Preparation method of three-dimensional histioid cardiac muscular tissue for studying simulated microgravity effect

Abstract

The invention relates to a preparation method of a three-dimensional histioid cardiac muscular tissue for studying a simulated microgravity effect, in particular relating to a preparation method of a three-dimensional histioid cardiac muscular tissue microballoon, aiming at solving the problems that according to the current cell three-dimensional culture method for studying the simulated microgravity effect, the cytodex microcarrier material is toxic and is not conducive to generation of cardiocytes, a great number of cells are needed for culture, cell harvesting is difficult, the cells are prone to interference of shearing force when being cultured in a klinostat and the continual contraction time of the cardiocytes is short. The method comprises the following steps: preparing a mixed solution; preparing cardiocytes; preparing a cell suspension, adding the cell suspension to a divalent positive ion solution to obtain microballoons; culturing the mircoballons in a culture medium to form the three-dimensional histioid cardiac muscular tissue. The three-dimensional histioid cardiac muscular tissue prepared by the method prepared by the invention can be cultured on a long-term basis,needs no passage and can maintain contraction of the cardiac muscular tissue for a long time. The method is applied to the research fields of space biology, aerospace medicine and tissue engineering.

Description

technical field

[0001] The invention relates to a method for preparing a three-dimensional myocardium-like tissue. Background technique

[0002] The outer space environment has the characteristics of microgravity, ultra-low temperature, high vacuum and strong radiation. Among them, microgravity has a significant impact on human physiological functions. Aerospace medical researchers at home and abroad have conducted a large number of medical observations and studies on the effects of microgravity on human body functions, and found that long-term space flight can cause a certain degree of pathological changes in physiological functions, mainly including cardiovascular system, muscular system, immune system and Pathological changes of the skeletal system. Since the cardiovascular system maintains the normal life activities of the human body, it is of great significance to study the influence of microgravity on the human cardiovascular system, explore its mechanism and formulat...

Examples

specific Embodiment approach 1

[0015] Specific Embodiment 1: In this embodiment, a method for preparing a three-dimensional myocardium-like tissue for simulating microgravity effects is carried out according to the following steps: 1. A sodium alginate solution with a mass concentration of 0.05% to 3% is mixed with a mass concentration of Mixing 10% to 50% type I collagen solution to obtain a mixed solution; 2. Place the heart of a Wistar rat suckling mouse 1 to 3 days after birth in a 0.1mol / L PBS solution at 0 to 4°C, and then Wash the heart with 0.1mol / L PBS solution at 0-4°C for 3-5 times, then cut the heart tissue into 1-3mm 3 Tissue pieces of different sizes were washed with 0.1mol / L PBS solution at 0-4°C for 3-5 times, then digested with digestive juice for 10-30min, then added DMEM medium 10 times the volume of the tissue pieces, centrifuged Discard the supernatant, then add DMEM medium to the precipitate to make cell suspension A, inoculate cell suspension A into a culture bottle, adhere to the wal...

specific Embodiment approach 2

[0018] Embodiment 2: This embodiment is different from Embodiment 1 in that: the mass concentration of the sodium alginate solution in step 1 is 0.5%-2%. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0019] Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that the mass concentration of the sodium alginate solution in step 1 is 1%. Others are the same as in the first or second embodiment.

technical field

[0001] The invention relates to a method for preparing a three-dimensional myocardium-like tissue. Background technique

[0002] The outer space environment has the characteristics of microgravity, ultra-low temperature, high vacuum and strong radiation. Among them, microgravity has a significant impact on human physiological functions. Aerospace medical researchers at home and abroad have conducted a large number of medical observations and studies on the effects of microgravity on human body functions, and found that long-term space flight can cause a certain degree of pathological changes in physiological functions, mainly including cardiovascular system, muscular system, immune system and Pathological changes of the skeletal system. Since the cardiovascular system maintains the normal life activities of the human body, it is of great significance to study the influence of microgravity on the human cardiovascular system, explore its mechanism and formulat...

Examples

specific Embodiment approach 1

[0015] Specific Embodiment 1: In this embodiment, a method for preparing a three-dimensional myocardium-like tissue for simulating microgravity effects is carried out according to the following steps: 1. A sodium alginate solution with a mass concentration of 0.05% to 3% is mixed with a mass concentration of Mixing 10% to 50% type I collagen solution to obtain a mixed solution; 2. Place the heart of a Wistar rat suckling mouse 1 to 3 days after birth in a 0.1mol / L PBS solution at 0 to 4°C, and then Wash the heart with 0.1mol / L PBS solution at 0-4°C for 3-5 times, then cut the heart tissue into 1-3mm 3 Tissue pieces of different sizes were washed with 0.1mol / L PBS solution at 0-4°C for 3-5 times, then digested with digestive juice for 10-30min, then added DMEM medium 10 times the volume of the tissue pieces, centrifuged Discard the supernatant, then add DMEM medium to the precipitate to make cell suspension A, inoculate cell suspension A into a culture bottle, adhere to the wal...

specific Embodiment approach 2

[0018] Embodiment 2: This embodiment is different from Embodiment 1 in that: the mass concentration of the sodium alginate solution in step 1 is 0.5%-2%. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0019] Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that the mass concentration of the sodium alginate solution in step 1 is 1%. Others are the same as in the first or second embodiment.

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