CN105950559A - Method for neuroblastoma differentiation on chiral gold nanoparticle film - Google Patents
Method for neuroblastoma differentiation on chiral gold nanoparticle film Download PDFInfo
- Publication number
- CN105950559A CN105950559A CN201610592436.3A CN201610592436A CN105950559A CN 105950559 A CN105950559 A CN 105950559A CN 201610592436 A CN201610592436 A CN 201610592436A CN 105950559 A CN105950559 A CN 105950559A
- Authority
- CN
- China
- Prior art keywords
- handed
- golden nanometer
- cell
- nanometer particle
- thin film
- Prior art date
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0693—Tumour cells; Cancer cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/05—Inorganic components
- C12N2500/10—Metals; Metal chelators
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/06—Anti-neoplasic drugs, anti-retroviral drugs, e.g. azacytidine, cyclophosphamide
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Oncology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Medicinal Preparation (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a method for neuroblastoma differentiation on a chiral gold nanoparticle film, and belongs to the technical field of biological science. The method includes the following steps of synthesizing gold nanoparticles with the particle size of 20 nm, preparing the single-layer gold nanoparticle film and culturing and inducing nerve cells in a differentiation mode. A chiral culture platform for cell selective differentiation is provided, and a new path is provided for treatment on related diseases caused by nervous damage.
Description
Technical field
The present invention relates to a kind of method that neuroblastoma is broken up on chiral gold nanoparticulate thin films, belong to bioscience technical field.
Background technology
Over nearly 20 years, the research of the molecular mechanism of neuroblastoma morbidity obtains remarkable progress, and Therapeutic Method also has many improvement, but clinical efficacy there is no much raisings.But, this peculiar phenomenon of the retrogression of nature of neuroblastoma but attracts people and pays close attention to.Neuroblastoma is considered as the tumor most being hopeful to disclose cancer eliminating natural process, contributes to finally taking measures to suppress its pernicious process.
Neuroblastoma can from row degradation, minority express the case of N-myc gene few or without systematic treating in the case of also can completely disappear or break up.Therefore people imagine the differentiation of inducing neural blastoma, and reversing its pernicious process may be than direct killing tumor cell more in accordance with nature.In vitro study finds, the material of energy inducing neural blastoma differentiation has retinoic acid, gamma interferon, dibutyl cyclic adenosine monophosphate, sodium phenylacetate etc., and wherein most study is retinoic acid.Retinoic acid has a high potential in terms of prevention and treatment cancer, and well-tolerated in Clinical practice, even if single cases is the most reactionless to radiotherapy, chemotherapy, with can long-term remission after retinoic acid treatment.
The research of induction differentiation based on neuroblastoma, people have done a lot of outstanding work.But, still there is the biggest exploration space at chirality interface on the differentiation impact of neuroblastoma.The present invention utilizes single layer of gold nanoparticulate thin films to build left-right rotary chirality interface, discloses chirality interface important function played in the differentiation of inducing neural blastoma.
Summary of the invention
It is an object of the invention to provide a kind of method that neuroblastoma is broken up on chiral gold nanoparticulate thin films.
Technical scheme, a kind of method that neuroblastoma is broken up on chiral gold nanoparticulate thin films, step is:
(1) neuroblastoma cultivation on chiral gold nanoparticulate thin films: left-handed or right-handed chirality golden nanometer particle thin film is placed in the cell culture plate well in 6 holes；With the neuroblastoma cell NG108-15 5min in 5mL trypsinization 25mL Tissue Culture Flask, after pancreatin is outwelled, by 1640 culture medium containing 20% hyclone, cell is blown afloat；Add containing 2 × 10 in being placed with the cell culture plate well of golden nanometer particle thin film of left-handed or right-handed chirality4The culture medium of individual cell/mL, is placed in 37 DEG C of CO2 gas incubator cultivation；Observation of cell form and density after 24h, on the golden nanometer particle thin film of left-handed chirality, cell is strip extended position, on the golden nanometer particle thin film of right-handed chirality, cell is rounded, and on the golden nanometer particle thin film of left-handed chirality, cell density is more than cell density on the golden nanometer particle thin film of right-handed chirality；
(2) neuroblastoma differentiation on chiral gold nanoparticulate thin films: after cell cultivates 24h on the left-handed and golden nanometer particle thin film of right-handed chirality respectively, the culture medium more renewed, add retinoic acid extremely final concentration of 1 μM, after continuing to cultivate 2 days, observation of cell form；On the left-handed and golden nanometer particle thin film of right-handed chirality, cell all grows synapse；Wherein, on the golden nanometer particle thin film of left-handed chirality, cell is slender type, and a two ends grows synapse, and on the golden nanometer particle thin film of right-handed chirality, surrounding all has synapse to grow.
