CN105603040A - Method used for observing mutual influences of PC12 cells at different physiological status - Google Patents
Method used for observing mutual influences of PC12 cells at different physiological status Download PDFInfo
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- CN105603040A CN105603040A CN201510762935.8A CN201510762935A CN105603040A CN 105603040 A CN105603040 A CN 105603040A CN 201510762935 A CN201510762935 A CN 201510762935A CN 105603040 A CN105603040 A CN 105603040A
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Abstract
The invention discloses a method used for observing mutual influences of PC12 cells at different physiological status. The method comprises following steps: a PC12 cell strain steadily transfected by GFP is constructed; co-culture systems of infected PC12 cells and non-infected PC12 cells are constructed; different dyes are added, and ROS level, apoptosis level, and mitochondrial membrane potential of the co-culture systems are detected. Direct contact mixing co-culturing is adopted; after co-culturing of lead acetate infected PC12 cells and non-infected PC12 cells, the different dyes are added, the base color of infected PC12 cells is fluorescent green, and fluorescence overlaying with other colors can be realized, so that the infected PC12 cells can be distinguished, it can be observed whether the normal physiological states of surrounding non-infected PC12 cells are influenced by infected PC12 cells, and the different physiological states of PC12 cells under a same culturing environment are distinguished effectively.
Description
Technical field
The invention belongs to cell biology field, more specifically, the present invention relates to a kind of different physiology shapes of observingInteractional method between state PC12 cell.
Background technology
The microenvironment of Growth of Cells be keep that cell is normally bred, differentiation, metabolism and functional activity importantPart. Cell microenvironment is to external world very responsive, in same co-culture system, between cell or cell andDirect contact between extracellular matrix can affect the growth conditions of cell. In recent decades, at radiation biologicalField is found, in the time that a few cell is subject to radiation exposure, is not subject to the peripheral cell of ray direct irradiationAlso can show some damage effects.
Large quantity research has proved really to have damage transmission effects in vivo with in experiment in vitro. For example, slowProperty granulocytic leukemia infant spleen in the time accepting radiation exposure treatment, there is damage in breastbone marrow cellWound; Selective oxidation damage can be transmitted by GJIC between endothelial cell. Someone proposes with the tumour base of committing suisideBecause for the treatment of tumour, when after Partial tumors cell transfecting suicide gene, start after transfectional cell suicide at medicine,Transfectional cell and around non-transfected cells are all killed. In recent years, there is human hair current situation portion oxidative damage to lead toCross Intercellular Bridge connecting communication (GJIC) transmits between epithelial cell. The above exists one jointlyPhenomenon, be exactly that the target cell damage that causes of some adverse factor is not only confined to target cell itself, also canTo cause that surrounding normal cell produces damage. The cell mass that is exposed to adverse factor there will be obvious biological effectShould, this effect exposed cell group and around not exposed cell group all can occur, this phenomenon people are referred to asBystander effect (bystandereffect, BE).
Bystander effect refers to what normal cell occurred under the target cell induction that is directly subject to adverse factor infringementSuch as reactions such as cell death, gene mutation, chromosome instability are fixed, through bringing out, generating effect is being just for theseNormal cell becomes bystander cell line. Due to the existence of bystander effect, the damage effect of target cell is prolongedContinue and amplify, thereby damage is obviously increased the weight of. People are reminded in these researchs: body to the reaction of damage not onlyBe only single independent cell or tissue, but there is group. Discovery for bystander effect is existing several10 years, but up to now, the precise mechanism of bystander effect is not yet illustrated. More and more research shows, sideGJIC (the gap that onlooker's effect may mediate with inserted by connexin (connexin, Cx)-junctionalintercellularcommunication, GJIC) there is a substantial connection.
