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Construction method of model for detecting cell differentiation functional

A technology for functional models and method establishment, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of unsuitable root slices and high cost

Inactive Publication Date: 2006-12-27
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] (2) Microcapsule encapsulation technology is in the process of research, it may not be suitable for encapsulating the larger root slices in this study, and the required cost is higher
[0007] The existing published technology can observe the effect of EMPs on the attachment and growth of bone marrow stromal cells on the root sheet surface in vitro, but there is no experiment to study its effect on the physiological function of cell differentiation

Method used

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  • Construction method of model for detecting cell differentiation functional
  • Construction method of model for detecting cell differentiation functional
  • Construction method of model for detecting cell differentiation functional

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Materials and methods for isolation and culture of porcine bone marrow stromal cells

[0037] 1. Main experimental materials

[0038] 1. Experimental animals 3-month-old healthy hybrid pigs, weighing about 20kg, provided by the animal room of the Ninth People's Hospital Affiliated to Shanghai Jiaotong University School of Medicine, which meet the national experimental animal quarantine standards.

[0039] 2. Bone marrow puncture supplies Sterile bone marrow puncture bag (with No. 16 bone marrow puncture needle, gauze, hole towel inside), disposable 10ml empty needle, sterile 50ml centrifuge tube, heparin

[0040]3. Liquids and reagents for cell culture

[0041] (1) DMEM culture medium and preparation

[0042] NaHCO 3 3.7g (analytical pure AR, Shanghai Hongguang Chemical Factory, China)

[0043] L-Glutamine 300mg (Sigma, USA)

[0044] Vitamin C 50mg (Sigma, USA)

[0045] Penicillin 200 000u (Shanghai Xinhua Pharmaceutical Factory)

[0046] Streptomycin ...

Embodiment 2

[0086] Materials and methods for extracting enamel matrix protein

[0087] 1. Experimental equipment and reagents

[0088] 1. Main instruments and reagents for enamel matrix protein extraction

[0089] (1) Tooth germ: tooth germ taken from the developing molars in the upper and lower jaws of six-month-old healthy pigs. Healthy hybrid pigs provided by Shanghai Dongfeng Meat Co., Ltd.

[0090] (2) Acetic acid: Wuxi Zhongxi Pharmaceutical Equipment Group Co., Ltd., Jiangsu Province, China, dilute with deionized water when used, and the final concentration is 0.5M / L

[0091] (3) Trichloroacetic acid: analytically pure AR, Jiangsu Yonghua Fine Chemicals Co., Ltd., made into 20% solution with deionized water (w / v, 20g / 100ml)

[0092] (4) Acetone: analytically pure, AR, Suzhou Second Chemical Research Institute Co., Ltd., Jiangsu Province

[0093] (5) Low-temperature high-speed centrifuge: Heraus 28 RS, Germany

[0094] (6) Ultra-low temperature refrigerator (-80°C): REVCO, REVC...

Embodiment 3

[0099] In vivo transplantation experiment of BMSCs compound root slices in nude mice

[0100] 1. Experimental materials

[0101] 1. Subgingival scaler: Gracey spoon curette, Shanghai Kangqiao Dental Medical Equipment Factory

[0102] 2. EDTA gel: 240g / L, Biora AB, Sweden

[0103] 3. Preparation of Bell Lake-Rick Roche decalcification solution

[0104] 70 grams of aluminum chloride crystals

[0105] Distilled water 1000ml

[0106] Formic acid 50ml

[0107] Concentrated hydrochloric acid (37%) 85ml

[0108] After the above ingredients are mixed and dissolved in proportion, set aside.

[0109] 4. Polytetrafluoroethylene microporous film (provided by Shanghai Plastics Research Institute, the brand is H-PSFWM-0.10-90, the thickness is 0.10mm, and the width is 90mm)

[0110] 5. Paraffin embedding machine: Thermo Shandon Histocentre 2, England

[0111] 2. Experimental method

[0112] 1. Obtaining and processing root slices

[0113] The pigs were anesthetized with an overdose ...

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Abstract

The invention relates to a lay down method for model used in detecting cell differentiation function and the method includes the following steps: (1) preparing heelpiece, enamel matrix protein (EMPs) and myeloid stroma cell; (2) covering the heelsurface of the said heelpiece with EMPs; (3) inoculating the myeloid stroma cell on the heelpiece surface obtained by step (2); (4) placing the heelpiece obtained by step (3) into the politef film pocket; (5) placing the heelpiece covered by the pocket under naked mouse skin. The advantages of this invention include connecting the cell culture in vitro and growth in vivo, insulating the impact of naked mouse cell with screen film and inspecting the impact of EMPs to the marrow stroma cell differentiation, which consequently can provide effective research model for studying EMPs application mechanism. At the 8th week of the experiment, different histological structures are formed by different processing groups and the cellular cementum samples are formed on the heelsurface trained by EMPs, which indicates that the EMPs can improve the marrow stroma cell differentiation to the cementoblastoma and also indicates that the research model is successful.

Description

technical field [0001] The invention relates to a method for establishing a model, in particular to a method for establishing a model for detecting cell differentiation functions. Background technique [0002] When cells grown in vitro are implanted back into the body, the cells can go on to form tissues that represent their source. Therefore, it is widely used as a research model to implant the cells cultured in vitro back into the body to observe the function, origin and influencing factors of the cells. Scaffold materials for cell transplantation include synthetic materials and natural materials. In the field of oral cavity research, researchers have used tooth roots or root slices to carry cells. However, the researchers used in vitro co-culture to observe the tissue formed by the cells in vitro, but did not implant the cells bearing the root sheet into the body. Some researchers implanted the tooth root with attached cells into the palatal alveolar bone, which still ...

Claims

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Application Information

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IPC IPC(8): C12N5/06C12Q1/04C12N5/077
Inventor 束蓉宋爱梅谢玉峰宋忠臣王晓春
Owner SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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