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Culture medium for in-vitro culture of osteoblasts

An in vitro culture, osteoblast technology, applied in the direction of bone/connective tissue cells, culture process, tissue culture, etc.

Inactive Publication Date: 2016-05-11
SUZHOU PULUODA BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, tremella polysaccharides are not used for culturing osteoblasts in vitro

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Low-sugar DMEM 15.0g (HyClone), tremella polysaccharide 100mg / L, calf serum 10mL, appropriate amount of sodium bicarbonate to adjust the pH to 7.2, add water to make up to 1L.

Embodiment 2

[0015] α-MEM 15.0g (HyClone), tremella polysaccharide 200mg / L, fetal bovine serum 8mL, appropriate amount of sodium bicarbonate to adjust the pH to 7.4, add water to make up to 1L.

Embodiment 3

[0017] Enzyme digestion method: take newborn rats, soak them in 75% ethanol, and take the skulls; place the skulls in phosphate buffer solution, remove periosteum, blood vessels and other connective tissues, and cut the washed skulls into small pieces. The bone slices were digested with 0.25% trypsin; after the reaction was terminated, the bone slices were digested with 0.1% type II collagenase to separate the cells; the digested solution containing cells was centrifuged for 10 min, and the precipitated cell aggregates were obtained from Example 1. The culture medium is made into cell suspension, inoculated into cell culture flasks according to the required density, and cultured in a 5% CO2 incubator. After 24 hours, change the medium once, and then change the medium once every two or three days. After culturing for about 12 days, the cells reach confluence and then pass passage.

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PUM

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Abstract

The invention discloses a culture medium for in-vitro culture of osteoblasts. The culture medium is characterized by being prepared from 10.0-15.0 g / L of a basal culture medium, 100-300 mg / L of tremella polysaccharide, 8-10 mL of serum, a proper amount of sodium bicarbonate for regulating the pH value to be 7.2-7.6, and the balance water added for achieving the constant volume of 1 L, wherein the basal culture medium is a DMEM or an alpha-MEM or RPMI-1640 or F12 or DMEM / F12, the purity of the tremella polysaccharide is larger than 75%, and the serum is fetal calf serum or calf serum or human serum or horse serum or sheep serum. The culture medium is used for in-vitro culture of osteoblasts. The culture medium has the advantages that growth of osteoblasts can be accelerated, and the use amount of the serum can be reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a medium for culturing animal cells in vitro, in particular to a medium for culturing osteoblasts in vitro. Background technique [0002] Due to the influence of many factors, the research on bone tissue structure, function and bone metabolism directly through the human body is limited. Therefore, the culture of osteoblasts in vitro, as a commonly used in vitro experimental model, has become an important tool for the study of bone physiology, pathology and repair tissue engineering. Foundation. [0003] In 1964, Peck et al. began to work on the culture of animal embryonic bone osteoblasts. In 1979, Mills et al. first reported that human osteoblasts were successfully cultured in vitro by tissue block culture. Since then, there have been reports on the successful cultivation of osteoblasts. . The main methods are divided into enzymatic digestion method and tissue block method, and the ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0654C12N2500/34C12N2500/60C12N2500/84
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
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