CN106399231A - Application of nanometer graphene oxide to promotion of culture and self-renewal of mouse embryonic stem cells - Google Patents

Application of nanometer graphene oxide to promotion of culture and self-renewal of mouse embryonic stem cells Download PDF

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CN106399231A
CN106399231A CN201610817382.6A CN201610817382A CN106399231A CN 106399231 A CN106399231 A CN 106399231A CN 201610817382 A CN201610817382 A CN 201610817382A CN 106399231 A CN106399231 A CN 106399231A
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graphene oxide
embryonic stem
culture
stem cell
self
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CN106399231B (en
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汪世龙
朱融融
静国欣
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Tongji University
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    • 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
    • C12N5/0603—Embryonic cells ; Embryoid bodies
    • C12N5/0606—Pluripotent embryonic cells, e.g. embryonic stem cells [ES]
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    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/32—Amino acids
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    • C12N2500/00—Specific components of cell culture medium
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    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/44—Thiols, e.g. mercaptoethanol
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    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/20—Cytokines; Chemokines
    • C12N2501/23—Interleukins [IL]
    • C12N2501/235—Leukemia inhibitory factor [LIF]

Abstract

The invention relates to application of nanometer graphene oxide to promotion of culture and self-renewal of mouse embryonic stem cells. Graphene oxide can promote self-renewal of embryonic stem cells and inhibit cell differentiation under the culture condition of no leukemia inhibitory factor (LIF). The surface of graphene oxide has a plurality of functional groups, so graphene oxide has the characteristics of good biological compatibility, low cytotoxicity, etc., and graphene oxide is low in cost, convenient in material synthesis and good in effect and has wide application prospects in research on embryonic stem cells.

