CN107988157B - Dendritic cell induction culture medium and application thereof - Google Patents
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Abstract
The invention relates to the technical field of cell culture, in particular to a dendritic cell induction culture medium and application thereof. The culture medium is characterized in that the following components are added into RPMI-1640 culture medium according to the volume of the culture medium: the volume fraction of the fetal calf serum is 8-12%, hrGM-CSF 5-10 ng/ml, IL-45-10 ng/ml, IL-3100-160 ng/ml, SCF 60-120 ng/ml, hrBDNF 60-100 ng/ml and CYM-5442120-200 ng/ml. The culture medium takes hrBDNF and CYM-5442 as key regulatory factors, so that the time for inducing human cord blood mononuclear cells into DC can be shortened to about 10 days, and the obtained DC has excellent biological activity.
Description
Technical Field
The invention relates to the technical field of cell culture, in particular to a dendritic cell induction culture medium and application thereof.
Background
Dendritic Cells (DCs) belong to the leukocyte category of the originating stem cells, are the most important antigen-presenting cells in the initial immune response, and specifically activate T natural cells. The immune function of dendritic cells of tumor patients is incomplete, and the antigen processing function of tumor cells is down-regulated, low-expressed or does not express costimulatory molecules, so that the dendritic cells are the main reasons for immune escape. Dendritic cell therapy is that patient's autologous mononuclear cells are used for in vitro culture induction to generate DC, then corresponding tumor antigens are loaded to prepare dendritic cells loaded with the tumor antigens, and the dendritic cells are injected into a human body to stimulate the proliferation of tumor killing lymphocytes in the human body so as to play a long-term tumor monitoring role and a tumor killing role and achieve the purpose of eliminating tumors. Immunotherapy, which uses antigen-presenting cells-dendritic cells as the center, is one of the main directions of tumor biological therapy.
The acquisition of dendritic cells is a necessary prerequisite for the research thereof, however, the content of dendritic cells and precursors thereof in vivo is very small, and the induction of peripheral blood mononuclear cells in vitro to obtain a sufficient number of dendritic cells for biotherapy has difficulties in specimen source and technology.
The cord blood has rich sources and is not used all the time. A great deal of research in recent years has found that hematopoietic stem cells are present in greater and more primitive quantities per unit volume of cord blood than bone marrow and peripheral blood stem cells. It has been reported that mononuclear cells from peripheral blood and cord blood can be differentiated into functional dendritic cells after coculture with GM-CSF, IL-4, and TNF α. However, the DC in vitro maturation promotion scheme in the prior art has no unified standard, TNF-a factors are mainly adopted at home, but the maturation degree of the DC activated by the method is low, and proinflammatory factor IL-12 cannot be secreted, so that the DC obtained by culture has functional defects and the culture time is long. The development of the clinical application work of the DC vaccine is greatly limited. Therefore, there is a need for further improvements in existing DC cell culture protocols to effectively improve the role of DC vaccines in inducing anti-inflammatory or anti-tumor immune responses.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The first purpose of the invention is to provide a Dendritic Cell (DC) induction culture medium, which takes an S1P1 receptor agonist CYM-5442 and a human recombinant brain-derived neurotrophic factor (hrBDNF) as key regulating factors, adapts to and adjusts the original Dendritic cell induction culture medium, and can be well applied to DC induction maturation and clinical application.
The second purpose of the invention is to provide an application of the dendritic cell induction culture medium in inducing the differentiation of the human umbilical cord blood mononuclear cells to the DC, wherein the dendritic cell induction culture medium can induce the human umbilical cord blood mononuclear cells to differentiate and mature to the DC, the induction culture time is shorter, and the obtained DC can secrete more proinflammatory factor IL-12.
In order to achieve the above purpose of the present invention, the following technical solutions are adopted:
a dendritic cell induction culture medium is prepared by adding the following components in RPMI-1640 culture medium according to the volume of the culture medium:
the volume fraction of the fetal calf serum is 8-12%, hrGM-CSF 5-10 ng/ml, IL-45-10 ng/ml, IL-3100-160 ng/ml, SCF 60-120 ng/ml, hrBDNF 60-100 ng/ml and CYM-5442120-200 ng/ml.
Brain derived neurotrophic factor BDNF (brain derived neurotrophic factor) plays a key role in inflammation and allergic reactions. Previous studies have shown that BDNF and NGF stimulate DC activation in inflammatory responses and promote the release of IL6 or IL10 (Clinical and Experimental Allergy,37, 1701-1708, 2007, o.noga & m.peiser et al); however, BDNF has been rarely studied in inducing DC differentiation and maturation.
