Proliferative Capacity Of ADSC Derived From Different Parts Of SD Rat And Study Of CM-dil In Vivo Tracing

Objective: To compare the proliferative capacity of ADSC in SD rats differentparts in vitro; explore the feasibility and timeliness of the fluorescent dye CM-Dil vitromarker ADSC.Method: Under sterile conditions cut the adipose tissue from the groin andepididymal of SD rats in turn. Digest these tissues with type Ⅰ collagenase to observethe ADSC and cultured and passaged by differential adhesion. Observe themorphological changes，cell growth and reproduction and calculate the the number ofstem cells extracted per unit volume (1ml) fat by the same way from the groin area(group A) and epididymis region (the group B), then draw their own growth curve.Take up to80%of the third generation(P3) epididymis fat source sex source of stemcells, divided into the experimental group(a、b and c group) and the coutrol group(dgroup). The use of the different concentrations of CM-Dil mark liquid outer markADSC, respectively for group a (20μ g/ml), group b (30μ g/ml), group c (40μg/ml).There are no CM-Dil mark and othr special processing in the blank control group(group d), and in vitro culture and generation by differential adhesion.Observe andcompare two sets of cells form, draw the group and control group cell growth curve,calculating the mark rate of a,b,c three groups of cells after24hours and analysis thesuit the feasibility of the method. During25days, calculated the mark rate of a, b and cgroup of cells at different time points. Analysis statistically between the qol and discussthe timeliness.Results:①observed under inverted phase contrast microscope, two groups (Aand B) fat stem cells were short spindle.With the incubation time, the cells graduallybecome slender, both cell morphology similar to fibroblasts;②The same digest Get method under group A (groin area) the number of cells is (1.03±0.19)×10~3, and thenumber of group B (epididymis District) cells is (2.08±0.15)×10~3.Both statisticallysignificant difference (P <0.05);③A and B groups ADSC of the growth curve showedthe "S" shape.Group B (the epididymis District) ADSC is more proliferative capacitythan in group A (groin area);④The mark rates of the cells after24hours which waslabeled by different concentrations of CM-Dil labeled liquid (a group:20μg/ml, group b:30μg/ml, group c:40μg/ml) were:(98.16±0.31)%,(98.93±0.77)%,(98.09±0.50)%.There is no statistically significant differences between the two groups (P>0.05);⑤Between the experimental group (20μg/ml,30μg/ml,40μg/ml) and the control group,cell growth curve showed "S"-shaped.Marked of20μg/ml and30μg/ml concentration oncell proliferation and vitality had no significant effect.⑥When cells were generated tothe8th generation (25d), labeled rate of c group (40μg/ml) can be attained (74.37±1.68)%.Compared with the rates of a group (20μg/ml),which is (67.59±2.53)%, there isstatistically significant differences (P <0.05).Conclusion:1. Rat epididymal adipose tissue which contained the number of ADSC, cellviability, proliferation ability is superior to ADSC from the inguinal region, could beused as the ideal source of seed cells.2. CM-Dil in vitro markers of ADSC feasible,20μg/ml,30μg/ml concentration ofthe marker of cell proliferation and vitality had no significant effect.3. Different concentrations of CM-Dil labeling rate of ADSC in the short term hadno significant effect，and40μg/ml concentration can be maintained more than3weeks.