CN110885781A - Human early placenta development model establishing method based on organ chip - Google Patents
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Abstract
The invention discloses a method for establishing a human early placenta development model based on an organ chip, which mainly comprises the following steps: the invention utilizes the self-assembly property of hPSC to form a three-dimensional cavity structure with near physiology in a three-dimensional matrix material, and the cavities also show the efficient differentiation characteristic of trophoblasts. The method has the characteristics of simplicity, easiness in operation, strong repeatability and the like. The establishment of the model can provide a platform for exploring the research field of human placentation and embryogenesis in the cell level and molecular mechanism, and has great application value in the epigenetic research in the placenta development process.
Description
Technical Field
The invention relates to the technical field of placenta developmental research, tissue bionics and cell biology research, in particular to a human early placenta development model establishing method based on an organ chip.
Background
The placenta is an important organ connecting the mother and fetus, and is also an important organ for exchanging substances between the fetus and the mother. The fetus develops in the uterus and relies on the placenta to obtain nutrients from the mother to maintain a normal pregnancy. More and more studies have shown that human childhood and even adult diseases are related to the growth and development of the placenta. After the initiation of the human placenta program in the united states, obstetricians in china have also become increasingly concerned with the study of this vital organ of gestation. The research content relates to the fields of placenta structure, infection and inflammation, genomics, placenta function and the like. Therefore, the method has great application value in exploring the occurrence of human placenta at a cellular level or a molecular level.
The placenta is the first differentiated extra-embryonic tissue, which originates from the outer layer of the blastocyst (5 days after fertilization) and is called the trophoblast. These cells are precursors to trophoblasts, which surround a fluid-filled cavity to protect embryonic cells. After implantation of human embryos in the mother's uterus, they have a special capacity to attach to and invade the uterine wall. Despite the fundamental clinical significance of placental development, early placental development in humans remains a mystery due to the lack of studies on the stage of implantation of human embryos and the significant differences in placental development between humans and other commonly used animal and cell models. Even with recent in vitro research advances, including human in vitro embryo culture, or in the study of early human embryogenesis, the development of the human placenta is unclear.
Human pluripotent stem cells (hPSCs), which resemble embryonic cells in their self-renewing ground state, have been successfully used in studies of mouse embryonic development at the embryo implantation stage. Although valuable experience gained from studies of vertebrate development has helped us to generate human embryos in vitro, the suitability of human hPSCs in modelling early development has not been determined. The induction of hPSCs under traditional 2D conditions can show different trophoblast cell subtypes in different cell growth periods, but the structure and the physiological condition are quite different, and the simulation of the development of the early placenta in a three-dimensional matrix in vitro has great advantages in terms of trophoblast cell differentiation efficiency and function expression besides a near-physiological three-dimensional structure, so that the capability of hPSCs in simulating the formation of the early human placenta in vitro is extremely valuable for research.
Disclosure of Invention
The invention aims to provide a method for establishing a human early placenta development model based on an organ chip, which is used for researching the development of placenta in a 3D matrix, wherein the model established by the method has a three-dimensional structure close to physiology, and compared with a traditional two-dimensional monolayer, cavities also show efficient differentiation of trophoblasts. The model can provide a platform for exploring the research of human placentation and embryogenesis in the cellular level and molecular mechanism.
The invention relates to a method for establishing a human early placenta development model based on an organ chip, which uses a three-dimensional matrix material, wherein the three-dimensional matrix material is a commercially available artificial basement membrane (Matrigel), and is carried out according to the following steps:
(1) modification of pore plate and inoculation culture of human pluripotent stem cells (hPSC)
The 6-well plate is modified by an artificial basement membrane (Matrigel) and placed in a 4-degree refrigerator, then placed in a 37-degree incubator for 20-50min, digested by cell digestive fluid at room temperature, cell clusters are scraped by a cell scraper, centrifugation is carried out for 600r1-3min, cells are resuspended by mTeSR1 culture medium containing Y27632 to enable the cells to be small clusters and then inoculated in the modified 6-well plate, and the culture fluid is replaced by mTeSR1 culture medium without Y27632 after the cells are cultured for 1-4h at 37 ℃.
The dilution multiple range of the artificial substrate membrane used for modifying the 6-hole plate is 1: 500-1: 1000, 1.5ml of each well was placed in a 4 ℃ freezer for a period ranging from 4 hours to 7 days, and the medium contained Y27632 at a concentration ranging from 1 to 100 mM.
