RESUMEN
The purpose of this study was to transplant neonatal rat with human cord blood Lin(-) cells to test the possibility of this xenograft model. The Lin(-) cells were purified from human cord blood (CB) using negative selection strategy based on different lineage-specific antigens. The Lin(-) cells were injected into the liver of neonatal rats using a microinjector at an average of 5 x 10(5) cells for each. Peripheral blood (PB) and spleen were collected at 2,4 and 8 weeks after injection. Flow cytometry was performed to detect human cells in the rat PB, PCR was used to detect human cells in PB as well as spleen. The results showed that a definite proportion of human cells existed in peripheral blood of chimeric rat and the human specific beta2 microglobulin gene fragments were detected in spleen genomic DNA of chimeric rat. It is concluded that human/rat chimera model can be developed with neonatal rats. Human/rat xenograft model may provide a useful and convenient method for human hematopoietic stem cell assay in vivo.
Asunto(s)
Animales , Humanos , Ratas , Animales Recién Nacidos , Trasplante de Células Madre de Sangre del Cordón Umbilical , ADN , Genética , Citometría de Flujo , Antígenos Comunes de Leucocito , Sangre , Reacción en Cadena de la Polimerasa , Ratas Sprague-Dawley , Bazo , Metabolismo , Quimera por Trasplante , Sangre , Genética , Alergia e Inmunología , Trasplante Heterólogo , Microglobulina beta-2 , GenéticaRESUMEN
Bone marrow mesenchymal stem cells (MSCs) are multipotent tissue stem cells that can be induced in vitro to differentiate into a variety of cells such as osteoblasts, chondrocytes and adipocytes. MSCs are useful vehicles for both cell and gene therapy for a variety of diseases. Here, we injected human MSCs with enhanced green fluorescent protein (EGFP) into the striatum of Parkinson disease (PD) rat and examined their survival, migration, differentiation, and the behavior changes in PD rats, which will provide a theoretical foundation and technical method for clinic PD therapy by stem cells. The results showed that human bone marrow MSCs can survive in rat brain for a long time (exceeding 70 d). MSCs were found in multiple areas of the rat brain including the striatum, the corpus callosum, contralateral cortex and even the brain vascular wall. Immunocytochemical staining suggested that implanted cells expressed human neurofilament (NF), neuron-specific enolase (NSE) and glial fibrillary acid protein (GFAP). At the same time, remission in abnormal behavior of the PD rats appeared. Rotation scores decreased gradually from 8.86+/-2.09 r/min pre-transplantation to 4.87+/-2.06 r/min 90 d post-transplantation (statistic result showed P<0.05).