Identification of human cells in brain xenografts and in neural co-cultures of rat by in situ hybridisation with Alu probe.
Transplantation of human cells into animal models of neurodegenerative disorders is an important scientific application to analyse the survival and developmental capacity of grafted human cells under in vivo conditions. It is critical, therefore, to have a reliable method to distinguish between human and animal cells. In the present study, we describe a combined in situ hybridisation and immunocytochemistry method for the identification of human cells in cultured rat brain cells and xenografts. The specific Alu probe we utilised, which corresponds to the consensus sequence of human Alu repeats was evaluated by southern blot hybridisation of zoo blot and by in situ hybridisation of primary and neoplastic cells from man, rat, mouse, and hamster. This method allows a definite identification of human cells in neural xenografts and, in combination with additional in situ techniques, a further detection of grafted cells.
Just
Lothar
L
Organ and Tissue Culturing, German Research Centre for Biotechnology, Mascheroder Weg 1, D-38124 Braunschweig, Germany. lju@gbf.de
Timmer
Marco
M
Tinius
Jan
J
Stahl
Frank
F
Deiwick
Andrea
A
Nikkhah
Guido
G
Bader
Augustinus
A
D020087:Alu Elements / Q000235:genetics*; D000818:Animals; D015139:Blotting, Southern / Q000379:methods*; D001854:Bone Marrow Cells / Q000145:classification / Q000166:cytology; D001921:Brain / Q000166:cytology / Q000601:surgery; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D003342:Corpus Striatum / Q000166:cytology* / Q000601:surgery; D006224:Cricetinae; D004247:DNA / Q000032:analysis / Q000145:classification / Q000235:genetics*; D015342:DNA Probes; D005260:Female; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D009474:Neurons / Q000166:cytology* / Q000637:transplantation; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D013045:Species Specificity; D014183:Transplantation, Heterologous
D015342:DNA Probes; D004247:DNA