Description

Nano graphene oxide is in promoting Mouse Embryonic Stem Cell Culture and self renewal Application
Technical field
The present invention relates to a kind of application in promoting mouse embryo cell culture and self renewal for nano graphene oxide.
Background technology
Nanotechnology is emerging in modern science and a field of high speed development.In the past few years, nanotechnology quilt Efficiently apply in biomedical research, be cell imaging, biological in the delivery of gene or other small molecules and organizational project The preparation of support provides new method.
Embryonic stem cell（ESCs）It is the cell separated from the blastula stage inner cell mass of embryo, because it has simultaneously Unlimited competence for added value and multi-lineage potential and be widely used in biomedical and regenerative medicine research.Up to the present, ESCs is gradually applied to the treatment of clinical disease, such as spinal cord injury, parkinson collective species, myocardial infarction and pulmonary fibrosiss etc.. Therefore, to ESCs propagation or differentiation accuracy controlling be current research key.Research shows, the knot of nanotechnology and stem cell Conjunction may be the culture of stem cell and regulate and control to bring important breakthrough, research before focuses primarily upon separation spike, the medicine of stem cell Thing and the delivery of gene.Nowadays research range gradually spreads over nano material and stem cell self renewal, differentiation and cell is ordered In the regulation and control of fortune.Correlational study shows, nano layered double hydroxides（LDH）There is replacement leukaemia inhibitory factor LIF, maintain The self renewal of mouse embryo stem cell, the effect of Inhibited differentiation.Nano layered double hydroxides are a kind of anionic inorganic pieces Stratified material, because it has good biocompatibility and hypotoxicity, is widely used in biomedical research.
Graphene oxide（Graphene Oxide, GO）It is a kind of sheet material, there are electric conductivity, good biofacies Capacitive and surface modificability, are more and more applied in biological study.Correlational study shows, graphene oxide pair The propagation of multiple stem cell, differentiation have regulating and controlling effect.
Content of the invention
It is an object of the invention to provide a kind of nano graphene oxide is promoting mouse embryo cell culture and self renewal In application.
Application in promoting mouse embryo cell culture and self renewal for the nano graphene oxide proposed by the present invention, tool Body step is as follows：
(1）The mouse embryo stem cell of normal culture is removed by feeder layer cells by differential attachment method, by it according to 20000 Individual/hole is seeded on six orifice plates being covered with 1% gelatin；
（2）By step（1）After the product obtaining adopts complete medium to cultivate 24 hours, remove the LIF factor, in complete medium Middle interpolation nano graphene oxide, beginning nano graphene oxide final concentration maintains 10 μ g/mL-40 μ g/mL, continues culture mice Embryonic stem cell, after interpolation nano graphene oxide after 48 hours, you can show that from form it is able to maintain that mouse embryonic stem The self-renewal capacity of cell；Wherein：Complete medium is by DMEM in high glucose；15% hyclone；0.1mmol/L non-essential amino Acid；2mmol/L L-Glutamine；0.1mmol/L beta -mercaptoethanol；1000U/mL leukaemia inhibitory factor forms.
By alkaline phosphatase staining, Western Blot, quantitative fluorescent PCR (rt-PCR) and immunofluorescence dyeing, Show that graphene oxide has the self renewal power maintaining mouse embryo stem cell and suppresses the ability of its differentiation.
The beneficial effects of the present invention is；The present invention proposes nano graphene oxide（GO）Cultivate field for stem cell New application, removing feeder layer cells and no leukaemia inhibitory factor（LIF）In the presence of, it is able to maintain that mice embryonic The self-renewal capacity of stem cell.Embryonic stem cell easily breaks up during long-term cultivation, in order to maintain embryonic stem cell Self-renewal capacity, the culture of traditional embryonic stem cell need to rely on feeder layer cells and leukocyte inhibitory factor, this Make that the toxigenic capacity of embryonic stem cell is high, incubation is loaded down with trivial details, and the present invention can simplify the culture flow process of embryonic stem cell, Reduce the toxigenic capacity of embryonic stem cell.
Brief description
Fig. 1 is the transmission electron microscope picture of graphene oxide.
Fig. 2 is that the graphene oxide of variable concentrations processes embryonic stem cell respectively 24 hours, after 48 hours, cell produced The MTT testing result of toxicity.Wherein：A () is 24 hours, (b) is 48 hours.
Fig. 3 for variable concentrations graphene oxide process 4 days after mouse embryo stem cell alkaline phosphatase staining.Wherein： (a) be culture fluid in add the LIF factor and（b）Remove the LIF factor as a control group.Final concentration difference after the dilute addition of graphite oxide For (c) 4mg/mL, (d) 8mg/mL, (e) 16mg/mL, (f) 32mg/mL, (g) 40mg/mL.
