TY - JOUR
T1 - A Rapid Protocol for Direct Isolation of Osteoclast Lineage Cells from Mouse Bone Marrow
AU - Dang, Lei
AU - Li, Nanxi
AU - Wu, Xiaohao
AU - Li, Dijie
AU - Zhang, Zongkang
AU - Zhang, Bao Ting
AU - Lyu, Aiping
AU - Chen, Lin
AU - Zhang, Ge
AU - Liu, Jin
N1 - Funding Information:
This work was supported by the funds from National Key R&D Program of China (2018YFA0800804 to L.C.) and National Natural Science Foundation Council of China (81702189 to J. L), the Theme-based Research Scheme from the Research Grants Council of Hong Kong (T12-201/20-R to A.L.), the General Research Funds from the Research Grants Council of Hong Kong (12100918, 12114416 and 12101117 to G.Z., 12136616 and 12103519 to J. L, 14112915 to B.-T.Z.), the Basic and Applied Basic Research Fund from Department of Science and Technology of Guangdong Province (2019B1515120089 to G.Z.), the 2020 Guangdong Provincial Science and Technology Innovation Strategy Special Fund (Guangdong-Hong Kong-Macau Joint Lab, 2020B1212030006),the Interdisciplinary Research Clusters Matching Scheme of Hong Kong Baptist University (RC-IRCs/17-18/02 to G.Z.; RC-IRCs/17-18/04 and RC-IRMS/15-16/01 to A.L.), the Inter-institutional Collaborative Research Scheme from Hong Kong Baptist University (RC-ICRS/19-20/01 to A.L.). This protocol was modified from previous work (Liu et al., 2015 and 2021).
Publisher Copyright:
© 2022 The Authors
PY - 2022/3/5
Y1 - 2022/3/5
N2 - Osteoclast lineage cells (OLCs), including osteoclast precursors (OCPs) and mature osteoclasts (MOCs), participate in bone remodeling and mediate pathologic bone loss. Thus, it is essential to obtain OLCs for exploring their molecular features in both physiological and pathological conditions in vivo. However, the conventional protocols for obtaining OLCs ex vivo are not only time-consuming, but also unable to capture the cellular status of OLCs in vivo. In addition, the current antibody-based isolation approaches, such as fluorescence-/ magnetic-activated cell sorting, are not able to obtain pure osteoclasts because no unique surface antigen for osteoclasts has been identified. Here, we develop a rapid protocol for directly isolating OLCs from mouse bone marrow through magnetic-activated cell sorting (MACS). This protocol can rapidly enrich OCPs and MOCs, respectively, depending on the expression of the distinctive surface markers at their differentiation stages. It is optimized to isolate OLCs from four mice concurrently, of which sorting procedure could be completed within ~5 h.
AB - Osteoclast lineage cells (OLCs), including osteoclast precursors (OCPs) and mature osteoclasts (MOCs), participate in bone remodeling and mediate pathologic bone loss. Thus, it is essential to obtain OLCs for exploring their molecular features in both physiological and pathological conditions in vivo. However, the conventional protocols for obtaining OLCs ex vivo are not only time-consuming, but also unable to capture the cellular status of OLCs in vivo. In addition, the current antibody-based isolation approaches, such as fluorescence-/ magnetic-activated cell sorting, are not able to obtain pure osteoclasts because no unique surface antigen for osteoclasts has been identified. Here, we develop a rapid protocol for directly isolating OLCs from mouse bone marrow through magnetic-activated cell sorting (MACS). This protocol can rapidly enrich OCPs and MOCs, respectively, depending on the expression of the distinctive surface markers at their differentiation stages. It is optimized to isolate OLCs from four mice concurrently, of which sorting procedure could be completed within ~5 h.
KW - Calcitonin receptor
KW - Magnetic-activated cell sorting
KW - Osteoclast lineage cells
KW - Osteoclast-associated receptor
KW - Osteoporosis
UR - http://www.scopus.com/inward/record.url?scp=85125954586&partnerID=8YFLogxK
U2 - 10.21769/BioProtoc.4338
DO - 10.21769/BioProtoc.4338
M3 - Journal article
AN - SCOPUS:85125954586
SN - 2331-8325
VL - 12
JO - Bio-protocol
JF - Bio-protocol
IS - 5
M1 - e4338
ER -