In external interior research, generally adopt the contamination of cultured cell entirety at present, adopt cBX inhibition gap to connectConnect, the in the situation that of damaging cells, detect the apoptosis situation of inducing cell, seek the letter in damage processNumber pass through mechanism, but this kind of method fails effectively distinguish contamination and do not contaminate cell. Somebody is shone by radiationPenetrate the part cell in culture dish, observe the not apoptosis situation of illuminated cell, but this research method is only limitIn radioactive exposure, fail to be applied to the impact of the chemical substance being dissolved in nutrient solution on cell. How effectively districtDividing contamination in co-cultured cell is the technical barrier of being badly in need of solution with the cell of not contaminating.
Summary of the invention
In view of this,, in order to overcome above-mentioned problems of the prior art, the invention provides a kind of observationInteractional method between different physiological status PC12 cells, it is thin that the method can be distinguished different physiological statusBorn of the same parents under same culture environment, contamination with do not contaminate the physiological status of PC12 cell.
In order to realize foregoing invention object, the present invention has taked following technical scheme:
Interactional method between the different physiological status PC12 cells of a kind of observation, comprises the following steps:
(1), build GFP stable transfection PC12 cell line
PC12 cell in exponential phase is discarded to original fluid, appropriate with adding after PBS washingTrypsin-EDTA, is seeded to six orifice plates after digestion, after cell is completely adherent, transduces, and before transduction,Every porocyte changes fresh HD+10%FBS culture medium or 1640+20%FBS culture medium into, and every strain chooses onePorocyte adds the Lenti-eGFP/Neo of 30 μ L, fully shakes up in rear incubator and cultivates transduction after 7 hours, inhalesRemove the culture medium that contains Lenti-eGFP/Neo, change into fresh HD+10%FBS culture medium or1640+20%FBS culture medium, then put into incubator and cultivate transduction 48 hours, build and obtain GFP-PC12;
(2), co-culture system is set up
Two co-culture systems are set: the one, in GFP-PC12, add virulence factor cultivate after, by PC12 withThe GFP-PC12 processing through MPb mixes altogether and cultivates by equal proportion; The 2nd, by PC12 and GFP-PC12 by geometric ratioExample is mixed altogether and is cultivated, and two cultivating systems are all cultivated after 20 minutes altogether in 37 DEG C, rinses cell 2～3 times with PBS;
(3), the ROS level of co-culture system, level of apoptosis, mitochondrial transmembrane potentials detect
In two co-culture systems, add respectively Hoechst33342 dyestuff to carry out core and redye, laser co-focusing dividesAnalyse ROS level in cell;
In two co-culture systems, add TMRM to carry out mitochondrial transmembrane potentials detection respectively;
In two co-culture systems, add respectively AnnexinV-PE or 7-AAD to carry out Level of Apoptosis inspectionSurvey.
In some embodiment, in step (1), after described PC12 cell dissociation, press 1 × 10 therein5cells/9.6cm2Density be seeded on six orifice plates.
In some embodiment, described in step (1), the condition of culture of incubator is therein: 37 DEG C, 5%CO2、95% relative humidity.
In some embodiment, in step (2), cultivating the culture medium using is altogether HD+10%FBS culture medium thereinOr 1640+20%FBS culture medium.
In some embodiment, the ROS level in step (3) in cell is to use with 350nm excitation wave thereinLong, the fluorescence microscope of the optical filter of 460nm emission wavelength is analyzed.
In some embodiment, when in step (3), AnnexinV-PE carries out Level of Apoptosis detection, flow thereinThe excitation wavelength Ex=488nm of formula cell instrument, emission wavelength Em=578nm, uses FL2 passage to detect.
In some embodiment, when in step (3), 7-AAD carries out Level of Apoptosis detection, streaming is thin thereinThe excitation wavelength Ex=546nm of born of the same parents' instrument, emission wavelength Em=647nm, uses FL3 passage to detect.
In some embodiment, the concentration of Hoechst33342 dyestuff described in step (3) is 50 μ M therein.
