Experimental Study On The Correlation Between Bone Marrow Stromal Cell And Acute Leukemia

Objective1. Establishing a method of bone marrow stromal cell (BMSC) isolation and culture in vitro.2. Observe the biological properties of BMSC, compare BMSC of control with acute leukemia (AL) in quantity, morphology and growth rate, and discuss the effect of these difference on acute leukemia.3. The cell growth was measured by cell-count method, comparing BMSC of control with acute leukemia in growth trend, and analyzing the effect of different growth trend on acute leukemia.4. Detecting expression of CD54 (intercellular adhesion molecule-1, ICAM-1) and CD106 (vascular cell adhesion molecule-1, VCAM-1) on bone marrow stromal cell (BMSC) and discussing its relationship with acute leukemia.5. Observe the influence of BMSC on leukemia tumor cell (K562) and discuss its clinical significance.Method1. Mononuclear cells were obtained from 5ml adult human bone marrow by density gradient centrifugation with Ficoll solution(1.077g/ml).BMSC were cultured in Dulbecco's Modified Eagle's Medium with low glucose(LG-DMEM) contain 10% fetus calf serum at number of 2×10⁶/ml, then incubated in 37 ℃, 25%CO₂,100% humidity.2. The morphology of BMSC was observed under phase-contrast microscope andphotos were taken in proper time.3. BMSC was plated in 96-well plates. The cell growth was measured by cell-countmethod.4. The flow cytometer was performed to examine the expression of adhesion molecule CD54 and CD106 on BMSC.5. BMSC of control and acute leukemia were plated in culture flask, After 48h when BMSC stick on the culture flask, K562 was added to it and co-cultured for 48h. The apoptosis rate of K562 was examined by flow cytometer.Result1. BMSC harvested in this study were homogenous population and exhibited a spindle-shaped fibroblastic morphology.2. From the cell growth curve, we can find BMSC of AL was less than control group in quantity, and its ability of proliferation was not as good as control.3. Expression of CD54 and CD106 in new AL and CR was lower than that in control group (P<0.05), but there was no significant difference between AL and CR (P>0.05).4. Co-cultured with control group BMSC, apoptosis rate of K562 was higher than those of AL and blank group (P<0.05).Conclusion1. Compared with control, BMSC of AL has some deficiency in quality and quantity. So there was some deficiency in its ability of hematopoiesis sustaining.2. Abnormal expression of adhesion molecule on AL BMSC resulted in abnormity of haematopoietic stem cell (HSC) in differentiation and growth.3. Normal BMSC can restrain tumor cell (K562) from growth and accelerate it to apoptosis, so we conferred that normal BMSC had some tumor-inhibition function.