Basic Research About Using Umbilical Cord Blood In Supplementary Therapy After High-dose Chemotherapy Or Radiotherapy To Gynecology Oncology

[Objective] To search for a suitable cytokine combination by culturing umbilical cord blood mononuclear cells (MNC) using different cytokines and observing their proliferation.To evaluate the applicability of MNC expanded in vitro by analyzing the engraftment signs and hematopoietic reconstitute in mouse model of myelosuppression.These researches will provide some insight into the characterization of cord blood MNC and some valuable reference of using umbilical cord blood in supplementary therapy for gynecologic malignant tumor.[Methods] After 9d suspension culturing of mononuclear cells from the same unit cord blood in two different cytokine combination (TPO+FL+IL-6, TPO+FL+IL-6+IL-3), the quantitative change of nucleated cells and CD34~+ cells were observed using flow cytometry.Using the techniques of polymerase chain reaction (PCR) and immunohistochemistry, we examined the engrafting of the MNC in the mice that had received tailvein injection: Sublethally irradiated BABL/c mice were randomized into 4 groups, each group of mice was injected with cord blood, fresh MNC, expanded MNC, normal saline respectively. Control group was established. The survival rates and hematopoietic reconstitution of mice in each group were observed and compared. Human hematopoiesis was detected by means of PCR and immunohistochemistry. The expanded MNC group was divided into 2 subgroups according to the difference of cytokine combination, and the survival rates, hematopoietic reconstitution and human hematopoiesis of the 2 subgroups were compared as well.[Results] The proliferative effect of cytokine combination including IL-3 (8.96 ± 0.52 floded) was higher than the other one (6.28± 0.16 floded), the results were above demonstrated significant difference (P<0.01).All the mice have survived until be executed in the end; As for hematopoietic reconstitution, the data collected from transfused mice were significantly higher than those from control group, and the results were above demonstrated significant difference (P<0.05) ,The short-term hematopoietic reconstitution of mice in cord blood transfused group were better than those in fresh or expanded MNC transfused groups; Both fresh and expanded MNC could be implanted into BABL/c mice to produce donor-recipient chimera and to reconstitute hematopoietic system. There was no significant difference in engrafting rate among the groups (P>0.05). And There was no significant difference in all the signs of the 2 subgroups as well.