The skeletal system provides an important basis for supporting the body, coordinating body movement, controlling mineral homeostasis and hematopoietic production. Osteoclasts are specialized cells derived from the fusion of monocyte/macrophage hematopoietic lineage progenitors, and are essential for bone homeostasis and health maintenance. A large number of experiments have demonstrated that its cell formation and dysfunction are closely related to osteoporosis, fracture healing, osteoarthritis and primary/metastatic bone tumors. Osteoclasts, as "bone eaters", have been widely studied for their roles in bone homeostasis and disease.
Osteoclasts are terminally differentiated cells and cannot be passaged. Under microscope, osteoclasts are characterized by large cell bodies, multinucleated, pseudopodia and protrusions.
In experimental studies, osteoclasts are often obtained by inducing stem cells or monocytes in vitro to differentiate with the help of inducers. This method produces osteoclastoid cells. Osteoclastoid cells are primary or induced cells with osteoclastic properties, which have been widely used in the study of osteoclasts in recent years. The cells obtained by induction method have high purity and large quantity, which can satisfy various biochemical and meristem experiments. There are two common methods for osteoclast induction.
Primary cell induction
Currently, the most common cell line that can be induced to differentiate into osteoclasts is mouse RAW 264.7 cell line. Here, we will take this cell as an example to introduce its induction method.
AW 264.7 can be induced by LPS, RANKL, or M-CSF + RANKL.
1. LPS induction.
The concentration could be 10ng/mL, 30ng/mL, 50ng/mL, 100ng/mL. But 30ng/mL, 50ng/mL, 100ng/mL LPS induction will produce more multinucleated cells, and multinucleated cells through 30ng/mL LPS induction increase significantly in size.