(2) Cells were digested and seeded in a three-dimensional matrix
And (3) carrying out warm digestion by using a digestive juice after the hPSC cells grow to 70-90%, scraping cell masses by using a cell scraper, centrifuging for 1-3min, sucking supernatant fluid, diluting matrigel to a certain proportion by using a culture medium, then carrying out heavy suspension on the cells to obtain a proper inoculation density, inoculating, placing in a 37-degree culture box for 20-40min to solidify the matrigel in which the cells are suspended, and then obtaining the culture medium containing the BMP4 for inducing hPSC differentiation.
The matrigel dilution is specifically a culture medium: matrigel volume ratio of 0: 1-3:1, cell seeding density of 1X 104-1×1010cells/ml, medium containing BMP4 for inducing hPSC differentiation, was: 96% by volume of DMED/F12, 1% by volume of a double antibody, 1% by volume of glutamine, 1% by volume of an unnecessary amino acid (MEMNEAA), 1% by volume of ITS, 100ng/ml of heparin and 10ng/ml of BMP4, wherein 1g of BSA is added into every 50ml of the prepared culture medium, and the culture medium containing BMP4 and inducing hPSC differentiation is obtained after dissolution.
(3) Establishment of human early placenta development model
And (3) replacing the culture medium in the step (2) every day, wherein the culture medium is the culture medium for inducing the hPSC differentiation in the step (2), and the time range of model establishment is 4-12 days.
The invention provides an application of a human early placenta development model based on an organ chip, which can be used for the characterization of cell morphology, including the characterization of cell types such as differentiation condition of hPSC cells to trophoblast cells, fused trophoblast cells, invasive trophoblast cells and the like, and the characterization of hPSC induced differentiation under 2D conditions and the like.
The invention utilizes the self-assembly property of hPSC to form a three-dimensional cavity structure with near physiology in a three-dimensional matrix material, and the cavities also show the efficient differentiation characteristic of trophoblasts. The method has the characteristics of simplicity, easiness in operation, strong repeatability and the like. The establishment of the model can provide a platform for exploring the research field of human placentation and embryogenesis in the cell level and molecular mechanism, and has great application value in the epigenetic research in the placenta development process.
Drawings
FIG. 1 is a schematic diagram of the method for establishing a human early placenta development model based on an organ chip.
FIG. 2 is a bright field trace of cells seeded in a three-dimensional matrix according to example 1 of the invention, including the bright field traces of D0, D1, D2, D4, D6, bar: 50 um.
FIG. 3 is a PCR characterization of hPSC cell differentiation into trophoblast, confluent trophoblast, invasive trophoblast, etc. in example 1 of the present invention.
FIG. 4 is a schematic diagram of hPSC-induced differentiation under conventional 2D conditions.
FIG. 5 is a graph of bright field traces of cells induced under 2D conditions in example 2 of the invention, including the bright field plots of D0, D2, D4, bar: 25 um.
FIG. 6 is a graph showing the results of PCR induced under 2D and 3D conditions in the cells of example 2 of the present invention, in which a is the expression of trophoblast-associated genes (CDX2, OCT4, CK7, GATA3), b is the expression of fused trophoblast-associated gene (CGB), and c is the expression of invasive trophoblast-associated gene (HLA-G).
Detailed Description
The following examples further illustrate the invention but are not intended to limit the invention thereto.
Example 1
Establishment of human early placenta development model by using hPSC self-assembly idiosome
The present invention uses a three-dimensional matrix material, which is a commercially available artificial basement membrane (Matrigel) (ref: 354277, Corning, Inc.).
The invention relates to a method for establishing a human early placenta development model based on an organ chip, which adopts the three-dimensional matrix material and specifically comprises the following steps:
(1) modification of pore plate and inoculation culture of human pluripotent stem cells (hPSC)
The 6-well plate is modified by an artificial basement membrane (Matrigel) and placed in a 4-degree refrigerator, then placed in a 37-degree incubator for 20min, digested by cell digestive fluid at room temperature, cell clusters are scraped by a cell scraper, centrifuged for 600r2min, cells are suspended by mTeSR1 culture medium containing Y27632 to be small clusters and then inoculated in the modified 6-well plate, and the culture fluid is replaced by mTeSR1 culture medium containing no Y27632 after being cultured for 1h at 37 ℃.
The dilution multiple range of the artificial substrate membrane used for modifying the 6-hole plate is 1: 1000, 1.5ml of each well was placed in a 4 ℃ freezer for 7 days, and the medium contained Y27632 at a concentration of 10 mM.