Compared with prior art, the present invention has following beneficial effect:
Between the different physiological status PC12 cells of observation of the present invention, interactional method is contaminated lead acetateCell directly contacts and mixes after cultivation altogether with the cell of not contaminating, and adds different dyestuffs, due to contamination cell basedEnd color is green fluorescence, can superpose with different colours fluorescence, so just can distinguish contamination cell, therebyCan observe contamination cell whether the periphery normal cell normal physiological state of not contaminating has been caused to impact, effectivelyDistinguish the different physiological status of the cell under same culture environment, before having solved, can not effectively distinguish same trainingSupport the problem of the different contamination states of allogenic cell under environment.
Detailed description of the invention
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with concrete enforcementExample, is further elaborated to the present invention. Only should be appreciated that specific embodiment described hereinIn order to explain the present invention, be not intended to limit the present invention.
Raw material in following examples if no special instructions, all derives from commercially available.
Interactional method between 1 one kinds of different physiological status PC12 cells of observation of embodiment
Comprise following concrete steps:
(1) PC12 cell recovery
From liquid nitrogen container, take out PC12 cell cryopreservation tube, drop into rapidly in 37 DEG C of water-baths, and shake makes it as early as possibleMelt, after cryopreservation tube alcohol disinfecting, with bus suction pipe sucking-off cell suspending liquid, inject centrifuge tube and add 10 timesThe DMEM nutrient solution that contains 5% hyclone and 5% horse serum, mix after centrifugal (1000rpm, 5min),Remove supernatant, be diluted to 1 with nutrient solution (the DMEM nutrient solution that contains 5% hyclone and 5% horse serum)×104The cell suspension of individual/ml density, is inoculated in 25cm2In blake bottle, every bottle of 10ml, is placed in 37 DEG C, contains5％CO2Incubator in cultivate.
(2) PC12 cell is cultivated
PC12 cell is cultivated through recovery, after cell monolayer covers with, abandons culture medium, and every bottle adds 1ml0.25% pancreasEnzyme-EDTA digests, and adds containing the complete culture solution of 10% hyclone and stop disappearing after part cell detachmentChange, cell is blown and beaten gently and is dispersed to after single cell suspension, move in 10ml centrifuge tube, centrifugal after mixing(1000rpm, 5min), abandons supernatant, resuspended containing the complete culture solution of 10% serum with 1mL, average markBecome three parts, be inoculated in three 25cm2In blake bottle, be placed in incubator and cultivate by above-mentioned steps () method,Every 2-3 days goes down to posterity once. Test cell used all in exponential phase.
(3) stable cell strain builds
1, before PC12 cell transfecting, prepare
(1) by HD+10%FBS culture medium, 1640+20%FBS culture medium, 1 × PBS and Trypsin-EDTAPre-balance is to arriving room temperature;
(2) by aseptic the cell conditioned medium collection censorship of going down to posterity after cultivating, after aseptic testing result is determined, logicalCross the cultivation of going down to posterity (use HD+10%FBS culture medium or 1640+20%FBS culture medium) and carry out cell stateAdjust, cell sucks culture medium after covering with in blake bottle, with 1 × PBS washed cell 2-3 time, to removeResidual serum;
(3) suck PBS, add appropriate Trypsin-EDTA, rotation, makes Trypsin-EDTA even gentlyCovering culture vessel surface, digests and within 1-2 minute, makes cell take off wall, and under microscope, visible cell gap increases, thinIt is round that born of the same parents become, and claps the wall of culture vessel with have gentle hands, add immediately 2mL HD+10%FBS culture medium or1640+20%FBS culture medium stops digestion;
(4) use suction pipe imbitition, blow and beat gently culture vessel surface, 3-5 time repeatedly, make cell thoroughly de-From bottle ware diapire, cell is moved in centrifuge tube, then add 1 × PBS to wash 1-2 time in blake bottle, and will washLiquid is transferred in centrifuge tube in the lump;
(5) centrifugal 4 minutes of 244g, sucks supernatant;
(6) add the HD+10%FBS culture medium of 5mL or 1640+20%FBS culture medium re-suspended cell to precipitate,Blow and beat gently the cell suspension of collection with suction pipe, sampling counting;
(7), with reference to count results, get 1 × 105cells/9.6cm2Density be seeded to six orifice plates, respectively inoculate 2Individual hole.