CYM-5442 is an agonist of S1P1(Sphingosine-1-phosphate Sphingosine-1 phosphate receptor), and studies show that CYM-5442 can enter DC and regulate DC to exert immune function, and also indicate that the expression level of S1P1 receptor in mature DC is increased, but the research of CYM-5442 in inducing DC differentiation and maturation is not seen in the field.
According to the invention, BDNF and CYM-5442 with appropriate concentrations are added into a dendritic cell induction culture environment, so that the induction maturation speed of dendritic cells can be greatly increased, and the cooperation of the BDNF and the CYM-5442 has a synergistic effect. In the prior art, the hrGM-CSF, IL-4, IL-3 and SCF are all common DC induction medium components, but the IL-4 and IL-3 are used less in combination, and the application finds that the combination of IL-4 and IL-3 with hrBDNF and CYM-5442 has better induction effect.
The invention also relates to application of the dendritic cell induction culture medium in inducing the differentiation of the human umbilical cord blood mononuclear cells to the dendritic cells.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention provides a dendritic cell induction culture medium, which takes hrBDNF and CYM-5442 as key regulating factors, constructs a finished dendritic cell induction culture medium on the basis of the existing dendritic cell induction culture medium, and is particularly suitable for the relevant research and clinical application of the differentiation of human umbilical blood mononuclear cells to dendritic cells;
(2) in order to enhance the synergistic cooperation effect among the components and obtain the dendritic cells with better expression, the invention optimizes and limits the proportion among the components of the dendritic cell induction culture medium;
(3) by using the dendritic cell induction culture medium provided by the invention, the time for inducing the human umbilical cord blood mononuclear cells into DC can be shortened to about 10 days, and the secretion of IL-12p70 and IL-10 is greatly improved compared with the prior art.
Detailed Description
The invention relates to a dendritic cell induction culture medium, which is characterized in that the following components are added into an RPMI-1640 culture medium according to the volume of the culture medium:
the volume fraction of the fetal calf serum is 8-12%, hrGM-CSF 5-10 ng/ml, IL-45-10 ng/ml, IL-3100-160 ng/ml, SCF 60-120 ng/ml, hrBDNF 60-100 ng/ml and CYM-5442120-200 ng/ml.
In order to achieve better culture effect, preferably, the fetal calf serum is special grade fetal calf serum.
In order to enable the dendritic cell induction culture medium to better exert the induction effect and obtain the dendritic cells with more obvious properties, the volume fraction of fetal bovine serum is preferably 9-11%, hrGM-CSF 7-8 ng/ml, IL-47-8 ng/ml, IL-3120-140 ng/ml and SCF 80-100 ng/ml in terms of the volume of the culture medium;
more preferably, the volume fraction of fetal calf serum is 10%, hrGM-CSF7.5ng/ml, IL-47.5ng/ml, IL-3130ng/ml, SCF 90ng/ml, based on the volume of the culture medium.
Preferably, the dendritic cell induction culture medium comprises 70-90 ng/ml of hrBDNF and 78-180 ng/ml of CYM-5442140 based on the volume of the culture medium.
Preferably, the dendritic cell induction culture medium comprises 75-85 ng/ml of hrBDNF and 75-170 ng/ml of CYM-5442150 based on the volume of the culture medium.
Preferably, the dendritic cell induction medium is hrBDNF 80ng/ml and CYM-5442160 ng/ml based on the volume of the medium.
In addition, in order to prevent the cells from being contaminated during the culture process, it is preferable that the dendritic cell induction medium as described above further includes:
penicillin is 90-110U/ml, and streptomycin is 90-110U/ml; more preferably penicillin 100U/ml and streptomycin 100U/ml.
According to one aspect of the invention, the invention also relates to the application of the dendritic cell induction culture medium as described above in inducing the differentiation of the human umbilical cord blood mononuclear cells to the dendritic cells.
Preferably, the time for cell induction culture is 8-12 days, and more preferably 10 days.
Preferably, the human cord blood mononuclear cells are used as described above at 1X 104Density inoculation per well for culture.
Preferably, the culture medium is replaced 1 time every 2 to 3 days in the culture process by using the method.
Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
Examples
Primary reagent
1. Granulocyte-macrophage colony stimulating factor (rh GM-CSF): r & D Systems, Minneapolis, MN, USA.
2. Interleukin-3 (IL-3): kylin Kunpeng biological pharmaceutical Co., Ltd.
3. Interleukin-4 (IL-4): r & D Systems, Minneapolis, MN, USA.
4. Stem cell growth factor (SCF): kylin Kunpeng biological pharmaceutical Co., Ltd.
hrBDNF: PeproTech Inc.
CYM-5442: Sigma-Aldrich.
7. Fetal bovine serum Hyclone.
hrNGF-. beta.Sigma-Aldrich.
EPO: Sigma-Aldrich.