(2) Cells were digested and seeded in a three-dimensional matrix
After the hPSC cells grow to 90%, the hPSC cells are subjected to warm digestion by a digestive juice chamber, cell clusters are scraped by a cell scraper, centrifugation is carried out for 600r2min, supernatant fluid is sucked away, matrigel is diluted to a certain proportion by a culture medium, the cells are resuspended to obtain a proper inoculation density and inoculated, the matrigel with the suspended cells is placed in a 37-DEG culture box for 20min to solidify, and then the culture medium containing BMP4 for inducing hPSC differentiation is replaced.
The matrigel dilution is specifically a culture medium: matrigel volume ratio of 0: 1, cell inoculationDensity of 1X 106cells/ml, medium for inducing hPSC differentiation containing BMP4, DMED/F12 with the volume concentration of 96%, diabody with the volume concentration of 1%, glutamine with the volume concentration of 1%, nonessential amino acid (MEMNAAA) with the volume concentration of 1%, ITS with the volume concentration of 1%, heparin with the volume concentration of 100ng/ml and BMP4 with the volume concentration of 1g BSA are added into every 50ml of the prepared medium, the medium for inducing hPSC differentiation containing BMP4 is obtained after dissolution, and the schematic diagram of the method of the placentation model established according to the above steps is shown in figure 1.
(3) Establishment of human early placenta development model
And (3) replacing the culture medium in the step (2) every day, wherein the culture medium is the culture medium containing the BMP4 and inducing the hPSC differentiation in the step (2), the model establishing time is 6 days, a bright field tracing diagram of cells in the three-dimensional matrix is shown in figure 2, and the PCR result characterization of the hPSC differentiation condition from the hPSC cells to the trophoblast cells, the fusion type trophoblast cells, the invasion type trophoblast cells and other cell types in the three-dimensional matrix is shown in figure 3.
Example 2
Characterization of difference of 2D and 3D matrixes in human early placenta development model on cell morphology and function based on organ chip
The same artificial basement membrane and cell seeding density as in example 1 were used to create a model of human placental development. Here, comparing the difference between the case of hPSC-induced differentiation under 2D conditions and the case of 3D conditions in case 1, it is fully demonstrated that the model constructed in the three-dimensional matrix of the present invention has a near-physiological three-dimensional structure, and that these cavities also show efficient differentiation of trophoblasts compared to the conventional two-dimensional monolayer.
The process of inducing hPSC differentiation under 2D conditions is as follows:
(1) modification of pore plate and inoculation culture of human pluripotent stem cells (hPSC)
The 6-well plate is modified by an artificial basement membrane (Matrigel) and placed in a 4-degree refrigerator, then placed in a 37-degree incubator for 50min, digested by cell digestive fluid at room temperature, cell clusters are scraped by a cell scraper, centrifuged for 600r 3min, cells are suspended by mTeSR1 culture medium containing Y27632 to be small clusters and then inoculated in the modified 6-well plate, and the culture fluid is replaced by mTeSR1 culture medium containing no Y27632 after being cultured for 4h at 37 ℃.
The dilution multiple range of the artificial substrate membrane used for modifying the 6-hole plate is 1: 500, 1.5ml of each well was placed in a 4 ℃ freezer for 4 hours, and the medium contained Y27632 at a concentration of 10 mM.
(2) Induction of hPSC differentiation under 2D conditions
hPSC cells were grown to about 90% and then treated with induction medium without BMP4 for 2 days, with daily change.
The medium for inducing hPSC differentiation without BMP4 comprises the following components: DMED/F12 with a volume concentration of 96%, diabody with a volume concentration of 1%, glutamine with a volume concentration of 1%, non-essential amino acid (MEMNEAA) with a volume concentration of 1%, ITS with a volume concentration of 1%, heparin with a volume concentration of 100ng/ml, BMP4 with a volume concentration of 10ng/ml, and 1g bsa is added to each 50ml of the prepared culture medium, and the culture medium containing BMP4 for inducing hPSC differentiation is obtained after dissolution, and then the culture medium containing BMP4 is replaced for treatment for 6 days, so as to establish a schematic diagram of hPSC induced differentiation under the conventional 2D conditions as shown in fig. 4, and a bright field tracing diagram of hPSC differentiation under the conventional 2D conditions as shown in fig. 5.
The invention relates to a method for establishing an organ chip-based human early placenta development model, which is characterized in that 2D and 3D matrixes represent the difference between cell morphology and function and are characterized in that: the characterization of the cell types including differentiation from hPSC cells to trophoblast cells, confluent trophoblast cells, invasive trophoblast cells and the like is obviously different from the characterization of hPSC induced differentiation under 2D conditions, and the PCR result is shown in figure 6, which shows that hPSC shows high-efficiency differentiation of trophoblast cells in a three-dimensional matrix compared with the traditional two-dimensional monolayer. The model may provide a platform for exploring the cellular and molecular mechanisms of human placegenesis and embryogenesis.
Example 3
Establishment of external model anthropomorphic early placenta development model by using hPSC self-assembly idiosome
The present invention uses a three-dimensional matrix material, which is a commercially available artificial basement membrane (Matrigel) (ref: 354277, Corning, Inc.).
The invention relates to a method for establishing an in-vitro simulated human early placenta development model, which adopts the three-dimensional matrix material and specifically comprises the following steps:
(1) modification of pore plate and inoculation culture of human pluripotent stem cells (hPSC)
The 6-well plate is modified by an artificial basement membrane (Matrigel) and placed in a 4-degree refrigerator, then placed in a 37-degree incubator for 40min, digested by cell digestive fluid at room temperature, cell clusters are scraped by a cell scraper, centrifuged for 600r1min, cells are suspended by mTeSR1 culture medium containing Y27632 to be small clusters and then inoculated in the modified 6-well plate, and the culture fluid is replaced by mTeSR1 culture medium containing no Y27632 after being cultured for 3h at 37 ℃.
The dilution multiple range of the artificial substrate membrane used for modifying the 6-hole plate is 1: 800, 1.5ml of the dilution per well was placed in a 4 ℃ freezer for 3 days, and the medium contained Y27632 at a concentration of 5 mM.
(2) Cells were digested and seeded in a three-dimensional matrix
After the hPSC cells grow to 70%, the cells are digested in a digestion solution at a warm temperature, cell clusters are scraped by a cell scraper, centrifugation is carried out for 600r1min, supernatant fluid is sucked away, matrigel is diluted to a certain proportion by a culture medium, the cells are resuspended to obtain a proper inoculation density and inoculated, the matrigel with the suspended cells is placed in a 37-DEG culture box for 20min to solidify, and then the culture medium containing BMP4 for inducing hPSC differentiation is replaced.
The matrigel dilution is specifically a culture medium: the volume ratio of matrigel is 3:1, cell seeding density of 1X 108cells/ml, medium containing BMP4 for inducing hPSC differentiation, was: DMED/F12 with a volume concentration of 96%, diabody with a volume concentration of 1%, glutamine with a volume concentration of 1%, non-essential amino acid (MEMNEAA) with a volume concentration of 1%, ITS with a volume concentration of 1%, heparin with a volume concentration of 100ng/ml, BMP4 with a volume concentration of 10ng/ml, and 1g bsa is added to each 50ml of the prepared culture medium, and the culture medium containing BMP4 for inducing hPSC differentiation is obtained after dissolution, and a schematic diagram of the method of the placentation model established according to the above steps is shown in fig. 1.
(3) Establishment of human early placenta development model
And (3) replacing the culture medium in the step (2) every day, wherein the culture medium is the culture medium containing the BMP4 and inducing the hPSC differentiation in the step (2), the model building time is 8 days, the bright field tracing graph of the cells in the three-dimensional matrix is similar to the result in the figure 2, and the PCR result characterization of the hPSC differentiation condition from the hPSC cells to the trophoblast cells, the fusion type trophoblast cells, the invasion type trophoblast cells and other cell types in the three-dimensional matrix is similar to the figure 3 and is not described too much here.
Claims (6)
1. A human early placenta development model building method based on organ chips is characterized in that: a three-dimensional matrix material, namely an artificial matrix membrane Matrigel, is adopted, and the method comprises the following steps:
(1) modification of orifice plate and inoculation culture of human pluripotent stem cells
Modifying a 6-hole plate by using an artificial basement membrane, placing the modified 6-hole plate in a 4-degree refrigerator for 4 hours to 7 days, then placing the modified 6-hole plate in a 37-degree incubator for 20 to 50 minutes, and carrying out warm digestion by using a cell digestive juice chamber; scraping off cell mass with cell scraper, centrifuging at 600r for 1-3min, re-suspending cells with mTeSR1 culture medium containing Y27632 to make cells into small mass, inoculating to modified 6-well plate, culturing at 37 deg.C for 1-4 hr, and culturing with mTeSR1 culture medium containing no Y27632;
(2) cells were digested and seeded in a three-dimensional matrix
Digesting the hPSC cells at a warm temperature by using a digestive juice after the hPSC cells grow to 70-90%, scraping cell masses by using a cell scraper, centrifuging for 600r1-3min, sucking supernatant, diluting matrigel to a certain proportion by using a culture medium containing BMP4 and inducing hPSC differentiation, then re-suspending the cells to obtain a proper inoculation density and inoculating, placing the matrigel suspended with the cells in a 37-degree culture box for 20-40min to solidify the matrigel, and then still using the culture medium containing BMP4 and inducing the hPSC differentiation;
(3) establishment of human early placenta development model
And (3) replacing the culture medium in the step (2) every day, wherein the culture medium is the culture medium containing the BMP4 for inducing the hPSC differentiation in the step (2), and establishing the model for several days.
2. The organ chip-based hypoxia simulation method of human early placental development microenvironment according to claim 1, wherein the artificial substrate membrane modified 6-well plate of step (1) is specifically: diluting the artificial basement membrane by 500 times and 1000 times, placing 1.5ml of diluent in each hole in a 4-DEG refrigerator for 4 hours to 7 days;
in the mTeSR1 medium containing Y27632, the concentration of Y27632 ranges from 1 to 100 mM.
3. The organ chip-based hypoxia mimicking microenvironment for early placental development of human according to claim 1, wherein the matrigel dilution of step (2) is specifically culture medium: matrigel volume ratio of 0-3:1, cell seeding density of 1 × 104-1×1010cells/ml。
4. The organ chip-based hypoxia mimicking microenvironment for early placental development of human according to claim 1, wherein the culture medium for inducing differentiation of hpscs comprising BMP4 in step (2) comprises: 96% by volume of DMED/F12, 1% by volume of a double antibody, 1% by volume of glutamine, 1% by volume of an unnecessary amino acid (MEMNEAA), 1% by volume of ITS, 100ng/ml of heparin and 10ng/ml of BMP4, wherein 1g of BSA is added into every 50ml of the prepared culture medium, and the culture medium containing BMP4 and inducing hPSC differentiation is obtained after dissolution.
5. The method for modeling human early placental development based on organ chips according to claim 1, wherein said modeling time in step (3) is in the range of 4-12 days.
6. Use of an organ chip based model of early placental development in humans, as established in claim 1, wherein: the model can be used for characterization of cell morphology, including characterization of cell types such as differentiation condition of hPSC cells to trophoblast cells, fused trophoblast cells, invasive trophoblast cells and the like, and characterization of hPSC induced differentiation under 2D conditions.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6630349B1 (en) * | 1998-09-23 | 2003-10-07 | Mount Sinai Hospital | Trophoblast cell preparations |
| US6696065B1 (en) * | 1995-05-04 | 2004-02-24 | Aventis Pastuer Limited | Acellular pertussis vaccines and methods of preparation thereof |
| US20080213387A1 (en) * | 2005-04-16 | 2008-09-04 | Axordia Limited | Cytotrophoblast Stem Cell |
| CN101711890A (en) * | 2009-01-15 | 2010-05-26 | 中国人民解放军军事医学科学院基础医学研究所 | Extracellular matrix gel model used for researching development and differentiation of embryonic stem cells |
| CN101711891A (en) * | 2009-01-15 | 2010-05-26 | 中国人民解放军军事医学科学院基础医学研究所 | Artificial uterus three-dimensional gel model used for researching development and differentiation of embryonic stem cells |
-
2018
- 2018-09-07 CN CN201811041005.3A patent/CN110885781B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6696065B1 (en) * | 1995-05-04 | 2004-02-24 | Aventis Pastuer Limited | Acellular pertussis vaccines and methods of preparation thereof |
| US6630349B1 (en) * | 1998-09-23 | 2003-10-07 | Mount Sinai Hospital | Trophoblast cell preparations |
| US20080213387A1 (en) * | 2005-04-16 | 2008-09-04 | Axordia Limited | Cytotrophoblast Stem Cell |
| CN101711890A (en) * | 2009-01-15 | 2010-05-26 | 中国人民解放军军事医学科学院基础医学研究所 | Extracellular matrix gel model used for researching development and differentiation of embryonic stem cells |
| CN101711891A (en) * | 2009-01-15 | 2010-05-26 | 中国人民解放军军事医学科学院基础医学研究所 | Artificial uterus three-dimensional gel model used for researching development and differentiation of embryonic stem cells |
Non-Patent Citations (2)
| Title |
|---|
| B.P. TELUGU 等: ""Comparison of extravillous trophoblast cells derived from human embryonic stem cells and from first trimester human placentas"", 《PLACENTA》 * |
| 杨艳艳等: "人胚胎干细胞向滋养层分化的研究进展", 《中国科学:生命科学》 * |
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