title: Detection of cross-sex chimerism in the common marmoset monkey (Callithrix jacchus) in interphase cells using fluorescence in situ hybridisation probes specific for the marmoset X and Y chromosomes.
abstract: Chimerism associated with placental sharing in marmosets has been traditionally analysed using conventional chromosome staining on metaphase spreads or polymerase chain reaction. However, the former technique requires the presence of proliferating cells, whereas the latter may be associated with possible blood cell contamination. Therefore, we aimed to develop a single-cell analysis technique for sexing marmoset cells. We applied fluorescent in situ hybridisation (FISH) to cell nuclei using differentially labelled X and Y chromosome-specific probes. Herein we present the validation of this method in metaphase cells from a marmoset lymphoblastoid cell line, as well as application of the method for evaluation of cross-sex chimerism in interphase blood lymphocytes and haematopoietic bone marrow cells from marmosets of same- and mixed-sex litters. The results show conclusively that haematopoietic cells of bone marrow and leucocytes from blood are cross-sex chimeric when the litter is mixed sex. In addition, single samples of liver and spleen cell suspensions from one individual were tested. Cross-sex chimerism was observed in the spleen but not in liver cells. We conclude that FISH is the method of choice to identify cross-sex chimerism, especially when combined with morphological identification of nuclei of different cell types, which will allow a targeted tissue-specific analysis.
authors.affiliation: Department of Reproductive Biology, German Primate Centre, Goettingen, 37077, Germany.|Departments of Gastroenterology and Endoscopy, Novel Hôpital Civil (NHC), University Hospital Strasbourg, 67000, France.|Corresponding author. Email: edris.wedi@chru-strasbourg.fr.
authors.forename: E
authors.initials: E
authors.lastname: Wedi
authors.affiliation: Institute of Human Genetics, University Hospital, Ludwig-Maximilians-University Munich, 80336 Munich, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Müller
authors.affiliation: Institute of Human Genetics, University Hospital, Ludwig-Maximilians-University Munich, 80336 Munich, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Neusser
authors.affiliation: Reproduction Genetics Unit, Department of Gynaecological Endocrinology and Reproductive Medicine, University of Heidelberg, 69047, Germany.
authors.forename: P H
authors.initials: PH
authors.lastname: Vogt
authors.affiliation: Department of Reproductive Biology, German Primate Centre, Goettingen, 37077, Germany.|Division of Reproductive & Developmental Sciences, Oregon National Primate Research Center, Oregon Health & Science University, 505 NW 185th Avenue, Beaverton, OR 97006, USA.
authors.forename: O Y
authors.initials: OY
authors.lastname: Tkachenko
authors.affiliation: Reproduction Genetics Unit, Department of Gynaecological Endocrinology and Reproductive Medicine, University of Heidelberg, 69047, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Zimmer
authors.affiliation: Institute for Anthropology and Human Genetics, Department Biology II, Biocenter, Ludwig-Maximilians-University Munich, 82152 Planegg-Martinsried, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Smeets
authors.affiliation: Department of Obstetrics and Gynecology, University of Goettingen, Goettingen, 37075, Germany.
authors.forename: H W
authors.initials: HW
authors.lastname: Michelmann
authors.affiliation: Department of Reproductive Biology, German Primate Centre, Goettingen, 37077, Germany.
authors.forename: P L
authors.initials: PL
authors.lastname: Nayudu
keywords: bone marrow cells; lymphocytes