2, GFP transduction PC12 cell
(1) after cell is completely adherent, can transduce; Before transduction, it is new that every porocyte changes 1.5mL intoFresh HD+10%FBS culture medium or 1640+20%FBS culture medium, every strain is chosen a porocyte and is added 30The Lenti-eGFP/Neo of μ L, puts into 37 DEG C, 5%CO after fully shaking up2, 95% relative humidity incubatorMiddle cultivation;
(2) transduction, after 7 hours, sucks the complete medium that contains Lenti-eGFP/Neo, and washes two with PBSInferior; Change fresh HD+10%FBS culture medium or 1640+20%FBS culture medium into, put into 37 DEG C, 5%CO2、In the incubator of 95% relative humidity, cultivate;
(3) transduction is after 48 hours, in fluorescence microscopy Microscopic observation transduction effect;
(4) after cell covers with, by the PC12/GFP-Neo amplification of going down to posterity. Transduce and after 48 hours, observe GFP and turnLead the transduction rate of PC12 up to 90%, and fluorescence is stronger.
3, PC12/GFP-Neo cell cryopreservation
In the time that cell grows to 80% degrees of fusion, by sampling counting after cell dissociation, according to count results, by 1 ×106Cells/vial carries out frozen.
(4) the ROS level of co-culture system, level of apoptosis, mitochondrial transmembrane potentials detect
Two co-culture systems are set: the one, in GFP-PC12, add MPb or ROS scavenger or gap to connectConnect after the virulence factor cultivations such as blocking agent, PC12 is mixed altogether by equal proportion with the GFP-PC12 processing through MPbCultivate; The 2nd, PC12 is mixed altogether and cultivated by equal proportion with GFP-PC12, two cultivating systems are all in 37 DEG C altogetherCultivate after 20 minutes, rinse cell 2～3 times with PBS;
Co-culture system method for building up step is:
(1), PC-12GFP (+) cell is with approximately 1 × 104Individual/mL density is inoculated in 24 orifice plates, every hole inoculation 500μl，5％CO2, 37 DEG C of overnight incubation, treat cell attachment;
(2), next day, the culture medium of PC-12GFP (+) cell is replaced by various experiment culture mediums (containing vinegarLead plumbate/ROS scavenger/gap connects blocking agent, cultivates 24h; PC-12GFP (-) is with approximately 0.5 × 104Individual/mLDensity is inoculated in 24 orifice plates, and 500 μ l, 5%CO are inoculated in every hole2, 37 DEG C of overnight incubation, treat cell attachment;
(3), by after PC-12GFP after treatment (+) trypsinization, be diluted to 1000-10000/mL, connectPlant to PC-12GFP (-), cultivate altogether 24-48h;
(4), absorb culture medium, after PBS washed cell 1-2 time, add 250 μ l cell culture mediums (freshHD+10%FBS culture medium or 1640+20%FBS culture medium);
Two co-culture systems are detected as follows:
1, ROS level in laser co-focusing analysis of cells
(1), inoculating cell: Trypsin-EDTA digestion exponential phase cell, be inoculated into 1% poly badOn the coated sterility cover slide of propylhomoserin, cell is put into incubator and leave standstill and cultivate, regularly change liquid;
(2), core redyes: drip Hoechest33342 solution carry out core redye after with 350nm excitation wavelength,The Confocal fluorescence microscopy Microscopic observation of the optical filter of 460nm emission wavelength is also taken a picture.
Hoechst33342 is blue-fluorescence, because color at the bottom of contamination cell based is green fluorescence, glimmering with bluenessOptical superposition, can distinguish contamination cell.