The preparation process of described chiral monolayers golden nanometer particle thin film is as follows:
(1) golden nanometer particle of 20nm particle diameter synthesizes and L/D type cysteine modified:
The synthesis of a, 3-4nm gold nano seed: use sodium borohydride reduction synthesis；Take 120mL ultra-pure water, be sequentially added into 147 μ L mass concentration 1% trisodium citrates and 0.5mL 10mM gold chloride, after being sufficiently mixed, add the sodium borohydride of the 0.1M that 0.6mL newly prepares when being stirred vigorously, i.e. obtain 3-4nm gold nano seed solution；
The golden nanometer particle synthesis of b, 20nm particle diameter: use seed mediated growth method synthesis；5mL 10mM gold chloride is added in 195mL ultra-pure water, it is heated to boiling, it is simultaneously introduced 1.5mL gold nano seed solution and the trisodium citrate of 800 μ L 1% that step a prepares, continues to boil to color constant, i.e. obtain the solution of gold nanoparticles of 20nm particle diameter；
Synthetic solution of gold nanoparticles 9000rpm is centrifuged 10min, and 25 DEG C of concentrations are resuspended in the ultra-pure water of original solution 1/10 volume, regulate pH to 10 with sodium hydroxide, stand-by；Taking two parts of solution, be separately added into L-type cysteine and D type cysteine extremely final concentration of 100 μ g/mL, be stirred overnight, 9000rpm is centrifuged, and removes cysteine unnecessary in solution, resuspended, obtains the left-handed and solution of gold nanoparticles of right-handed chirality；
(2) preparation of golden nanometer particle thin film and transfer: the solution of gold nanoparticles of the left-handed or right-handed chirality that step (1) gained has been modified L/D type cysteine is poured in small beaker, adds 5mL normal hexane, is then slowly added dropwise dehydrated alcohol；During dropping, gold film can gradually on interface, be transferred on polydimethylsiloxane film by emersion by gold film, obtains having the left-handed or golden nanometer particle thin film of right-handed chirality, dries stand-by.
Beneficial effects of the present invention: the present invention utilizes single layer of gold nanoparticulate thin films to build left-right rotary chirality interface, discloses chirality interface important function played in the differentiation of inducing neural blastoma.
Accompanying drawing explanation
Fig. 1 is to transfer to the golden nanometer particle thin film on polydimethylsiloxane film in the present invention.
Fig. 2-a is the UV-vis absorption spectrum of left-handed golden nanometer particle thin film.
Fig. 2-b is the UV-vis absorption spectrum of dextrorotation golden nanometer particle thin film.
Fig. 3 is the circular dichroism collection of illustrative plates of chiral gold nanoparticulate thin films in the present invention.
Fig. 4 is the microphotograph after NG108-15 cell cultivates 24h on gold film.
Fig. 5 is to add Induced by Retinoic Acid laser co-focusing photo of noble cells on chirality golden nanometer particle thin film after 2 days.
Detailed description of the invention
Embodiment 1
1, the preparation of chiral monolayers golden nanometer particle thin film
The golden nanometer particle synthesis of (1) 20 nm particle diameter and L/D type cysteine modified；The golden nanometer particle of 20 nm particle diameters uses seed mediated growth method synthesis.
