Treatment of murine hypoparathyroidism with transgenetic hemopoietic stem cell transplantation
- VernacularTitle:转基因造血干细胞移植治疗小鼠甲状旁腺功能低下症的研究
- Author:
Yi ZHOU
;
Lemin LIN
;
Chun SONG
- Publication Type:Journal Article
- Keywords:
Hematopoietic stem cell transplantation;
Gene expression regulation;
Parathyroid hormones;
Mice
- From:
Chinese Journal of Organ Transplantation
1996;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the gene therapy of hypoparathyroidism (HPT) with hemopoietic stem cell s(HSCs) integrated with parathyroid hormone (PTH) gene.Methods Human PTH gene was amplified by polymerase chain reaction from pcDNA 3.1 -PTH templates and inserted into murine stem cell virus (MSCV) vectors. The plasmids recombinated with PTH gene were transfected into PA317 packaging cells and screened for positive clones with Geneticin. The condensed recombinant retroviruses were obtained and used to infect HSCs coming from umbilical cord blood. The infected HSCs were injected into the mice suffering from HPT. The change of symptoms and serum level of PTH and calcium in each group of mice was observed.Results Human PTH gene was proved to be inserted into MSCV vectors and the titres were up to 2?10 7 colony forming unit (CFU)/ml in condensed retroviral solution. The secretion of PTH reached 15 ng per 10 6 cells per 48 h. The wild type viruses were not detected by PCR amplification. The mice suffering from HPT recovered quickly and the serum level of calcium and PTH remained normal for about 3 months after the HSCs recombinated with PTH gene were injected into them. In the groups of mice subject to the injection of the recombinant retroviruses containing PTH gene, the serum level of PTH and Calcium was significantly increased in a short time, and then declined slowly. And the mice took on the manifestation of hypoparathyroidism again. The mice only subject to the implantation of HSCs were all dead within 20 days.Conclusion A long term therapeutic effect could be obtained by infusing the HSCs which were integrated with PTH gene into the mice suffered from hypoparathyroidism.