2, mitochondrial membrane potential detects (TMRM)
(1), add TMRM working solution, fully mix 5%CO2, cultivate 30-60min for 37 DEG C;
(2), 37 DEG C hatch after end, absorb supernatant, with PBS washed cell 2 times;
(3), add after the connection buffer solution after 400 μ l dilutions the inspection of cells were tested by flow cytometry mitochondrial membrane potentialSurvey.
TMRM is fluorescent orange, because color at the bottom of contamination cell based is green fluorescence, and with fluorescent orange stack,Can distinguish contamination cell.
3, flow cytometry analysis Apoptosis
(1), suspension cell centrifugal (the centrifugal 5min of 2000rpm) is collected; The pancreas that does not contain EDTA for attached cellEnzymic digestion collection (note: the trypsinization time is difficult for long, otherwise easily cause false positive);
(2), collect 1～5 × 10 with PBS washed cell secondary (the centrifugal 5min of 2000rpm)5Cell;
(3), add the BindingBuffer suspension cell of 500 μ L;
(4), add 1 μ LAnnexinV-PE to mix; Room temperature, lucifuge, reaction 5～15min;
(5), add 5 μ L7-AAD dye liquors, mix; Room temperature, lucifuge, reaction 5～15min;
(6), in 1hour, carry out observation and the detection of fluorescence microscope or flow cytometer.
Detect apoptosis rate, excitation wavelength Ex=488nm with flow cytometer; Emission wavelength Em=578nm,The fluorescent red-orange suggestion of AnnexinV-PE is used FL2 passage to detect; Excitation wavelength Ex=546nm; Transmitted waveLong Em=647nm, the suggestion of 7-AAD red fluorescence is used FL3 passage to detect. Fluorescence Compensation Regulation: use notThrough the normal cell of apoptosis induction processing, carry out in contrast fluorescence Compensation Regulation and remove spectra overlapping and settingThe position of cross door.
AnnexinV-PE is fluorescent red-orange, and the 7-AAD fluorescence that takes on a red color, because color at the bottom of contamination cell based isGreen fluorescence, with blue-fluorescence stack, can distinguish contamination cell.
The above embodiment has only expressed embodiments of the present invention, and it describes comparatively concrete and detailed, butCan not therefore be interpreted as the restriction to the scope of the claims of the present invention. It should be pointed out that for this areaThose of ordinary skill, without departing from the inventive concept of the premise, can also make some distortion and changeEnter, these all belong to protection scope of the present invention. Therefore, the protection domain of patent of the present invention should be with appended powerProfit requires to be as the criterion.
Claims (8)
1. observe an interactional method between different physiological status PC12 cells, it is characterized in that, bagDraw together following steps:
(1), build GFP stable transfection PC12 cell line
PC12 cell in exponential phase is discarded to original fluid, appropriate with adding after PBS washingTrypsin-EDTA, is seeded to six orifice plates after digestion, after cell is completely adherent, transduces, and before transduction,Every porocyte changes fresh HD+10%FBS culture medium or 1640+20%FBS culture medium into, and every strain chooses onePorocyte adds the Lenti-eGFP/Neo of 30 μ L, fully shakes up in rear incubator and cultivates transduction after 7 hours, inhalesRemove the culture medium that contains Lenti-eGFP/Neo, change into fresh HD+10%FBS culture medium or1640+20%FBS culture medium, then put into incubator and cultivate transduction 48 hours, build and obtain GFP-PC12;
(2), co-culture system is set up
Two co-culture systems are set: the one, in GFP-PC12, add virulence factor cultivate after, by PC12 withThe GFP-PC12 processing through MPb mixes altogether and cultivates by equal proportion; The 2nd, by PC12 and GFP-PC12 by geometric ratioExample is mixed altogether and is cultivated, and two cultivating systems are all cultivated after 20 minutes altogether in 37 DEG C, rinses cell 2～3 times with PBS;
(3), detect ROS level, level of apoptosis, the mitochondrial transmembrane potentials of co-culture system
In two co-culture systems, add respectively Hoechst33342 dyestuff to carry out core and redye, laser co-focusing dividesAnalyse ROS level in cell;
In two co-culture systems, add TMRM to carry out mitochondrial transmembrane potentials detection respectively;
In two co-culture systems, add respectively AnnexinV-PE or 7-AAD to carry out Level of Apoptosis inspectionSurvey.
