STUDIES ON DIFFERENTIATION AND ANTI-APOPTOSIS OF ADULT SD RAT MARROW STORMAL CELLS / 解剖学报

ABSTRACT

Objective In previous experiment we proved that rat marrow stromal cells(MSCs) could be induced to differentiate to neuron\|like cells by ? mercaptoethanol and all\|trans\|retinoic acid(ATRA), but the neuron\|like cells showed apoptosis sooner after the differentiation.In this study, we used several sets of combinations of anti\|apoptotic reagents and inductive reagents to induce the differentiation of MSCs,to prevent the differentiated cells from apoptosis and to elongate the lifetime of the neuron\|like cells which might be use in clinic. Methods The MSCs were isolated primarily from bone marrow of adult rat,and purified by passage culture.The 5th passage of MSCs was induced by single or combinative induction,which included RA,Melatonin(10 -9 mol/L), bFGF,bFGF+RA,Melatonin+RA,Melatonin+bFGF.At 1?h,5?h and 24?h of induction, the ratio of neuron\|like cells and the numbers of survived neuron\|like cells among 6 groups were compared.In addition,immunocytochemical stainings with anti\|MAP\|2 and GFAP were also performed on cells of 5?h of induction. Results Neuron\|like cells were all MAP\|2 positive,but GFAP negative.The ratios of neuron\|like cells in MSCs at 1?h after induced by six combinations of reagents were 0 24,0 28,0,0,0 44,0 64;at 5?h were 0 56,0 28,0,0,0 76,0 83 and at 24?h were 0 10,0 27,0,0 14,0 35,0 59 respectively.The ratios of survived neuron\|like cells of differentiated cells at 24?h after induction by RA,Melatonin,Melatonin+RA and Melatonin+bFGF were 0 08,0 37,0 05,0 51 respectively.Conclusions\ 1.bFGF can't induce MSCs into neuron\|like cells independently.Moreover,it suppressed the differentiation effects of RA,while enhancing the effects of MT.2.MT can induce MSCs differentiating into neuron\|like cells and has the anti\|apoptotic effects on the induced neuron\|like cells.3.When MT and bFGF were added together,not only the ratio of neuron\|like cell in MSCs was increased, the lifetime of neuron\|like cells was also elongated.