The synthesis of a, 3-4nm gold nano seed: use sodium borohydride reduction synthesis；Take 120mL ultra-pure water, be sequentially added into 147 μ L mass concentration 1% trisodium citrates and 0.5mL 10mM gold chloride, after being sufficiently mixed, add the sodium borohydride of the 0.1M that 0.6mL newly prepares when being stirred vigorously, i.e. obtain 3-4nm gold nano seed solution；
The golden nanometer particle synthesis of b, 20nm particle diameter: use seed mediated growth method synthesis；5mL 10mM gold chloride is added in 195mL ultra-pure water, it is heated to boiling, it is simultaneously introduced 1.5mL gold nano seed solution and the trisodium citrate of 800 μ L 1% that step a prepares, continues to boil to color constant, i.e. obtain the solution of gold nanoparticles of 20nm particle diameter；
Synthetic solution of gold nanoparticles 9000rpm is centrifuged 10min, and 25 DEG C of concentrations are resuspended in the ultra-pure water of original solution 1/10 volume, regulate pH to 10 with sodium hydroxide, stand-by；Taking two parts of solution, be separately added into L-type cysteine and D type cysteine extremely final concentration of 100 μ g/mL, be stirred overnight, 9000rpm is centrifuged, and removes cysteine unnecessary in solution, resuspended, obtains the left-handed and solution of gold nanoparticles of right-handed chirality；
(2) preparation of golden nanometer particle thin film and transfer: pour in small beaker by the solution of gold nanoparticles having modified L/D type cysteine, adds 5mL normal hexane, is then slowly added dropwise dehydrated alcohol.During dropping, gold film can gradually emersion on interface.Gold film can be transferred on PDMS film, and the golden nanometer particle thin film obtaining having left-handed (L-film) or dextrorotation (D-film) chirality dries stand-by.
2, neuroblastoma cultivation on chiral gold nanoparticulate thin films: left-handed or right-handed chirality golden nanometer particle thin film is placed in the cell culture plate well in 6 holes；With the neuroblastoma cell NG108-15 5min in 5mL trypsinization 25mL Tissue Culture Flask, after pancreatin is outwelled, by 1640 culture medium containing 20% hyclone, cell is blown afloat；Add containing 2 × 10 in being placed with the cell culture plate well of golden nanometer particle thin film of left-handed or right-handed chirality4The culture medium of individual cell/mL, is placed in 37 DEG C of CO2 gas incubator cultivation；Observation of cell form and density after 24h, on the golden nanometer particle thin film of left-handed chirality, cell is strip extended position, on the golden nanometer particle thin film of right-handed chirality, cell is rounded, and on the golden nanometer particle thin film of left-handed chirality, cell density is more than cell density on the golden nanometer particle thin film of right-handed chirality；
3, neuroblastoma differentiation on chiral gold nanoparticulate thin films: after cell cultivates 24h on the left-handed and golden nanometer particle thin film of right-handed chirality respectively, the culture medium more renewed, add retinoic acid extremely final concentration of 1 μM, after continuing to cultivate 2 days, observation of cell form；On the left-handed and golden nanometer particle thin film of right-handed chirality, cell all grows synapse；Wherein, on the golden nanometer particle thin film of left-handed chirality, cell is slender type, and a two ends grows synapse, and on the golden nanometer particle thin film of right-handed chirality, surrounding all has synapse to grow.
Claims (2)
1. the method that a neuroblastoma is broken up on chiral gold nanoparticulate thin films, it is characterised in that step is:
(1) neuroblastoma cultivation on chiral gold nanoparticulate thin films: left-handed or right-handed chirality golden nanometer particle thin film is placed in the cell culture plate well in 6 holes；With the neuroblastoma cell NG108-15 5min in 5mL trypsinization 25mL Tissue Culture Flask, after pancreatin is outwelled, by 1640 culture medium containing 20% hyclone, cell is blown afloat；Add containing 2 × 10 in being placed with the cell culture plate well of golden nanometer particle thin film of left-handed or right-handed chirality4The culture medium of individual cell/mL, is placed in 37 DEG C of CO2 gas incubator cultivation；Observation of cell form and density after 24h, on the golden nanometer particle thin film of left-handed chirality, cell is strip extended position, on the golden nanometer particle thin film of right-handed chirality, cell is rounded, and on the golden nanometer particle thin film of left-handed chirality, cell density is more than cell density on the golden nanometer particle thin film of right-handed chirality；
(2) neuroblastoma differentiation on chiral gold nanoparticulate thin films: after cell cultivates 24h on the left-handed and golden nanometer particle thin film of right-handed chirality respectively, the culture medium more renewed, add retinoic acid extremely final concentration of 1 μM, after continuing to cultivate 2 days, observation of cell form；On the left-handed and golden nanometer particle thin film of right-handed chirality, cell all grows synapse；Wherein, on the golden nanometer particle thin film of left-handed chirality, cell is slender type, and a two ends grows synapse, and on the golden nanometer particle thin film of right-handed chirality, surrounding all has synapse to grow.
