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In vitro amplification culture method for rabbit subchondral bone osteoblasts, and application thereof

A subchondral bone and culture method technology, applied in the direction of bone/connective tissue cells, animal cells, vertebrate cells, etc., can solve the problem that the isolation effect of subchondral bone osteoblasts is rarely reported, and can meet the requirements of large-scale preparation and acquisition. The effect of short time and high cell viability

Inactive Publication Date: 2012-06-27
TIANJIN HOSPITAL
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Problems solved by technology

Both methods can obtain a certain amount of osteoblasts from the digestion of skull or periosteum-derived tissues of experimental animals. However, there are few reports on the isolation of osteoblasts from subchondral bone in adult arthritis model animals.

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  • In vitro amplification culture method for rabbit subchondral bone osteoblasts, and application thereof
  • In vitro amplification culture method for rabbit subchondral bone osteoblasts, and application thereof
  • In vitro amplification culture method for rabbit subchondral bone osteoblasts, and application thereof

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Embodiment Construction

[0028] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments.

[0029] 1) Improved Hulth animal model [Wang Yunfeng, Bai Renxiao, Zhang Yang, etc. Experimental study of modified Hulth model replicating knee osteoarthritis at different stages. Journal of Tianjin Medical University. 2009, 15(3): 25-27] to construct rabbit joint instability Model: Choose healthy New Zealand big-eared white rabbits (clean grade, Academy of Military Medical Sciences) aged 3-4 months, male or female, intramuscularly inject 100 μl / kg of ketamine (Academy of Military Medical Sciences), Sumianxin II 100 μl / kg (Academy of Military Medical Sciences) ) combined anesthesia, under routine aseptic conditions, a longitudinal incision was made on the anteromedial side of the rabbit knee joint to expose the knee joint, cut off the anterior cruciate ligament, resect the medial meniscus, and cut off the medial collateral ligament. Protect the articul...

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Abstract

The present invention relates to an in vitro amplification culture method for rabbit gonarthritis subchondral bone osteoblasts, and an application thereof. The culture method comprises that: after an improved Hulth rabbit joint instability model is prepared for 4-6 weeks, an aseptic operation is performed to separate the rabbit knee joint; tibial plateau subchondral bone cortex microsclere is digested for 0.5-2 hours by using I type collagenase-EDTA composite enzyme, and is inoculated in a 12-well plate coated by serum; a culture solution is added in batches to carry out culture; after the sclerite attaches for 7-14 days, the cells begin to grow from the edge of the sclerite, wherein the obtained cells have high purity and good proliferation ability. According to the present invention, the method of the present invention is simple and feasible; the obtaining time is short; the obtained cells have high survival rate and high purity; the requirement of the large-scale preparation in a short time can be met; and the important significances are provide for studies of the occurrence and the development of the osteoarthritis, and the prevention and the treatment of the osteoarthritis.

Description

technical field [0001] The invention relates to a method for expanding and culturing subchondral bone osteoblasts in vitro of rabbit knee arthritis and its application. technical background [0002] Osteoarthritis (OA) is a common clinical senile degenerative disease, with joint pain and joint dysfunction (including joint deformity) as the main clinical manifestations. OA is a bone remodeling characterized by cartilage degradation and subchondral sclerosis, and the relationship between the two is of great significance in suggesting changes in the metabolic activity of bone cells. In the treatment of OA, both cartilage changes and subchondral bone degeneration should be prevented. However, most of the studies over the years have focused on cartilage lesions, and there are few studies on subchondral bone. Radin et al first proposed that subchondral bone may play a key role in the initiation and progression of cartilage degeneration. Subchondral bone sclerosis caused by abno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12Q1/02
Inventor 李秀兰张杨郭悦孙晓雷张玮崔丽冯俊奇
Owner TIANJIN HOSPITAL
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