The biocharacterization of intrasplenic transplantation of gene modified mouse fetal liver cells / 中华消化杂志

Objective To develop a genetically modified fetal liver cells (FLC) based transplantation system that can release therapeutic levels of hematopoietic growth factors into the system circulation which can facilitate treatment of patient receiving cytokine therapy following chemotherapy. Method Examine adeno virus mediated gene transfer to isolated murine FLC and evaluate the biocharacterization of intrasplenic transplantation of gene modified murine FLC. Results Substantial transfection rate of 80%～85% were achieved at a ratio of 50 for 2 hr of exposure. Gene modified FLC (FLC GM) labeled with 111 In were injected into the allogenic mice, spleen, the %ID/g of liver was 20%～25% at 24 hr and 50%～55% at 48 hr after transplantation. In addition, serum concentration of GM CSF in mice with intrasplenic transplantation reached its maximum at 48 hr [(356 ?58 ) pg/ml]. Conclusion Intrasplenic transplantation of FLC GM can be predominantly localized in liver and spleen, and engraft rapidly and maintain normal function, which represent a critical step toward successfully accomplishing liver directed gene therapy.

A Study on Intrasplenic Transplantation of Embryonic Stem Cells

PURPOSE: The authors evaluated the morphological and proliferative properties of embryonic stem (ES) cells following intrasplenic transplantation. The results were compared with those obtained following intrasplenic transplantation of cultured hepatocytes. METHODS: ES cells of blastocysts were collected from superovulated Sprague Dawley rats. Hepatocytes were collected from the liver of 7-week old rats by perfusion of collagenase. The ES cells and hepatocytes were cultured for 6 days and transplanted into the rat spleen. The properties of the ES cells and cultured hepatocytes following transplantation were investigated by morphological methods. RESULTS: ES cells in the culture proliferated faster than hepatocytes, and differentiated to various shaped cells. Following transplantation, ES cells were distributed near the periarterial lymphatic sheath. Cultured hepatocytes gathered chiefly around the trabeculae. On PCNA stain of transplanted ES cells, positive cells appeared on day 7 and became distinct on days 10 and 14. Transplanted hepatocytes showed no PCNA positive cells on day 14. On electron microscopic examination, ES cells differentiated to hepatocyte-like structures on day 10, and became functioning hepatocytes on day 14. Transplanted hepatocytes formed bile canaliculi on day 10, although development of organelles was insufficient on day 14. CONCLUSION: ES cells proliferated faster than cultured hepatocytes. Intrasplenic ES cells proliferated at the germinal center and hepatocytes around the trabecula. ES cells differentiated to cells that had the function of hepatocytes.

Acceleration of the Recovery of Chemotherapy-Induced Immune Suppression by the Intrasplenic Transplantation of GM-CSF Gene-Transfected Fetal Liver Cells / 中国肿瘤生物治疗杂志

After murine fetal liver cells (FLC) were transfected with granulocyte-macrophage colony-stimulating factor (GM-CSF) gene by recombinant adenovirus and intrasplenically transplanted into allogeneic mice, the effects of GM-CSF gene-transfected FLC on the recovery of immune response inhibited by chemotherapy were observed. The number of CD4 + cells and the ratio of CD4 + /CDS + cells from peripheral blood lymphocytes increased significantly. The cytotoxicity of the NK cells and the proliferation response of splenocytes to ConA, LPS elevated markedly, but the same results were not from bone marrow. These data demonstrated that intrasplenic transplantation of GM-CSF gene-transfected FLC could effectively accelerate the recovery of immune response after high-dose chemotherapy.