2. interactional method between the different physiological status PC12 cells of observation according to claim 1,It is characterized in that, in step (1), after described PC12 cell dissociation, press 1 × 105cells/9.6cm2Density inoculationTo six orifice plates.
3. interactional method between the different physiological status PC12 cells of observation according to claim 1,It is characterized in that, in step (2), cultivate altogether the culture medium that uses for HD+10%FBS culture medium or1640+20%FBS culture medium.
4. interactional method between the different physiological status PC12 cells of observation according to claim 1,It is characterized in that, the ROS level in step (3) in cell is to use with 350nm excitation wavelength, and 460nm sends outThe fluorescence microscope of the optical filter of ejected wave length is analyzed.
5. interactional method between the different physiological status PC12 cells of observation according to claim 1,It is characterized in that, AnnexinV-PE carries out Level of Apoptosis while detecting in step (3), the swashing of flow cytometerSend out wavelength Ex=488nm, emission wavelength Em=578nm, uses FL2 passage to detect.
6. interactional method between the different physiological status PC12 cells of observation according to claim 1,It is characterized in that, when in step (3), 7-AAD carries out Level of Apoptosis detection, the excitation wave of flow cytometerLong Ex=546nm, emission wavelength Em=647nm, uses FL3 passage to detect.
7. interactional method between the different physiological status PC12 cells of observation according to claim 1,It is characterized in that, the concentration of Hoechst33342 dyestuff described in step (3) is 50 μ M.
8. interactional method between the different physiological status PC12 cells of observation according to claim 1,It is characterized in that, described in step (1), the condition of culture of incubator is: 37 DEG C, 5%CO2, 95% relatively wetDegree.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101848995A (en) * | 2007-06-25 | 2010-09-29 | 南达科他州立大学 | Recombinant north american type 1 porcine reproductive and respiratory syndrome virus and methods of use |
| CN103074300A (en) * | 2012-11-30 | 2013-05-01 | 甘肃中医学院 | Co-culture system establishment of mesenchymal stem cells and tumor cells as well as mesenchymal stem cells heredity stability change characteristic in tumor microenvironment |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101848995A (en) * | 2007-06-25 | 2010-09-29 | 南达科他州立大学 | Recombinant north american type 1 porcine reproductive and respiratory syndrome virus and methods of use |
| CN103074300A (en) * | 2012-11-30 | 2013-05-01 | 甘肃中医学院 | Co-culture system establishment of mesenchymal stem cells and tumor cells as well as mesenchymal stem cells heredity stability change characteristic in tumor microenvironment |
Non-Patent Citations (1)
| Title |
|---|
| 郭庶: "铅诱导PC12细胞旁观者效应及细胞间隙连接通讯（GJIC）作用研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
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Inventor after: Guo Shu Inventor after: Zhou Jin Inventor after: Yu Yunjiang Inventor after: Sun Jiaren Inventor after: Xiang Mingdeng Inventor after: Li Liangzhong Inventor after: Zhang Zongyao Inventor after: Tan Haiping Inventor after: Yang Jian Inventor before: Guo Shu |
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Effective date of registration: 20171019 Address after: 510535 No. 18, Rui Lu, Luogang District, Guangdong, Guangzhou Applicant after: South China Institute of Environmental Sciences. MEP Applicant after: GUANGZHOU WOMEN AND CHILDREN'S MEDICAL CENTER Address before: 510535 No. 18, Rui Lu, Luogang District, Guangdong, Guangzhou Applicant before: South China Institute of Environmental Sciences. MEP |
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Application publication date: 20160525 |