深低温冻存时限对成骨细胞生物学特性影响的实验研究

探讨深低温冻存不同时限后复苏对成骨细胞生物学特性的影响。取新生SD大鼠颅骨,胶原酶消化法培养成骨细胞。采用活细胞数、成骨细胞增殖功能、碱性磷酸酶(ALP)活性的测定等对深低温冻存1、3、6个月后复苏的成骨细胞和新鲜培养的细胞进行了比较。发现各组成骨细胞在细胞增殖功能、碱性磷酸酶活性方面均无显著性差异(P>0.05)。深低温冻存6个月后复苏组成骨细胞活细胞数与冻存1、3月后复苏组及新鲜培养的细胞组相比,活细胞数降低,有显著性差异(P<0.05),深低温冻存1、3个月后复苏组的活细胞数与新鲜培养的细胞组相比无显著性差异(P>0.05)。实验结果表明:较长时间深低温冻存能减少复苏后成骨细胞的活细胞数。但经深低温冻存后复苏的细胞仍保持成骨细胞原有的生物学特性。

Experimental Study of the Effects of Time-phased Cryopreservation and Resuscitation on Biological Characters of Osteoblasts

This is an in vitro study to explore the effects of cryopreservation and resuscitation on the biological characteristics of osteoblasts at different times. Osteoblasts taken from the crania of newly born SD rats were cultured. Comparative studies were made on the cells' proliferation, the activity of alkaline phosphatase (ALP), and the number of live cells among fresh cultured osteoblasts and cells after the inception of cryopreservation and resuscitation at time-periods of one, three, six months respectively. The results showed that there were no significant differences among four groups in cell proliferation and in activity of ALP(P>0.05). Yet,after cryopreservation and resuscitation, there were significant differences between the six-month group and the other three groups(P<0.05). The results also showed, after cryopreservation and resuscitation, there were no significant differences between the control group and the one-month and three-month groups, respectively(P>0.05). These findings indicated that the live cells might decrease in number after the osteoblasts were cryopreserved for too long a period, but after cryopreservation and resuscitation, the cells still retained the original biological characteristics of osteoblasts.