sorting devices include: culture dish 2, and the lower section of the culture dish 2 is provided with vibrator 1；Its
Described in vibrator 1 and controller be electrically connected；The side wall of the culture dish 2 offers water outlet 202, and in water outlet
Outlet valve is provided at 202；The controller control vibrator 1 vibrates, so that non-attached cell floats in culture dish 2；It is training
The liquid level of culture solution in ware 2 is supported higher than after water outlet 202, the controller control outlet valve is opened, to release containing not adherent
The culture solution of cell.
In the present embodiment, the frequency of the initial time of vibration and duration and vibration can be according to the class of cell
Type is adjusted, such as starts to vibrate after cell starts culture 3 hours, and vibration duration can be at 1 minute, vibration frequency
It can be 30-60HZ range.
Further, the culture dish 2 is additionally provided with water inlet 201, is provided with inlet valve at the water inlet 201；Work as control
After device control inlet valve processed is opened, culture solution is supplemented into culture dish 2 from water inlet 201, to guarantee when unattended,
Culture solution in culture dish 2, which is able to maintain cell, to be continued to cultivate growth；Improve the automation degree of sorting of cell sorting system.
In the present embodiment, liquidometer is provided in the culture dish 2, the liquidometer is connected with controller；And it is described
Controller controls the aperture of inlet valve and outlet valve by liquidometer, so that the liquid level of culture solution and water outlet 202 in culture dish 2
It is upper keep approximate along height and maintain an equal level so that the non-attached cell to suspend as much as possible can flow out from water outlet 202, improve
The efficiency of separation.
The controller can have several I/O ports using the industrial control board with embeded processor, can be with vibration
Device is connected, and vibrator can be, but not limited to using vibrating motor；And this industrial control board also has display interface and USB interface, can
For connecting display and keyboard to carry out relevant parameter setting.
Such as: it can be mixed with a small amount of fibroblast in cultured myocardial, need to sub-elect fibroblast at this time,
And fibroblast it is adherent speed it is faster than cardiac muscle cell, after fibroblast is adherent, controller control vibrator vibration, with
2 cardiac myocyte of culture dish is set to float；After controller controls inlet valve opening, culture is supplemented into culture dish 2 from water inlet 201
Liquid；After the liquid level of culture solution in culture dish 2 is higher than water outlet 202, controller controls outlet valve and opens, to release containing cardiac muscle
The culture solution of cell, retains fibroblast, and the culture solution containing cardiac muscle cell that will release collects for using.
If containing N kind cell in the primary cell chosen, need to be correspondingly arranged on N-1 cell sorting devices.This implementation
In example, N is 2.Then controller control vibrator vibration, so that non-attached cell floats in culture dish 2；It is trained in culture dish 2
After the liquid level of nutrient solution is higher than water outlet 202, the controller control outlet valve is opened, with the culture containing non-attached cell of releasing
Liquid retains attached cell.
In the present embodiment, primary cell method for separating is only applicable to an adherent type primary cell either adherent type cell
With the sorting of a non-adherent type cell.
Embodiment 2
Fig. 2 is the schematic diagram of company's stage arrangement of cell sorting devices of the present invention.
As shown in Fig. 2, on the basis of embodiment 1, a kind of primary cell method for separating is present embodiments provided, if cell
Screening plant is two or more, then using even grade setting, the i.e. water outlet 202 of the cell sorting devices of previous stage and rear stage
The water inlet 201 of cell sorting devices is connected to, by the cell screening of the cell delivery of the non-attached cell of previous stage to rear stage
It is cultivated in device, realizes cell sorting.
Wherein, the water inlet 201 of the cell sorting devices of the water outlet 202 and rear stage of the cell sorting devices of previous stage
It is connected to by a water pump 3, so that the water outlet 202 of the low level of culture dish 2 will be not adherent in the cell sorting devices of previous stage
The high-order water inlet 201 of the cell smooth delivery of power of cell culture dish 2 into the cell sorting devices of rear stage.