(II) Main instrument
1. Carbon dioxide constant temperature incubator: model 3336, Forma Scienfic, USA
2. Superclean bench: SW-CT-1 type B, Suzhou.
3. A low-temperature refrigerator: model MDF-382E, SANYO, Japan.
(III) Experimental methods
1. Preparing cord blood mononuclear cells.
Preparing cord blood mononuclear cells: when the baby is born, the cord blood is collected by an 18G needle, heparin is used for anticoagulation, and the cord blood is collected each time15-35 ml of blood. Centrifuging the cord blood at 2500 rpm at normal temperature for 10 min, removing the upper plasma layer, diluting with PBS, suspending on Ficoll-Paque lymphocyte separating medium (d is 1.077), centrifuging at 1500 rpm at room temperature for 20 min, collecting cord blood mononuclear cells between separating media, washing with PBS, preparing mononuclear cell suspension, and regulating cell suspension concentration to 5 × 106/ml。
2. Culturing cord blood cells: 1X 106The mononuclear cells of the cord blood are cultured in the bottom area of 25m2In the plastic culture bottle and the culture solution containing the cell factors rh GM-CSF, IL-3, SCF and EPO, the fresh culture solution is replaced and the cell factors are added at intervals of 7 days under the conditions of 37 ℃, 5 percent of carbon dioxide and saturated humidity.
3. Dendritic cell in-vitro induction amplification culture system:
experimental groups: RPMI-16409 ml, fetal bovine serum 1ml, hrGM-CSF7.5ng/ml, IL-47.5ng/ml, IL-3130ng/ml, SCF 90ng/ml, hrBDNF 80ng/ml, CYM-5442160 ng/ml, penicillin 100U/ml, streptomycin 100U/ml.
Control group 1: RPMI-16409 ml, fetal bovine serum 1ml, hrGM-CSF7.5ng/ml, IL-47.5ng/ml, IL-3130ng/ml, SCF 90ng/ml, hrNGF 80ng/ml, CYM-5442160 ng/ml, penicillin 100U/ml, streptomycin 100U/ml.
Control group 2: RPMI-16409 ml, fetal bovine serum 1ml, hrGM-CSF7.5ng/ml, IL-47.5ng/ml, IL-3130ng/ml, SCF 90ng/ml, hrBDNF 240ng/ml, penicillin 100U/ml, streptomycin 100U/ml.
Control group 3: RPMI-16409 ml, fetal bovine serum 1ml, hrGM-CSF7.5ng/ml, IL-3130ng/ml, SCF 90ng/ml, penicillin 100U/ml, streptomycin 100U/ml.
Control group 4: RPMI-16409 ml, fetal bovine serum 1ml, hrGM-CSF7.5ng/ml, EPO0.2IU/ml, IL-3130ng/ml, SCF 90ng/ml, hrBDNF 80ng/ml, CYM-5442160 ng/ml, penicillin 100U/ml, streptomycin 100U/ml.
Blank control group: cord blood mononuclear cells cultured without induction by a dendritic cell in vitro induction culture medium.
Subjecting the mononuclear cells of human cord blood prepared in the above step to 1 × 104Density of holeSeeded in 6-well cell culture plates at 37 ℃ with 5% CO2The culture is carried out.
The differentiation medium was changed 1 time 2 to 3 days, the morphological change of the cells was observed under a microscope during the culture, and the cells were collected 10 days after the culture.
In order to further verify that the culture method of the dendritic cells and the obtained dendritic cells have remarkable beneficial effects, the following experiments are provided for verification.
1. FACS analysis of cell phenotypes
After culturing for 10 days, each group of cells was collected, and the cells were suspended in PBS solution and adjusted to 1X 10 cell concentration5And/ml, adding 100 mu 1 of the mixture into a centrifuge tube, adding fluorescent labeled antibodies (anti-CD 83, CD1a and CD11c) respectively to adjust the final concentration to be 5 mu g/ml, placing the centrifuge tube in a refrigerator at 4 ℃, keeping out of the sun for reaction for 15min, adding a PBS solution to wash for 2 times, taking the fluorescent labeled isotype Ig as a control, detecting the fluorescence intensity of cells by using a computer (FACS calibur), and analyzing by using Cell quest software.
TABLE 1 statistics of cell phenotype FACS
CD1a is a more specific representation of dendritic cells, whereas CD83 is a marker for mature DCs, an integrin highly expressed in CD11c dendritic cells. From the above table, compared with control groups 1-4, the number of CD83+, CD1a + and CD11c + positive cells in the DC induced by the culture medium provided by the invention is significantly increased; and when BDNF (control group 1) or CYM-5442 (control group 2) is lacked in the culture medium, the number of mature DCs is remarkably reduced, which shows that the BDNF and the CYM-5442 can synergistically activate the DCs and have remarkable synergistic effect. Furthermore, control 1 demonstrated that NGF, which functions similarly to BDNF, did not have a synergistic DC-activating effect with CYM-5442. When IL-4 in the medium was replaced with EPO, the number of mature DCs was also reduced, indicating that IL-4 also had some effect on the synergistic effect of BDNF and CYM-5442.
