CN117264868A - Cell microsphere and preparation method thereof - Google Patents
Cell microsphere and preparation method thereof Download PDFInfo
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- CN117264868A CN117264868A CN202311555074.7A CN202311555074A CN117264868A CN 117264868 A CN117264868 A CN 117264868A CN 202311555074 A CN202311555074 A CN 202311555074A CN 117264868 A CN117264868 A CN 117264868A
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- 239000004005 microsphere Substances 0.000 title claims abstract description 69
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 229910052588 hydroxylapatite Inorganic materials 0.000 claims abstract description 43
- 239000002070 nanowire Substances 0.000 claims abstract description 43
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 claims abstract description 43
- 239000000725 suspension Substances 0.000 claims abstract description 32
- 239000006285 cell suspension Substances 0.000 claims abstract description 17
- 238000001179 sorption measurement Methods 0.000 claims abstract description 7
- 230000001954 sterilising effect Effects 0.000 claims abstract description 5
- 238000002156 mixing Methods 0.000 claims abstract description 4
- 210000004027 cell Anatomy 0.000 claims description 93
- 239000001963 growth medium Substances 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 10
- 239000000843 powder Substances 0.000 claims description 9
- 238000011534 incubation Methods 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 4
- 102000004142 Trypsin Human genes 0.000 claims description 3
- 108090000631 Trypsin Proteins 0.000 claims description 3
- 230000001464 adherent effect Effects 0.000 claims description 3
- 238000010009 beating Methods 0.000 claims description 3
- 238000007664 blowing Methods 0.000 claims description 3
- 230000029087 digestion Effects 0.000 claims description 3
- 210000000963 osteoblast Anatomy 0.000 claims description 3
- 239000002243 precursor Substances 0.000 claims description 3
- 238000002791 soaking Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 239000012588 trypsin Substances 0.000 claims description 3
- 210000001185 bone marrow Anatomy 0.000 claims description 2
- 210000001671 embryonic stem cell Anatomy 0.000 claims description 2
- 230000001678 irradiating effect Effects 0.000 claims description 2
- 210000002901 mesenchymal stem cell Anatomy 0.000 claims description 2
- 238000010146 3D printing Methods 0.000 abstract description 6
- 210000001519 tissue Anatomy 0.000 abstract description 6
- 239000011664 nicotinic acid Substances 0.000 abstract description 5
- 210000002220 organoid Anatomy 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 238000012258 culturing Methods 0.000 abstract description 2
- 230000004069 differentiation Effects 0.000 abstract description 2
- 230000000052 comparative effect Effects 0.000 description 21
- 230000001640 apoptogenic effect Effects 0.000 description 12
- 102000004264 Osteopontin Human genes 0.000 description 10
- 108010081689 Osteopontin Proteins 0.000 description 10
- 238000010186 staining Methods 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000002073 fluorescence micrograph Methods 0.000 description 3
- 230000017074 necrotic cell death Effects 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 238000012604 3D cell culture Methods 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012605 2D cell culture Methods 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 230000003436 cytoskeletal effect Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- HJUFTIJOISQSKQ-UHFFFAOYSA-N fenoxycarb Chemical compound C1=CC(OCCNC(=O)OCC)=CC=C1OC1=CC=CC=C1 HJUFTIJOISQSKQ-UHFFFAOYSA-N 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 238000012835 hanging drop method Methods 0.000 description 1
- 238000001027 hydrothermal synthesis Methods 0.000 description 1
- 238000003125 immunofluorescent labeling Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000008611 intercellular interaction Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000004264 monolayer culture Methods 0.000 description 1
- 230000011164 ossification Effects 0.000 description 1
- 230000004072 osteoblast differentiation Effects 0.000 description 1
- 230000009818 osteogenic differentiation Effects 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
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- 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
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- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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Abstract
The invention discloses a cell microsphere and a preparation method thereof, and relates to the technical field of biology. The preparation method of the cell microsphere comprises the following steps: sterilizing the hydroxyapatite nanowire to prepare a hydroxyapatite nanowire suspension; preparing a cell suspension; mixing the hydroxyapatite nanowire suspension with the cell suspension to obtain a mixed suspension; the mixed suspension is inoculated into a low adsorption 96-well plate, and cells in the mixed suspension and the hydroxyapatite nanowires are incubated to form the cell microspheres. The cell microsphere provided by the invention overcomes the problems of complex preparation process and low cell activity of the cell microsphere in the prior art, and can effectively obtain the cell microsphere by co-culturing the hydroxyapatite nanowire and the cell suspension, ensure the activity of the cell microsphere and enhance the differentiation capacity of the cell microsphere. The cell microsphere can also be used for preparing organoids or for biological 3D printing to build large-size high-cell-density bionic tissues.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a cell microsphere and a preparation method thereof.