In the present embodiment, N is 3 or more.Different according to the various cells adherent time, controller controls corresponding cell screening
The Induction Peried of vibrator 1 in device；Non- attached cell in this grade of culture dish 2 i.e. after the corresponding cell sorting devices vibration of previous stage
It floats；After the controller control inlet valve of the previous stage cell sorting devices is opened, mended from water inlet 201 into culture dish 2
Fill culture solution；After the liquid level of culture solution in the culture dish 2 of previous stage is higher than water outlet 202, the controller controls outlet valve
It opens, to release the culture solution containing non-attached cell to next stage cell sorting devices, retains attached cell；Gradually divide
Choosing realizes cell sorting so as to accordingly retain same type cell in the culture dish 2 in cell sorting devices.
In the present embodiment, various cells are different according to different tissue acquisition methods, and the adherent time of cell has any different, and receive
The type and concentration difference of digestive ferment (clostridiopetidase A and trypsase etc.) when collecting cell will also result in the adherent time and have any different,
Therefore the numerical value that the various cells adherent time in the present embodiment does not determine, as long as sorting different cells all in the present invention
Protection scope in.
In the present embodiment, primary cell method for separating is only applicable to adherent type primary cell.
In conclusion a kind of primary cell method for separating of the invention, by being provided with vibrator below culture dish,
The side wall of culture dish offers water outlet, water outlet is provided with outlet valve, controller controls vibrator vibration, so that culture
Non- attached cell floats in ware；After the liquid level of culture solution is higher than water outlet in culture dish, controller controls outlet valve and opens, with
It releases the culture solution containing non-attached cell, this primary cell sorting method is easy to operate, can sub-elect its in primary cell
His cell；This primary cell sorting method can be arranged N-1 cell sieve and choose to install according to N kind cell is contained in primary cell
It sets, various kinds of cell can be sorted, have the advantages that screening cell is practical.
Taking the above-mentioned ideal embodiment according to the present invention as inspiration, through the above description, relevant staff is complete
Various changes and amendments can be carried out without departing from the scope of the technological thought of the present invention' entirely.The technology of this invention
Property range is not limited to the contents of the specification, it is necessary to which the technical scope thereof is determined according to the scope of the claim.
Claims (6)
1. a kind of primary cell method for separating, comprising:
If containing N kind cell in the primary cell chosen, need to be correspondingly arranged on N-1 cell sorting devices, and by controller
Above-mentioned cell sorting devices are controlled, to be sorted to primary cell.
2. primary cell method for separating as described in claim 1, which is characterized in that
If N is 2 kinds of cells, two kinds of primary cells are sorted using a cell sorting devices；
The cell sorting devices include: culture dish, are provided with vibrator below the culture dish；Wherein
The vibrator and controller are electrically connected；
The side wall of the culture dish offers water outlet, and water outlet is provided with outlet valve；
The controller control vibrator vibration, so that non-attached cell floats in culture dish；
After the liquid level of culture solution is higher than water outlet in culture dish, the controller control outlet valve is opened, to release containing not
The culture solution of attached cell.
3. primary cell method for separating as claimed in claim 2, which is characterized in that
The culture dish is additionally provided with water inlet, and the water inlet is provided with inlet valve；
After controller control inlet valve is opened, culture solution is supplemented into culture dish from water inlet；
Liquidometer is provided in the culture dish, the liquidometer is connected with controller；And
The controller by liquidometer control inlet valve and outlet valve aperture so that in culture dish the liquid level of culture solution with go out
The upper of the mouth of a river keeps approximate fair along height.
