Morphological and functional characteristics of short-term and long-term bone marrow cultures in chronic myelogenous leukemia

Abstract

Clonogenic capacity of bone marrow progenitors and stromal layers established from bone marrow of 12 patients with CML and 13 healthy controls were evaluated. The initial BFU-E and CFU-GM contents were slightly higher in the CML patients (p > 0.05) in contrast to CFU-GEMM. CFU-GEMM was lower in the patients compared to healthy controls (p < 0.001). In long-term cultures, the number of non-adherent cell population and total clonogenic progenitor cell content decreased gradually in both groups. Weekly evaluation of stromal confluency of adherent cells revealed that establishment of adherent stromal layer was slower in CML patients than in control samples (p < 0.05). At the end of fourth week, the number of samples presenting confluency was 41.7% in the CML group compared with 92.3% in the controls. The initial CD34 positive cell content of the bone marrow samples was similar in both groups. Although CD34 positive cell number in the adherent stromal layer was well preserved in the control group at the end of 4 weeks, this figure decreased significantly in the CML group. The numbers of total adherent cells as well as the total clonogenic progenitor content of adherent layer were also lower in the CML group (3.03% vs 98.2%). When normal CD34+ cells were cultured on IFN-alpha-treated stromal layer followed by the assessment of the long-term culture initiating cells, a reduced capacity to support hemopoietic growth was observed with IFN-alpha-treated normal stroma. This reduction was even higher when CML stroma was treated with IFN-alpha followed by the seeding of the normal CD34+ cells on this stromal layer (26.9% vs 42.8%). These findings show that stromal cells are abnormal in CML patients as well as the progenitor cells, and IFN-alpha treatment causes further defects of the stromal cells.

Curcumin derivative C817 inhibits proliferation of imatinib-resistant chronic myeloid leukemia cells with wild-type or mutant Bcr-Abl in vitro

“…Long term culture-initiating cells (LTC-ICs) assay [20][21][22][23] M2-10B4 cells and SL/SL fibroblasts were established as feeder layers then irradiated at 60 Gy. MNC cells from CML bone marrow were treated with or without C817, or imatinib for 24 h, and then plated in quinplicate on the irradiated feeder layers in MyeloCult ® H5100 medium supplemented with hydrocortisone at a final concentration of 1 μmol/L.…”

“…LTC-IC is recognized as the most stringent assay to detect very primitive human hematopoietic stem cells in vitro [20][21][22][23] . For successful read-out, stem cells must be fully functional, survive for 5 weeks in the presence of a stromal layer.…”

In vitro effect of adenovirus‐mediated human Gamma Interferon gene transfer into human mesenchymal stem cells for chronic myelogenous leukemia

“…Human BMSCs were isolated and cultured according the methods described previously [31]. Heparinized BM samples were obtained by aspiration from the posterior iliac crest of two healthy volunteers and six CML patients into syringe containing an equal volume of Dulbecco's modified Eagle's medium-low glucose (DMEM-LG) (GIBCO) after informed consents were obtained from these BM donors.…”