The method that the most according to claim 1, neuroblastoma is broken up on chiral gold nanoparticulate thin films, it is characterised in that: the preparation process of described chiral gold nanoparticulate thin films is as follows:
(1) golden nanometer particle of 20nm particle diameter synthesizes and L/D type cysteine modified:
The synthesis of a, 3-4nm gold nano seed: use sodium borohydride reduction synthesis；Take 120mL ultra-pure water, be sequentially added into 147 μ L mass concentration 1% trisodium citrate and 0.5mL
10mM gold chloride, after being sufficiently mixed, adds the sodium borohydride of the 0.1M that 0.6mL newly prepares, i.e. obtains 3-4nm gold nano seed solution when being stirred vigorously；
The golden nanometer particle synthesis of b, 20nm particle diameter: use seed mediated growth method synthesis；5mL 10mM gold chloride is added in 195mL ultra-pure water, it is heated to boiling, it is simultaneously introduced 1.5mL gold nano seed solution and the trisodium citrate of 800 μ L 1% that step a prepares, continues to boil to color constant, i.e. obtain the solution of gold nanoparticles of 20nm particle diameter；
Synthetic solution of gold nanoparticles 9000rpm is centrifuged 10min, and 25 DEG C of concentrations are resuspended in the ultra-pure water of original solution 1/10 volume, regulate pH to 10 with sodium hydroxide, stand-by；Taking two parts of solution, be separately added into L-type cysteine and D type cysteine extremely final concentration of 100 μ g/mL, be stirred overnight, 9000rpm is centrifuged, and removes cysteine unnecessary in solution, resuspended, obtains the left-handed and solution of gold nanoparticles of right-handed chirality；
(2) preparation of golden nanometer particle thin film and transfer: the solution of gold nanoparticles of the left-handed or right-handed chirality that step (1) gained has been modified L/D type cysteine is poured in small beaker, adds 5mL normal hexane, is then slowly added dropwise dehydrated alcohol；During dropping, gold film can gradually on interface, be transferred on polydimethylsiloxane film by emersion by gold film, obtains having the left-handed or golden nanometer particle thin film of right-handed chirality, dries stand-by.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610592436.3A CN105950559B (en) | 2016-07-26 | 2016-07-26 | A kind of method that neuroblastoma is broken up on chiral gold nanoparticulate thin films |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610592436.3A CN105950559B (en) | 2016-07-26 | 2016-07-26 | A kind of method that neuroblastoma is broken up on chiral gold nanoparticulate thin films |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105950559A true CN105950559A (en) | 2016-09-21 |
| CN105950559B CN105950559B (en) | 2019-05-24 |
Family
ID=56898911
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610592436.3A Active CN105950559B (en) | 2016-07-26 | 2016-07-26 | A kind of method that neuroblastoma is broken up on chiral gold nanoparticulate thin films |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105950559B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109126477A (en) * | 2018-09-13 | 2019-01-04 | 浙江工业大学 | A kind of preparation method of chiral separation film |
| CN112552916A (en) * | 2020-12-29 | 2021-03-26 | 江南大学 | Synthesis method of chiral europium selenide nano particles |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105621352A (en) * | 2015-12-31 | 2016-06-01 | 江南大学 | Preparing method of gold nanometer particle single-layer film with chirality |
-
2016
- 2016-07-26 CN CN201610592436.3A patent/CN105950559B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105621352A (en) * | 2015-12-31 | 2016-06-01 | 江南大学 | Preparing method of gold nanometer particle single-layer film with chirality |
Non-Patent Citations (3)
| Title |
|---|