4. primary cell method for separating as described in claim 1, which is characterized in that
If N is 3 kinds of cells or more, grade setting will be connected using multiple cell sorting devices, with to a variety of primary cells carry out by
Secondary sorting；
The cell sorting devices include: culture dish, are provided with vibrator below the culture dish；Wherein
The vibrator and controller are electrically connected；
The side wall of the culture dish offers water outlet, and water outlet is provided with outlet valve；
The controller control vibrator vibration, so that non-attached cell floats in culture dish；
After the liquid level of culture solution is higher than water outlet in culture dish, the controller control outlet valve is opened, to release containing not
The culture solution of attached cell is into the culture dish of next cell sorting devices.
5. primary cell method for separating as claimed in claim 4, which is characterized in that
The culture dish is additionally provided with water inlet, and the water inlet is provided with inlet valve；
After controller control inlet valve is opened, culture solution is supplemented into culture dish from water inlet；
Liquidometer is provided in the culture dish, the liquidometer is connected with controller；And
The controller by liquidometer control inlet valve and outlet valve aperture so that in culture dish the liquid level of culture solution with go out
The upper of the mouth of a river keeps approximate fair along height.
6. primary cell method for separating as claimed in claim 5, which is characterized in that
Different according to the various cells adherent time, controller controls the Induction Peried of vibrator in corresponding cell sorting devices；I.e.
Non- attached cell floats in this grade of culture dish after the corresponding cell sorting devices vibration of previous stage；
After the liquid level of culture solution in the culture dish of previous stage is higher than water outlet, controller control outlet valve is opened, with to
Next stage cell sorting devices are released the culture solution containing non-attached cell, and attached cell is retained；
It gradually sorts, so as to retain same type cell in the culture dish in corresponding cell sorting devices, realizes cell sorting.
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| CN201910136039.9A CN109749979B (en) | 2019-02-25 | 2019-02-25 | Primary cell sorting method |
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| CN201910136039.9A CN109749979B (en) | 2019-02-25 | 2019-02-25 | Primary cell sorting method |
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| CN109749979A true CN109749979A (en) | 2019-05-14 |
| CN109749979B CN109749979B (en) | 2020-11-27 |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE10223536A1 (en) * | 2002-05-27 | 2003-12-18 | J Hinrich Peters | Separation of non-adherent cells from adherent cells comprises rotating a culture vessel so that the non-adherent cells sediment away from the adherent cells under gravity |
| CN103484365A (en) * | 2013-08-30 | 2014-01-01 | 中国人民解放军南京军区福州总医院 | Device and method for obtaining stem cells from tissues |
| CN104694474A (en) * | 2015-03-31 | 2015-06-10 | 南京新诺丹生物技术有限公司 | Cell culture method |
| CN106047707A (en) * | 2016-08-03 | 2016-10-26 | 广州赛泊泰生物技术有限公司 | Wall adherence/suspension type cell culture unit as well as device, system and method thereof |
| CN108865654A (en) * | 2018-06-29 | 2018-11-23 | 苏州百源基因技术有限公司 | A kind of cell sorting device and method for separating |
-
2019
- 2019-02-25 CN CN201910136039.9A patent/CN109749979B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE10223536A1 (en) * | 2002-05-27 | 2003-12-18 | J Hinrich Peters | Separation of non-adherent cells from adherent cells comprises rotating a culture vessel so that the non-adherent cells sediment away from the adherent cells under gravity |
| CN103484365A (en) * | 2013-08-30 | 2014-01-01 | 中国人民解放军南京军区福州总医院 | Device and method for obtaining stem cells from tissues |
| CN104694474A (en) * | 2015-03-31 | 2015-06-10 | 南京新诺丹生物技术有限公司 | Cell culture method |
| CN106047707A (en) * | 2016-08-03 | 2016-10-26 | 广州赛泊泰生物技术有限公司 | Wall adherence/suspension type cell culture unit as well as device, system and method thereof |
| CN108865654A (en) * | 2018-06-29 | 2018-11-23 | 苏州百源基因技术有限公司 | A kind of cell sorting device and method for separating |
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