Background
In tissue engineering and regenerative medicine, traditional 2D cell culture techniques can support cell attachment, diffusion and growth, but in general such techniques cannot create 3D structures related to human natural histology. It has been shown that 2D culture techniques flatten cell morphology, resulting in changes in cytoskeletal structure and nuclear shape, and changes in gene expression and protein synthesis. Furthermore, the high sensitivity of cells in a 2D culture environment to drug reactions may produce misleading bioassays. However, the 3D cell culture model is more advantageous than the monodisperse cell or monolayer cell culture, the sphere culture is a method which is commonly used in the 3D cell culture technology, and the sphere is considered as an ideal model for researching in vitro human development and diseases due to the high cell density and heterogeneity, and meanwhile, the cell microsphere with good biological performance is expected to construct large-size high-cell-density bionic tissues/organoids by adopting a biological 3D printing technology.
3D cell spheres have been produced by a number of processes including hanging drop methods, low adsorption surfaces or microfluidic systems, and sound field controlled formation cultures. In all of the methods listed above, the cell microspheres first self-aggregate to form spheres by some degree of cell-cell interaction, followed by self-secretion of ECM. The dense cell-to-cell attachment results in spatial gradients in oxygen concentration, vascularization, and up-regulation of various growth factors required for cell growth, whereas in general oxygen, nutrients, cytokines, etc. cannot be transported to the sphere core, resulting in cell necrosis in the microsphere core region, making current cell sphere applications challenging. Therefore, how to prepare a composite cell microsphere with good biological characteristics and viability is a technical problem to be solved in the field.
Disclosure of Invention
The invention aims to provide a cell microsphere and a preparation method thereof, which are used for solving the problems in the background technology.
In order to achieve the above object, the present invention provides a method for preparing a cell microsphere, comprising the steps of:
(1) Sterilizing the hydroxyapatite nanowire: soaking the prepared hydroxyapatite nanowire powder in ethanol, irradiating with ultraviolet rays, washing and centrifuging for three times by using PBS after the irradiation is finished, and washing and centrifuging once by using a complete culture medium to obtain sterilized hydroxyapatite nanowire powder;
(2) Taking the sterilized hydroxyapatite nanowire powder in the step (1), and adding a complete culture medium to prepare a hydroxyapatite nanowire suspension;
(3) Cell suspension preparation: the adherent cells in the culture dish are subjected to trypsin digestion and then are blown and centrifuged, supernatant is discarded, and a complete culture medium is added for blowing and beating to obtain a cell suspension;
(4) Mixing the suspension prepared by the sterilized hydroxyapatite nanowire in the step (2) with the cell suspension prepared in the step (3) in proportion to obtain a mixed suspension;
(5) Inoculating the mixed suspension in the step (4) into a low-adsorption 96-well plate, adding the mixed suspension into each well, and incubating the cells and the hydroxyapatite nanowires in the mixed suspension to form the cell microspheres.
Preferably, in the step (1), the diameter of the hydroxyapatite nanowire is 30-50 nm, more preferably 30-40 nm, and the length thereof is 10-25 μm, more preferably 10-15 μm.
Preferably, the concentration of ethanol in the step (1) is 95-99.5%, more preferably 99.5%.
Preferably, the duration of the ultraviolet irradiation in the step (1) is 3-5 hours, more preferably 5 hours; the centrifugal revolution is 1000 to 1500r, more preferably 1500r, and the centrifugal time is 3 to 5min, more preferably 5min.
Preferably, the concentration of the hydroxyapatite nanowire suspension in the step (2) is 0.01-0.08 mug/mug, more preferably 0.02-0.06 mug/mug.
Preferably, the concentration of the cell suspension in step (3) is 4X 10 4 ~5×10 4 And more preferably 4X 10 per mL 4 The cell type is one of bone marrow mesenchymal stem cells, osteoblast precursor cells and embryonic stem cells per mL, more preferably osteoblast precursor cells.