| ANDREW A.等: "Towards the fabrication of artificial 3D microdevices for neural cell networks", 《BIOMED MICRODEVICES》 * |
| PENGYANG WANG等: "Interaction of gold nanoparticles with proteins and cells", 《SCI. TECHNOL. ADV. MATER.》 * |
| 江龙等: "金纳米颗粒的细胞毒性和促细胞生长作用", 《化学进展》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109126477A (en) * | 2018-09-13 | 2019-01-04 | 浙江工业大学 | A kind of preparation method of chiral separation film |
| CN112552916A (en) * | 2020-12-29 | 2021-03-26 | 江南大学 | Synthesis method of chiral europium selenide nano particles |
| WO2022142747A1 (en) * | 2020-12-29 | 2022-07-07 | 江南大学 | Chiral europium selenide nanoparticle synthesis method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105950559B (en) | 2019-05-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Yu et al. | Hydrogen‐Bonded Organic Framework (HOF)‐Based Single‐Neural Stem Cell Encapsulation and Transplantation to Remodel Impaired Neural Networks | |
| Ma et al. | Ultrasound-activated Au/ZnO-based Trojan nanogenerators for combined targeted electro-stimulation and enhanced catalytic therapy of tumor | |
| CN109602919B (en) | Black phosphorus quantum dot coated by core-shell metal organic framework and preparation method and application thereof | |
| CN110623939B (en) | Preparation method of cantharidin-loaded tumor cell membrane encapsulated tellurium elementary substance nanoparticles | |
| Lang et al. | Integration of TiO2 into the diatom Thalassiosira weissflogii during frustule synthesis | |
| Liu et al. | Blue and cyan fluorescent carbon dots: one-pot synthesis, selective cell imaging and their antiviral activity | |
| CN102559600A (en) | Artificial antigen presenting cell and application thereof in NK (natural killer) cell amplification | |
| Jesion et al. | Graphene and carbon nanocompounds: biofunctionalization and applications in tissue engineering | |
| CN105950559A (en) | Method for neuroblastoma differentiation on chiral gold nanoparticle film | |
| Reding et al. | Manipulate intestinal organoids with niobium carbide nanosheets | |
| CN111407889B (en) | Composite material for simultaneously generating oxygen and active oxygen under near infrared light excitation and preparation method and application thereof | |
| CN105670998B (en) | A kind of method of calcification cancer cell | |
| Park et al. | Silica-capped and gold-decorated silica nanoparticles for enhancing effect of gold nanoparticle-based photothermal therapy | |
| CN106880847A (en) | Multifunctional amorphous ferrum nano material and its preparation method and application | |
| CN113897300B (en) | Bifidobacterium animalis for improving skin barrier function injury and skin sensitivity | |
| CN114887061A (en) | Preparation method and application of photo-thermal gene combination therapy nano system for targeting tumors | |
| CN110642865B (en) | Application of high-charge cationic porphyrin in preparation of PDT nano photosensitizer | |
| CN113769111A (en) | Dual miRNA (micro ribonucleic acid) -triggered universal key-drug delivery system as well as construction method and application thereof | |
| CN114196622B (en) | Application of fullerene and derivative thereof in cell reprogramming | |
| CN113117078A (en) | Novel tumor treatment drug AuNCs @ GTTN and preparation method and application thereof | |
| Xie et al. | Application of Electrically Charged Carbon Nanodots@ glass in DNA and Cell Engineering | |
| Hao et al. | Tetrahedral DNA Nanostructure-modified Gold Nanorod-based Anticancer Nanomaterials for Combined Photothermal Therapy and Chemotherapy | |
| CN114652832B (en) | Organic metal nano particle and preparation method and application thereof | |
| Kong et al. | Nature‐Inspired Thylakoid‐Based Photosynthetic Nanoarchitectures for Biomedical Applications | |
| CN103210855A (en) | Mantle piece treatment liquid for improving seawater pearl quality and treatment method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |