Method for Regulating Osteogenic Differentiation of Periodontal Ligament Stem Cells Based on Extracellular Matrix

A technology of periodontal ligament stem cells and periodontal ligament cells, applied in the field of regulating osteogenic differentiation of periodontal ligament stem cells based on extracellular matrix

Active Publication Date: 2019-12-27
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In order to overcome the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a method for regulating the osteogenic differentiation of periodontal ligament stem cells based on extracellular matrix, which solves the problem that the periodontal ligament stem cells cannot be obtained through repeated expansion of periodontal ligament stem cells in vitro. The technical problem of dry periodontal ligament stem cells has the characteristics of precise regulation of periodontal ligament stem cells in vitro

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  • Method for Regulating Osteogenic Differentiation of Periodontal Ligament Stem Cells Based on Extracellular Matrix
  • Method for Regulating Osteogenic Differentiation of Periodontal Ligament Stem Cells Based on Extracellular Matrix
  • Method for Regulating Osteogenic Differentiation of Periodontal Ligament Stem Cells Based on Extracellular Matrix

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Embodiment 1

[0141] A method for regulating the osteogenic differentiation of periodontal ligament stem cells based on extracellular matrix, comprising the following steps:

[0142] 1) Isolation and identification of bone marrow cell BMCs and periodontal ligament cell PDLCs

[0143] Use the cancellous bone of the jaw to isolate bone marrow cell BMCs; cut the cancellous bone into pieces with sterile ophthalmic forceps, add α-MEM culture medium, 8-12% fetal bovine serum, and then place it at 36.8°C, 4.9% CO 2 Incubator culture under saturated humidity conditions; orthodontic teeth were obtained to isolate periodontal ligament cell PDLCs; the periodontal ligament was digested with 3 mg / mL type Ⅰ collagenase in a 36.8°C incubator for 58 minutes, and centrifuged at 980 rpm for 4.8 minutes, and the tooth The periodontal membrane was resuspended in fresh α-MEM and inoculated on a culture dish; after 5 days, the adherent cells were bone marrow cells BMCs and periodontal ligament cells PDLCs, and a...

Embodiment 2

[0169] A method for regulating the osteogenic differentiation of periodontal ligament stem cells based on extracellular matrix, comprising the following steps:

[0170] 1) Isolation and identification of bone marrow cell BMCs and periodontal ligament cell PDLCs

[0171] Use the cancellous bone of the jaw to isolate bone marrow cell BMCs; cut the cancellous bone into pieces with sterile ophthalmic forceps, add α-MEM culture medium, 8-12% fetal bovine serum, and then place it at 37.2°C, 5.1% CO 2 Incubator culture under saturated humidity conditions; orthodontic teeth were obtained to isolate periodontal ligament cells PDLCs; periodontal ligament was digested with 3mg / mL type Ⅰ collagenase in a 37.2°C incubator for 62min, and then centrifuged at 1020rpm for 5.2min. The periodontal ligament was resuspended in fresh α-MEM and inoculated on a culture dish; after 7 days, the adherent cells were bone marrow cells BMCs and periodontal ligament cells PDLCs, and about 80% of the cell co...

Embodiment 3

[0197] A method for regulating the osteogenic differentiation of periodontal ligament stem cells based on extracellular matrix, comprising the following steps:

[0198] 1) Isolation and identification of bone marrow cell BMCs and periodontal ligament cell PDLCs

[0199] Use the cancellous bone of the jaw to isolate bone marrow cell BMCs; cut the cancellous bone into pieces with sterile ophthalmic forceps, add α-MEM culture medium, 8-12% fetal bovine serum, and then place it at 37°C, 5.0% CO 2 Incubator culture under saturated humidity conditions; orthodontic teeth were obtained to isolate periodontal ligament cells PDLCs; periodontal ligament was digested with 3 mg / mL type Ⅰ collagenase in a 37°C incubator for 60 minutes, centrifuged at 1000 rpm for 5 minutes, and periodontal ligament The membrane was resuspended in fresh α-MEM and inoculated on a culture dish; 6 days later, the adherent cells were bone marrow cells BMCs and periodontal ligament cells PDLCs, and about 80% of t...

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Abstract

A method for regulating osteogenic differentiation of periodontal ligament stem cells based on extracellular matrix comprises the following steps: 1) isolation and identification of bone marrow cellsBMCs and periodontal ligament cells PDLCs; 2) preparation of acellular bone marrow-derived extracellular matrix B-dECM and acellular periodontal ligament derived extracellular matrix P-dECM; 3) for the acellular bone marrow-derived extracellular matrix B-dECM and the acellular periodontal ligament derived extracellular matrix P-dECM obtained in step 2), performing coating tissue culture plate TCPand the acellular periodontal ligament derived extracellular matrix P-dECM by using type IV collagen alpha-2 chain COL4A2; 4) respectively using small interfering RNA and lentivirus transfection to down-regulate and up-regulate the type IV collagen alpha-2 chain COL4A2, the characteristic of accurately regulating COL4A2 in ECM can be obtained.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for regulating the osteogenic differentiation of periodontal ligament stem cells based on extracellular matrix. Background technique [0002] Periodontitis is one of the oldest and most common diseases of mankind. It is a chronic infectious disease of periodontal support tissue caused by microorganisms in dental plaque, which can lead to inflammation of periodontal support tissue, periodontal pocket formation, progressive loss of attachment and alveolar bone resorption, and eventually cause tooth loosening and was removed. According to statistics, the prevalence and severity of periodontitis increase with age, and the prevalence increases significantly after the age of 35, reaching a peak at the age of 50 to 60. At present, the prevalence rate of periodontitis in my country has exceeded 80%, and it is the first cause of tooth loss in adults in my country. At t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/87C12N15/867
CPCC12N5/0662C12N5/0654C12N15/87C12N15/86C07K14/78C12N2510/00C12N2533/90C12N2800/107C12N2740/15043Y02A50/30
Inventor 武俊杰金作林文艺杨鸿旭王阿娴陈晓东
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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