Preferably, the volume ratio of the cell suspension to the hydroxyapatite nanowire suspension in the step (4) is 1:1-1.5, more preferably 1:1.
preferably, the amount of the mixed suspension added to each well of the low adsorption 96-well plate in the step (5) is 150-200. Mu.L, more preferably 200. Mu.L, the incubation temperature is 35-37 ℃, more preferably 37 ℃, and the CO is 2 The concentration is 4.5-5%, more preferably 5%, the incubation and standing time is 24-48 h, more preferably 48h, the culture medium is replaced once in 2-3 days, more preferably once in 2 days, and the volume ratio of the replaced culture medium is 20-50%, more preferably 35-50%.
The invention also provides the cell microsphere obtained by the preparation method.
The invention also provides application of the cell microsphere in preparing organoids and biological 3D printing and constructing large-size high-cell-density bionic tissues.
The cell microsphere and the preparation method thereof have the advantages and positive effects that:
1. the cell microspheres prepared by the low-adhesion pore plate with surface hydrophobicity can ensure uniform size and material-to-cell ratio, and provide a stable material supply for subsequent biological 3D printing.
2. The hydroxyapatite nanowire prepared by the hydrothermal synthesis is in a bundled array, so that more stable contact and more contact sites can be provided for cell formation.
3. The hydroxyapatite nanowire can be combined with a special cell adhesion sequence in osteopontin generated in the osteoblast differentiation process, so that bone formation is accelerated.
4. The hydroxyapatite nanowire is inserted into the microsphere after the preparation of the cell microsphere is completed to provide nutrient substances, metabolites and channels for oxygen communication for the internal cells.
5. The results of the examples show that the cell microspheres prepared by the preparation method provided by the invention can be effectively formed, and the cell necrosis of the core region of the cell microspheres is improved.
The technical scheme of the invention is further described in detail through the drawings and the embodiments.
Drawings
FIG. 1 is a photograph of a bright field microscope of the cell microspheres prepared in comparative example 1 and examples 1 and 2 of the present invention; wherein a is a bright field microscope picture of the cell microsphere prepared in comparative example 1, b is a bright field microscope picture of the cell microsphere prepared in example 1, c is a bright field microscope picture of the cell microsphere prepared in example 2;
FIG. 2 is a photograph of apoptotic cell staining of the cell microspheres prepared in comparative example 1 and examples 1 and 2 according to the present invention; wherein a is an apoptotic cell staining picture of the cell microsphere prepared in comparative example 1, b is an apoptotic cell staining picture of the cell microsphere prepared in example 1, c is an apoptotic cell staining picture of the cell microsphere prepared in example 2;
FIG. 3 is a fluorescent image of osteopontin of the cell microspheres prepared in comparative example 1 and examples 1 and 2 of the present invention; wherein a is the osteopontin fluorescence image of the cell microsphere prepared in comparative example 1, b is the osteopontin fluorescence image of the cell microsphere prepared in example 1, and c is the osteopontin fluorescence image of the cell microsphere prepared in example 2.
Detailed Description
Unless defined otherwise, technical or scientific terms used herein should be given the ordinary meaning as understood by one of ordinary skill in the art to which this invention belongs.
Example 1
A method for preparing a cell microsphere, comprising the steps of:
(1) Sterilizing the hydroxyapatite nanowire: soaking the prepared hydroxyapatite nanowire powder in ethanol with the concentration of 99.5%, wherein the ultraviolet irradiation time is 5 hours, cleaning and centrifuging for three times by using PBS after the irradiation is finished, cleaning and centrifuging for one time by using a complete culture medium, wherein the centrifugation revolution is 1500r, and the centrifugation time is 5 minutes, so as to obtain the sterilized hydroxyapatite nanowire powder;
(2) Taking the sterilized hydroxyapatite nanowire powder in the step (1), and adding a complete culture medium to prepare a hydroxyapatite nanowire suspension with the concentration of 0.02 mug/mu L;
(3) Cell suspension preparation: the adherent cells in the culture dish are subjected to trypsin digestion, then are blown and centrifuged, the supernatant is discarded, and the culture medium is added for blowing and beating to obtain the culture dish with the concentration of 4 multiplied by 10 4 Cell suspensions per mL.