Fig. 4 detects the expression feelings of mouse embryo stem cell multipotency and self renewal associated protein for Western Blot Condition.
Fig. 5 detects the expression of pluripotency gene for immunofluorescence dyeing.Wherein：（a）、（b）、（c）、（d）Figure is respectively not Mouse embryo stem cell self renewal marker gene Nanog after cultivating 4 days under conditions of adding the lif factor, Oct4, Sox2 With the expression of SSEA-1,（e）、（f）、（g）、（h）Figure adds graphene oxide under conditions of respectively removing the lif factor Mouse embryo stem cell self renewal marker gene Nanog after cultivating 4 days, the expression feelings of Oct4, Sox2 and SSEA-1 Condition.
Specific embodiment
Combine accompanying drawing below by embodiment and further illustrate the present invention.
Embodiment 1：The effect flow operations of the culture of mouse embryo stem cell and graphene oxide
Primary acquisition mouse embryo fibroblasts（MEF）, cultivate to 3-5 generation, the gamma-radiation through 80kV is made after processing 30min For feeder layer cells, use in 1 week.Mouse embryo stem cell adopts complete medium：DMEM in high glucose, 15% hyclone, 0.1 Mmol/L non essential amino acid, 2 mmol/L L-Glutamine, 0.1 mmol/L beta -mercaptoethanol, 1000U/ml0.1 leukemia presses down The factor processed, after normally cultivating 2-3 days, plus trypsin（0.25%）-EDTA（0.02%）Peptic cell, in culture fluid and after pancreatin Cell is placed on the culture dish being coated 0.1%, quiescent culture 30 minutes.After drawing not adherent mESC counting, according to 2 × 104 Individual/hole is seeded on paving coated six orifice plates of 1% gelatin.
After culture 24 hours, remove the LIF factor, the graphene oxide adding size to be 200nm in culture fluid is molten Liquid is so as to final concentration is in 10-40 μ g/mL, the daily culture fluid changing the graphene oxide with same concentrations, continuous processing 4 After it, carry out the detection of mouse embryo stem cell self-renewal capacity index of correlation.
Embodiment 2：Mtt assay detects cell survival rate
MESCs is unicellular through digesting, blowing and beating into, removes through differential attachment method after feeder layer cells, according to 2 × 103Individual/hole close Degree is inoculated on 96 orifice plates being coated gelatin, every hole 200 μ L culture fluid, incubated overnight.Next day, cell culture fluid is replaced by Culture fluid containing graphene oxide, the concentration of graphene oxide is respectively 0,0.5,1,1.5,2,2.5,3 μ g/mL.Change not The cell adding graphene oxide culture fluid is set to blank control group.Experimental group and matched group all set up 5 multiple holes.Respectively in oxygen Graphite alkene processes 24 h, 48 h detect to cell.Prepare 5 mg/mL brominations 3-（4,5- dimethylthiazole -2）-2,5- Diphenyltetrazolium bromide [3-（4,5-dimethylthiazol-2yl）-2,5- diphenylterazolium bromide, MTT] solution, with PBS dilution, pH=7.4, every hole adds 20 μ L.After being incubated 4 h, suck in the hole culture fluid, every hole adds 150 μ L DMSO, horizontal oscillations 10 min.With microplate reader read 570 nm excitation wavelengths under each hole absorbance value, record result and according to Formula calculates cell survival rate：Survival rate %=A570（Experimental group）/A570（Blank control group）×100 %.
Embodiment 3：The impact to mouse embryo stem cell self renewal for the graphene oxide
（1）Alkali phosphatase（ALP）Activity determination.One important biomolecule feature of undifferentiated embryonic stem cell is being capable of table Reach the alkali phosphatase (AP) of high concentration, the differentiation state of embryonic stem cell can be reflected by alkali phosphatase.To by reality Carry out alkaline phosphatase staining after the mESC fixation applying 1 acquisition.As shown in Fig. 2 from form, having removed the mESC of the LIF factor Break up completely, add the mESC of graphene oxide to maintain undifferentiated state, and add the mESC of graphene oxide to be in alkaline phosphatase Enzyme strong positive；Illustrate that graphene oxide can promote the self-renewal capacity of mESC.
（2）Western Blot detects the expression of mouse embryo stem cell multipotency and self renewal associated protein. These albumen of Nanog, Oct4, Sox2, Rex-1 have important regulating and controlling effect to the self renewal of ES cell.Western Blot can detect the expression of these albumen, reflects cell state in which.As shown in figure 4, graphene oxide can be gone up Adjust the expression of this four genes, promote mouse embryo stem cell self-renewal capacity.
（3）Immunofluorescence detects the expression of pluripotency gene on a cellular level.From Rabbit anti-Nanog, Rabbit anti-Oct4, Rabbit anti-Sox2, Mouse anti-SSEA-1 are anti-as 1, Fluorescein Conjugated Goat Anti-Rabbit Secondary Antibody and Fluorescein Conjugated Goat Anti- Mouse Secondary Antibody is anti-as 2, and DAPI carries out nuclear targeting, and fluorescence microscope dyes Situation.As shown in figure 5, compared with matched group, graphene oxide can promote Nanog, Oct4, Sox2 and SSEA-1 in cellular water Flat expression.

