TY  - JOUR
TI  - Isolation of Human Bone Marrow Non-hematopoietic Cells for Single-cell RNA Sequencing
AU  - Li, Hongzhe 
AU  - Bräunig, Sandro 
AU  - Scheding, Stefan 
VL  - 14
IS  - 12
PY  - 2024
DA  - 2024/06/20
SP  - e5020
C1  - Bio-protocol 2024;14:e5020
DO  - 10.21769/BioProtoc.5020
UR  - https://doi.org/10.21769/BioProtoc.5020
AB  - The intricate composition, heterogeneity, and hierarchical organization of the human bone marrow hematopoietic microenvironment (HME) present challenges for experimentation, which is primarily due to the scarcity of HME-forming cells, notably bone marrow stromal cells (BMSCs). The limited understanding of non-hematopoietic cell phenotypes complicates the unraveling of the HME’s intricacies and necessitates a precise isolation protocol for systematic studies. The protocol presented herein puts special emphasis on the accuracy and high quality of BMSCs obtained for downstream sequencing analysis. Utilizing CD45 and CD235a as negative markers ensures sufficient enrichment of non-hematopoietic cells within the HME. By adding positive selection based on CD271 expression, this protocol allows for selectively isolating the rare and pivotal bona fide stromal cell population with high precision. The outlined step-by-step protocol provides a robust tool for isolating and characterizing non-hematopoietic cells, including stromal cells, from human bone marrow preparations. This approach thus contributes valuable information to promote research in a field that is marked by a scarcity of studies and helps to conduct important experimentation that will deepen our understanding of the intricate cellular interactions within the bone marrow niche.
KW  - Human bone marrow microenvironment
KW  - Non-hematopoietic cells
KW  - Bone marrow stromal cells
KW  - Flow cytometry
KW  - CD271
KW  - CD45
KW  - CD235a
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False