Genetic variations of bone marrow mesenchymal stromal cells derived from acute leukemia and myelodysplastic syndrome by targeted deep sequencing

Characterization by Next Generation Sequencing of mutations of spliceosome genes and bone marrow mesenchymal cells in patients with myelodysplastic syndromes

“…In our study we tried to reveal whether MSC from MDS patients harbor molecular alterations, and whether these potential abnormalities could be similar to those present in hematopoietic progenitor cells presented. The NGS analysis in 58 MDS patients revealed that gene mutations are more frequent in hematopoietic cells (HPC) than in mesenchymal stromal cells (MSC) confirming previous results in a short number of MDS patients [105], although some papers reported no mutations in MCS maybe because the low number of MDS patients analyzed. [104,150,151].…”

“…Moreover a focused study in 8 genes in 7 MDS patients failed to demonstrate MSC mutations [104]. By contrast, an additional study, carried out in only five MDS patients, showed some mutations in the MSC [105]. Therefore new studies in large series of patients are needed to define the genetic abnormalities of MSC in MDS patients.…”

Downregulation of extracellular vesicle microRNA‑101 derived from bone marrow mesenchymal stromal cells in myelodysplastic syndrome with disease progression

“…Development of MDS is known to be associated with the accumulation of genetic abnormalities by aging, so-called 'Clonal Hematopoiesis of Indeterminate Potential (CHIP)', and additional mutations in critical genes, including epigenetic regulator genes, accelerate AML (34). In addition to genetic abnormalities in MDS cells, BM-MSCs are also shown to have synonymous or non-synonymous mutations (15,35,36) such as 'genetic injury.' The unresolved issue is whether demethylating agents available for patients with MDS could exert any effect on BM-MSCs, especially BM-MSC-derived EV, as miR-101 targets many epigenetic regulator genes.…”

“…Culture of BM-MSCs. BM-MSCs from patients with MDS and AML/MRC were isolated using the conventional plastic adhesion method with a minor modification (15). Briefly, 0.5 to 1 ml of freshy obtained heparinized BM aspirates were cultured in equivalent volumes of Roswell Park Memorial Institute (RPMI)-1640 medium (Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS; HyClone) and 1% penicillin-streptomycin (P/S; Thermo Fisher Scientific, Inc.) and Dulbecco's modified Eagle's medium (DMEM) (Thermo Fisher Scientific) containing 10% FBS (GE Healthcare), 1% of P/S, and 1% non-essential amino acids (NEAAs; Thermo Fisher Scientific, Inc.).…”

“…Cells were cultured for 3 to 5 days, and the medium was changed to DMEM (with 10% of FBS, 1% of P/S and 1% of NEAAs) for non-hematopoietic expansion after the removal of non-adherent cells. Cultured BM-MSC popula-tion was identified as CD34 − /CD45 − /CD73 + /CD90 + /CD105 + with flow cytometry with <5% CD34 + and CD45 + (15).…”