Claims (2)

1. application in promoting Mouse Embryonic Stem Cell Culture and self renewal for the nano graphene oxide.
2. the application according to right 1 is it is characterised in that comprise the following steps that：
（1）The mouse embryo stem cell of normal culture is removed by feeder layer cells by differential attachment method, by it according to 20000 Individual/hole is seeded on six orifice plates being covered with 1% gelatin；
（2）By step（1）After the product obtaining adopts complete medium to cultivate 24 hours, remove the LIF factor, in complete medium Middle interpolation nano graphene oxide, beginning nano graphene oxide final concentration maintains 10 μ g/mL-40 μ g/mL, continues culture mice Embryonic stem cell, after interpolation nano graphene oxide after 48 hours, you can show that from form it is able to maintain that mouse embryonic stem The self-renewal capacity of cell；Wherein：Complete medium is by DMEM in high glucose；15% hyclone；0.1mmol/L non-essential amino Acid；2mmol/L L-Glutamine；0.1mmol/L beta -mercaptoethanol；1000U/mL leukaemia inhibitory factor forms.
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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107022517A (en) * 2017-05-25 2017-08-08 句容亿格纳米材料厂 A kind of method for improving glutamine stability in stem cell media
CN107177542A (en) * 2017-05-25 2017-09-19 句容亿格纳米材料厂 A kind of optimization method for promoting culture medium GLN to dissolve
CN109055477A (en) * 2018-08-20 2018-12-21 淮南师范学院 A method of based on determination of immune cells GO biological effect
CN110106141A (en) * 2019-04-24 2019-08-09 朗姿赛尔生物科技（广州）有限公司 A kind of method of autologous stem cells scale
CN111763655A (en) * 2020-09-03 2020-10-13 朗姿赛尔生物科技（广州）有限公司 Method for promoting stem cell expansion
CN111808798A (en) * 2020-09-03 2020-10-23 朗姿赛尔生物科技（广州）有限公司 Culture medium for promoting stem cell expansion
CN111979182A (en) * 2020-09-07 2020-11-24 上海市同济医院 Application of magnesium-iron layered double hydroxide nano material in subculture and cryopreservation of mouse embryonic stem cells
CN112779220A (en) * 2021-04-01 2021-05-11 河南循远生物科技有限公司 Culture medium for neural stem cell amplification

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103820387A (en) * 2014-03-11 2014-05-28 中国科学院上海微***与信息技术研究所 Application of germanium-based graphene in osteogenesis promotion
WO2015157647A2 (en) * 2014-04-10 2015-10-15 Rutgers, The State University Of New Jersey Guiding stem cell differentiation using graphene-nanofiber hybrid scaffolds
CN105062964A (en) * 2015-07-17 2015-11-18 山东大学 Inducing liquid for improving osteogenic differentiation efficiency in stem cells and application of inducing liquid

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103820387A (en) * 2014-03-11 2014-05-28 中国科学院上海微***与信息技术研究所 Application of germanium-based graphene in osteogenesis promotion
WO2015157647A2 (en) * 2014-04-10 2015-10-15 Rutgers, The State University Of New Jersey Guiding stem cell differentiation using graphene-nanofiber hybrid scaffolds
CN105062964A (en) * 2015-07-17 2015-11-18 山东大学 Inducing liquid for improving osteogenic differentiation efficiency in stem cells and application of inducing liquid

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107022517A (en) * 2017-05-25 2017-08-08 句容亿格纳米材料厂 A kind of method for improving glutamine stability in stem cell media
CN107177542A (en) * 2017-05-25 2017-09-19 句容亿格纳米材料厂 A kind of optimization method for promoting culture medium GLN to dissolve
CN109055477A (en) * 2018-08-20 2018-12-21 淮南师范学院 A method of based on determination of immune cells GO biological effect
CN110106141A (en) * 2019-04-24 2019-08-09 朗姿赛尔生物科技（广州）有限公司 A kind of method of autologous stem cells scale
CN110106141B (en) * 2019-04-24 2022-05-20 朗姿赛尔生物科技（广州）有限公司 Method for scaling autologous stem cells
CN111763655A (en) * 2020-09-03 2020-10-13 朗姿赛尔生物科技（广州）有限公司 Method for promoting stem cell expansion
CN111808798A (en) * 2020-09-03 2020-10-23 朗姿赛尔生物科技（广州）有限公司 Culture medium for promoting stem cell expansion
CN111763655B (en) * 2020-09-03 2021-01-08 朗姿赛尔生物科技（广州）有限公司 Method for promoting stem cell expansion
CN111979182A (en) * 2020-09-07 2020-11-24 上海市同济医院 Application of magnesium-iron layered double hydroxide nano material in subculture and cryopreservation of mouse embryonic stem cells
CN111979182B (en) * 2020-09-07 2023-08-15 上海市同济医院 Application of magnesium-iron layered double hydroxide nano material in mouse embryonic stem cell subculture and cryopreservation
CN112779220A (en) * 2021-04-01 2021-05-11 河南循远生物科技有限公司 Culture medium for neural stem cell amplification
CN112779220B (en) * 2021-04-01 2023-10-10 新疆赛尔托马斯生物科技有限公司 Culture medium for neural stem cell expansion

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