Objective To investigate the way to culture human parathyroid cells and to investigate its secretory function. Methods After digested by collagenase, parathyroid cells were isolated to get the original generation cells, then the cells were cultured and passaged, and morphological changes of original generation cells and passage cells were observed on every day. The parathyroid hormone(PTH) level secreted by the original generation cells and passage cells were measured on the 1st, 5th, 10th, 15th, and 20th day(original generation cells only) respectively. Results The cellular morphology was complete after digestion. On the 2nd day, most of the parathyroid cells had adhered and spreaded, on the 3rd day, all cells had spread. There was no very obvious changes on these cells after cultured for 4-15 days. From 16 to 20 days, some parathyroid cells went senescence. On the 1st day, all of the passage cells, which were fusiform and little bigger than those of the original generation cells, had adhered and spreaded. From 2 to 15 days, there was no very obvious changes. The concentration of PTH in original generation cells begin to decreased significantly on the 10th day (P < 0.01). The concentration of PTH in passage cells were all lower than those of original generation cells at the same corresponding time, but there were no significant difference on the PTH level on 5th day and 1st day, 10th day and 5th day, 15th day and 10th day in passage cells (P > 0.05). Conclusion Parathyroid cells which were cultured within 10 days possess well morphologic structure and have the strongest secretory function. Although the passage cells still possess secretory function, it is greatly inferior to original generation cells. At last, we consider that original generation cells cultured within 10 days can be regarded as the source of allogeneic cell transplantation.
ObjectiveTo explore effect and mechanism of the carcinoma associated fibroblasts (CAFs) of breast cancer on growth and metastasis of breast cancer induced in nude mice by inoculation of CAFs and breast cancer cells. MethodsBreast cancer cell line of MDA-MB-231 (abbreviated as MDA), CAFs, and normal breast tissue fibroblasts (NFs) of the same breast cancer patient were collected, and mixed with normal saline (NS) or SDF-1 ligand blockers of four nitrogen heterocyclic fourteen alka (AMD3100, abbreviated as AMD) for inoculation of nude mice in vivo. According to the different combination, 36 nude mice were randomly divided into 6 groups:MDA+NS group, NFs+NS group, MDA+NFs+NS group, MDA+NFs+AMD group, MDA+CAFs+AMD group, and MDA+CAFs+NS group. Forty six days after the inoculation and feeding, volume of tumor, metastasis of lymph node, lung or liver were observed. In addition, level of plasma SDF-1 was tested by using ELISA method, and expressions of SDF-1 mRNA and protein in tumor specimens were detected by using real-time PCR and Western blot method respectively. ResultsExcept for NFs+ NS group, implanted tumor could be seen in nude mice of other 5 groups. In MDA+CAFs+NS group, the volume of tumor[(9.092±2.662) cm3], level of plasma SDF-1[(75.25±16.23) ng/L], and expression levels of SDF-1 mRNA (the median level was 14.714) and protein (the median level was 0.673). of tumor tissue were significantly greater or higher than those of the other 5 groups (P < 0.050). In addition, lymph node metastasis were found in 4 mice in MDA+CAFs+NS group, and 2 in MDA+NS group. The tumor metastasis of lung and liver was not found in all nude mice. ConclusionsCAFs can promote growth and lymph node metastasis of breast cancer, whose mechanism is related with SDF-1 secreted by CAFs and SDF-1/CXC chemokine receptor 4 (CXCR4), signal pathway.