ELISA method for detecting IL-12p70 and IL-10 content of supernatant before and after induction culture
Collecting the supernatant of each group of dendritic cells respectively to carry out ELISA detection, detecting the secretion of IL-12p70 and IL-10 in the supernatant, wherein the detection data are shown in the following table:
TABLE 2 secretion levels (x. + -. s, ng/ml) of IL-12p70 and IL-10
As can be seen from the data in Table 2, the levels of IL-12p70 and IL-10 secreted by dendritic cells in the experimental group are much higher than those of the control group, while IL-12p70 can directly act on CD8+ T cells to proliferate, enhance the cellular immune effect and form long-term memory T lymphocytes, and can secrete high levels of IL-12, which is one of the markers of high bioactivity of dendritic cells: therefore, the biological activity of the dendritic cells obtained by the method is better than that of the dendritic cells obtained by the control group.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (12)
1. A dendritic cell induction medium is characterized in that the following components are added into RPMI-1640 medium according to the volume of the medium:
the volume fraction of the fetal calf serum is 8-12%, 5-10 ng/ml of hrGM-CSF, 45-10 ng/ml of IL-3100-160 ng/ml of IL-SCF, 60-120 ng/ml of HRBDNF, 60-100 ng/ml of CYM-5442120-200 ng/ml.
2. The dendritic cell induction medium of claim 1, wherein the volume fraction of fetal bovine serum is 9% to 11%, hrGM-CSF7 to 8ng/ml, IL-47 to 8ng/ml, IL-3120 to 140ng/ml, SCF80 to 100ng/ml, based on the volume of the medium.
3. The dendritic cell induction medium of claim 1, wherein hrBDNF is 70-90 ng/ml and CYM-5442140-180 ng/ml based on the volume of the medium.
4. The dendritic cell induction medium of claim 3, wherein hrBDNF is 75-85 ng/ml and CYM-5442150-170 ng/ml based on the volume of the medium.
5. Dendritic cell induction medium according to claim 4, characterized in that hrBDNF 80ng/ml, CYM-5442160 ng/ml, calculated on the medium volume.
6. Dendritic cell induction medium according to any of the claims 1-5, characterized in that the medium further comprises, based on the volume of the medium:
penicillin is 90-110U/ml, and streptomycin is 90-110U/ml.
7. The dendritic cell induction medium of claim 6, further comprising penicillin 100U/ml and streptomycin 100U/ml on a medium volume basis.
8. Use of the dendritic cell induction medium of any one of claims 1 to 7 for inducing differentiation of human cord blood mononuclear cells into dendritic cells.
9. The use according to claim 8, wherein the time period for cell induction culture is 8 to 12 days.
10. The use according to claim 9, wherein the time period for cell induction culture is 10 days.
11. The use of claim 8, wherein the human cord blood mononuclear cells are at least 1 x 104Density inoculation per well for culture.
12. The use according to claim 8, wherein the culture medium is changed every 2 to 3 days during the culture.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008001379A3 (en) * | 2006-06-28 | 2009-04-30 | Yeda Res & Dev | Activated myeloid cells for promoting tissue repair and detecting damaged tissue |
| US8247227B2 (en) * | 2006-08-28 | 2012-08-21 | The Cleveland Clinic Foundation | Dendritic cell precursors |
| CN102690783A (en) * | 2012-06-07 | 2012-09-26 | 广西医科大学 | Artocarpus lingnanensis lectin capable of inducing dentritic cells to mature and proliferate in vitro |
| CN105647867A (en) * | 2016-02-28 | 2016-06-08 | 深圳爱生再生医学科技有限公司 | Method for inducing dendritic cells to be mature and dendritic cells |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008001379A3 (en) * | 2006-06-28 | 2009-04-30 | Yeda Res & Dev | Activated myeloid cells for promoting tissue repair and detecting damaged tissue |
| US8247227B2 (en) * | 2006-08-28 | 2012-08-21 | The Cleveland Clinic Foundation | Dendritic cell precursors |
| CN102690783A (en) * | 2012-06-07 | 2012-09-26 | 广西医科大学 | Artocarpus lingnanensis lectin capable of inducing dentritic cells to mature and proliferate in vitro |
| CN105647867A (en) * | 2016-02-28 | 2016-06-08 | 深圳爱生再生医学科技有限公司 | Method for inducing dendritic cells to be mature and dendritic cells |
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