当前位置： SCI文献检索 > CELL TRANSPLANTATION期刊下所有文献 > In vitro labeling strategies for identifying primary neural tissue and a neuronal cell line after transplantation in the CNS.

In vitro labeling strategies for identifying primary neural tissue and a neuronal cell line after transplantation in the CNS.

Abstract：

:Potential labels for identifying embryonic raphe neurons and a clonal, neuronally differentiating, raphe-derived cell line, RN33B, in CNS transplantation studies were tested by first characterizing the labels in vitro. The labels that were tested included 4',6-diamidino-2-phenylindole hydrochloride, 1,1'-dioctadecyl-3,3,3'-tetramethylindocarbocyanine perchlorate, the Escherichia coli lacZ gene, Fast Blue, Fluoro-Gold, fluorescein-conjugated latex microspheres, fluorescein isothiocyanate-conjugated or nonconjugated Phaseolus vulgaris leucoagglutinin, methyl o-(6-amino-3'-imino-3H-xanthen-9-yl) benzoate monohydrochloride, or tetanus toxin C fragment. Subsequently, the optimal in vitro labels for embryonic raphe neurons and for RN33B cells were characterized in vivo after CNS transplantation. In vitro, 1,1'-dioctadecyl-3,3,3'-tetramethylindocarbocyanine perchlorate (DiI) optimally labeled embryonic neurons. The Escherichia coli lacZ gene optimally labeled RN33B cells. Most labels were rapidly diluted in cultures of embryonic astrocytes and proliferating RN33B cells. Some labels were toxic and were often retained in cellular debris. In vivo, DiI was visualized in transplanted, DiI-labeled raphe neurons, but not in astrocytes up to 1 mo posttransplant. DiI-labeled host cells were seen after transplantation of lysed, DiI-labeled cells. beta-Galactosidase was visualized in transplanted, Escherichia coli lacZ gene-labeled RN33B cells after 15 days in vivo. No beta-galactosidase was seen in host cells after transplantation of lysed, lacZ-labeled RN33B cells. The results demonstrate that labels for use in CNS transplantation studies should be optimized for the specific population of donor cells under study, with the initial step being characterization in vitro followed by in vivo analysis. Appropriate controls for false-positive labeling of host cells should always be assessed.

journal_name

Cell Transplant

journal_title

Cell transplantation

authors

Onifer SM,White LA,Whittemore SR,Holets VR

doi

10.1177/096368979300200207

subject

Has Abstract

pub_date

1993-03-01 00:00:00

pages

131-49

issue

eissn

0963-6897

issn

1555-3892

journal_volume

pub_type

杂志文章

在线工具

In vitro labeling strategies for identifying primary neural tissue and a neuronal cell line after transplantation in the CNS.