(4) Mixing the suspension prepared by the sterilized hydroxyapatite nanowire in the step (2) with the cell suspension prepared in the step (3) in a pipetting mode to obtain a mixed suspension, wherein the volume ratio of the two suspensions is 1:1, a step of;
(5) Inoculating the mixed suspension in the step (4) into a low-adsorption 96-well plate, adding 200 mu l of the mixed suspension into each well, and incubating cells and hydroxyapatite nanowires in the mixed suspension for 48 hours to form cell microspheres, wherein the incubation condition is 37 ℃ and 5% CO 2 The culture medium is replaced once in 2 days, and the replacement volume ratio is 50%.
Example 2
The difference from example 1 is that the concentration of the hydroxyapatite nanowire suspension in step (2) is 0.04. Mu.g/. Mu.L, and the rest is the same as in example 1.
Comparative example 1
The difference from example 1 is that the suspension prepared by sterilizing the hydroxyapatite nanowire in step (4) is replaced with a complete medium, and the rest is the same as example 1.
Characterization of results
1. The cell microspheres prepared in example 1 and the cell microspheres prepared in example 2 and comparative example 1 were subjected to sphere formation observation using a Nikon ECLIPSE Ts2 inverted microscope, and the characterization results are shown in fig. 1, wherein a is a bright field microscope picture of the cell microspheres prepared in comparative example 1, b is a bright field microscope picture of the cell microspheres prepared in example 1, and c is a bright field microscope picture of the cell microspheres prepared in example 2.
As can be seen from FIG. 1, the cell microspheres formed in example 1 and example 2 after 48 hours were larger than those in comparative example 1, and the cell microspheres formed in example 2 were larger than those in example 1. At the same time, example 1, example 2 and comparative example 1 all successfully formed cell microspheres, demonstrating that the addition of hydroxyapatite nanowires during the preparation of cell microspheres can effectively form cell microspheres.
2. The apoptotic cells in the cell microspheres prepared in example 1 and the cell microspheres prepared in example 2 and comparative example 1 were stained and software counted using Tunel apoptotic cell staining, and the test results are shown in fig. 2 and table 1.
TABLE 1 apoptotic cell proportion
| Example 1 | Example 2 | Comparative example 1 | |
| Proportion of apoptotic cells (%) | 2.2 | 1.6 | 20.9 |
As can be seen from fig. 2, the number of apoptotic cells in comparative example 1 was the largest, and the number of apoptotic cells in example 1 and example 2 was smaller, compared with example 1 and example 2.
It can be seen from table 1 that the number of apoptotic cells in example 1 is greater than that in example 2, and is smaller than that in comparative example 1, and the number of apoptotic cells in example 2 is smaller than that in comparative example 1. The hydroxyapatite nanowires are inserted into the cell microspheres, so that more substance exchange channels are provided for the cell areas inside the cell microspheres, and the problem of core necrosis of the cell microspheres can be solved.
3. The cell microspheres prepared in example 1 and the cell microspheres prepared in example 2 and comparative example 1 were stained for osteopontin by immunofluorescence staining, and the test results are shown in fig. 3 and table 2 through software statistics.
TABLE 2 average fluorescence intensity of osteopontin
| Example 1 | Example 2 | Comparative example 1 | |
| Average fluorescence intensity | 84.371 | 70.876 | 61.818 |
As can be seen from fig. 3, the average fluorescence intensity of osteopontin represented in comparative example 1 was the lowest compared with examples 1 and 2, and the average fluorescence intensity of osteopontin represented in example 1 was higher compared with examples 1 and 2.
It can be seen from Table 2 that the average fluorescence intensity representing osteopontin in example 1 is higher than that in example 2 and comparative example 1, while the average fluorescence intensity in example 2 is higher than that in comparative example 1. The hydroxyapatite nanowire can enhance the osteogenic differentiation capacity of the cell microsphere.
The cell microsphere prepared by the invention can be used for preparing organoids and biological 3D printing and constructing large-size high-cell-density bionic tissues.
Therefore, the cell microsphere and the preparation method thereof overcome the problems of complex preparation process and low cell activity of the cell microsphere in the prior art, and the cell microsphere can be effectively obtained by co-culturing the hydroxyapatite nanowire and the cell suspension, the activity of the cell microsphere is ensured, and the differentiation capacity of the cell microsphere is enhanced. The cell microsphere can also be used for preparing organoids or for biological 3D printing to build large-size high-cell-density bionic tissues.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention and not for limiting it, and although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that: the technical scheme of the invention can be modified or replaced by the same, and the modified technical scheme cannot deviate from the spirit and scope of the technical scheme of the invention.
