Method for inducing and disintegrating human amniotic mesenchymal cell to form osteocyte or cartilage cell

Abstract

The invention relates to a method for inducing and disintegrating human amniotic mesenchymal cell to form osteocyte or cartilage cell. The method comprises the following steps: separating human amniotic tissue from placenta; cleaning the human amniotic tissue; digesting the cleaned human amniotic tissue; separating the human amniotic tissue into single human amniotic mesenchymal cell; performing amplification of the human amniotic mesenchymal cell to reach the desired number, adding human amniotic mesenchymal cell suspending solution in high sugar DMEM culture medium with 5% of FBS, 10ng / ml of TGF-beta1, 10ng / ml of IGF, 0.1mu M of decamethasone and 50ng / ml of ascorbic acid to perform induction culture for 21 days while changing the culture medium once per 24h and obtaining the osteocyte or cartilage cell. By using the method of the invention, the osteocyte or cartilage cell can be obtained to repair the bone tissue or cartilage tissue in human body so as to cure the bone defect.

Description

technical field [0001] Since human amnion mesenchymal cells (human Amnion mesenchymal cells, hAMCs) have stem cell biological characteristics such as proliferation activity and multi-differentiation potential, the present invention relates to a method for inducing human amnion mesenchymal cells to differentiate into bone or chondrocytes . Background technique [0002] Avascular necrosis of the femoral head (AVN avascular necrosis of the femoral head), also known as aseptic necrosis of the femoral head (ANFH aseptic necrosis of femoral head) or ONFH osteonecrosis of the femoral head, is a refractory orthopedic disease. Diseases, the use of corticosteroids (corticosteroid, hereinafter referred to as hormones) and excessive drinking in my country are one of the main causes of non-traumatic ONFH. Necrosis of the femoral head is a progressive and disabling disease, most of which end in joint collapse and require artificial joint replacement. Currently commonly used methods inclu...

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Problems solved by technology

Currently commonly used methods include: drugs (inhibiting osteoclastosis, promoting osteogenesis), drilling decompression, autologous or allogeneic fibula transplantation, compression bone grafting, circumflex iliac artery grafting, acetabular rotational osteotomy, stem cell transplantation surgery, etc., but no method can effectively prevent the process of femoral head collapse

In theory, stem cell transplantation is an ideal method to delay the collapse of the femoral head, but due to the limited amount of autologous bone marrow stromal stem cells, the curative effect is uncertain

Although in vitro culture and replantation can obtain a large number of stem cells, they are tumorigenic and difficult to obtain clinical application.

In addition to BMSCs, some scholars have induced embryonic stem cells (ESCs) to differentiate into bone and chondrocytes, but teratomas and immune rejection are likely to occur after allogeneic transplantation. In addition, the application of embryos also involves laws and ethics. problems, which limit their wide application in clinical

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