Claims (6)
1. A method for preparing a cell microsphere, comprising the steps of:
(1) Sterilizing the hydroxyapatite nanowire: soaking the prepared hydroxyapatite nanowire powder in ethanol, irradiating with ultraviolet rays, washing and centrifuging for three times by using PBS after the irradiation is finished, and washing and centrifuging once by using a complete culture medium to obtain sterilized hydroxyapatite nanowire powder;
(2) Taking the sterilized hydroxyapatite nanowire powder in the step (1), and adding a complete culture medium to prepare a hydroxyapatite nanowire suspension;
(3) Cell suspension preparation: the adherent cells in the culture dish are subjected to trypsin digestion and then are blown and centrifuged, supernatant is discarded, and a complete culture medium is added for blowing and beating to obtain a cell suspension;
(4) Mixing the suspension prepared by the sterilized hydroxyapatite nanowire in the step (2) with the cell suspension prepared in the step (3) in proportion to obtain a mixed suspension;
(5) Inoculating the mixed suspension in the step (4) into a low-adsorption 96-well plate, adding the mixed suspension into each well, and incubating cells and hydroxyapatite nanowires in the mixed suspension to form cell microspheres;
wherein the hydroxyapatite nanowire in the step (1) has a bundled array structure, the diameter of the hydroxyapatite nanowire is 30-50 nm, and the length of the hydroxyapatite nanowire is 10-25 mu m;
the concentration of the hydroxyapatite nanowire suspension in the step (2) is 0.01-0.08 mug/mu L;
the volume ratio of the cell suspension to the hydroxyapatite nanowire suspension in the step (4) is 1:1-1.5.
2. The method according to claim 1, wherein the concentration of ethanol in the step (1) is 95-99.5%.
3. The preparation method according to claim 1, wherein the ultraviolet irradiation time in the step (1) is 3 to 5 hours; the centrifugal revolution is 1000-1500 r, and the centrifugal time is 3-5 min.
4. The method according to claim 1, wherein the concentration of the cell suspension in the step (3) is 4X 10 4 ~5×10 4 The cell type is one of bone marrow mesenchymal stem cells, osteoblast precursor cells and embryonic stem cells.
5. The method according to claim 1, wherein the amount of the mixed suspension added to each well of the low adsorption 96-well plate in the step (5) is 150 to 200. Mu.L, the incubation temperature is 35 to 37 ℃, and the temperature is CO 2 The concentration is 4.5-5%, the incubation and standing time is 24-48 h, the culture medium is replaced for 2-3 days once, and the volume ratio of the replaced culture medium is 20-50%.
6. The cell microsphere prepared by the preparation method according to any one of claims 1 to 5.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090220561A1 (en) * | 2005-04-28 | 2009-09-03 | Sungho Jin | Compositions comprising nanostructures for cell, tissue and artificial organ growth, and methods for making and using same |
| CN115636400A (en) * | 2022-11-04 | 2023-01-24 | 山东大学 | Preparation method of two-stage structure one-dimensional multifunctional hydroxyapatite nanobelt and application of two-stage structure one-dimensional multifunctional hydroxyapatite nanobelt in assembling functional stem cell ball |
| US20230136323A1 (en) * | 2020-03-19 | 2023-05-04 | Kunimine Industries Co., Ltd. | Spheroid formation promoter |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090220561A1 (en) * | 2005-04-28 | 2009-09-03 | Sungho Jin | Compositions comprising nanostructures for cell, tissue and artificial organ growth, and methods for making and using same |
| US20230136323A1 (en) * | 2020-03-19 | 2023-05-04 | Kunimine Industries Co., Ltd. | Spheroid formation promoter |
| CN115636400A (en) * | 2022-11-04 | 2023-01-24 | 山东大学 | Preparation method of two-stage structure one-dimensional multifunctional hydroxyapatite nanobelt and application of two-stage structure one-dimensional multifunctional hydroxyapatite nanobelt in assembling functional stem cell ball |
Non-Patent Citations (1)
| Title |
|---|
| 梁瑜等: "三维成球优化间充质干细胞的研究进展", 《中国材料进展》, vol. 39, no. 4, pages 281 - 282 * |
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