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• W2037355105 abstract "Tissue chimerism was recently described in transplanted organs from female donors into male recipients, by demonstration of the Y-chromosome in tissue-derived cells. It was claimed that these Y-chromosome positive cells were recipient derived. To find out whether the chimeric cells, derived from pregnancies of sons or blood transfusions, could have been present in the solid organs before transplantation, we performed the following study. In situ hybridization for the Y-chromosome was performed on the normal organs (51 kidneys, 51 livers, 69 hearts) from 75 women of the normal population, whose child and blood transfusion status were known. Chimeric cells were found in 13 kidneys, 10 livers and 4 hearts, of 23 women. There was no relation between the child status or the blood transfusion history with the presence of Y-chromosome positive cells. We have for the first time demonstrated that male cells are present in normal kidneys, livers and hearts. Theoretically, these organs could have been used for the transplantation. Therefore, our findings demonstrate that the chimeric cells thus far described in transplantation studies, are not necessarily donor derived, and could have been present in the organs before the transplantation. Tissue chimerism was recently described in transplanted organs from female donors into male recipients, by demonstration of the Y-chromosome in tissue-derived cells. It was claimed that these Y-chromosome positive cells were recipient derived. To find out whether the chimeric cells, derived from pregnancies of sons or blood transfusions, could have been present in the solid organs before transplantation, we performed the following study. In situ hybridization for the Y-chromosome was performed on the normal organs (51 kidneys, 51 livers, 69 hearts) from 75 women of the normal population, whose child and blood transfusion status were known. Chimeric cells were found in 13 kidneys, 10 livers and 4 hearts, of 23 women. There was no relation between the child status or the blood transfusion history with the presence of Y-chromosome positive cells. We have for the first time demonstrated that male cells are present in normal kidneys, livers and hearts. Theoretically, these organs could have been used for the transplantation. Therefore, our findings demonstrate that the chimeric cells thus far described in transplantation studies, are not necessarily donor derived, and could have been present in the organs before the transplantation. Chimerism can be defined as a phenomenon in which cells from one individual are present in another individual. These cells can either be circulating or they can be integrated into the parenchyma. After the first description of its occurrence in transplanted organs in 1965 (1Medawar PB Transplantation of tissues and organs: introduction..Br Med Bull. 1965; 21: 97-99Crossref Scopus (80) Google Scholar), chimerism has grown into a frequently discussed concept, especially in relation to transplantation. Important issues are how chimerism is induced, for instance by damage caused due to rejection, and whether it may enhance recipient tolerance. In 2001, Lagaaijet al. (2Lagaai EL Cramer-Knijnenburg GF van Kemenade FJ van Es LA Bruijn JA van Krieken JH Endothelial cell chimerism after renal transplantation and vascular rejection..Lancet. 2001; 357: 33-37Abstract Full Text Full Text PDF PubMed Scopus (267) Google Scholar) demonstrated the presence of recipient-derived endothelial cells in kidney grafts. In other studies, chimeric tubular epithelial cells were described in transplanted kidneys (3Gupta S Verfaillie C Chmielewski D Kim Y Rosenberg ME A role for extrarenal cells in the regeneration following acute renal failure..Kidney Int. 2002; 62: 1285-1290Abstract Full Text Full Text PDF PubMed Google Scholar,4Mengel M Jonigk D Marwedel M et al.Tubular chimerism occurs regularly in renal allografts and is not correlated to outcome..J Am Soc Nephrol. 2004; 15: 978-986Crossref PubMed Scopus (39) Google Scholar). Chimerism was also reported in other transplanted organs: Chimeric endothelium, duct epithelium and hepatocytes were found in transplanted livers (5Fogt F Beyser KH Poremba C Zimmerman RL Khettry U Ruschoff J Recipient-derived hepatocytes in liver transplants: a rare event in sex-mismatched transplants..Hepatology. 2002; 36: 173-176Crossref PubMed Scopus (70) Google Scholar, 6Gao Z McAlister VC Williams GM Repopulation of liver endothelium by bone-marrow-derived cells..Lancet. 2001; 357: 932-933Abstract Full Text Full Text PDF PubMed Scopus (148) Google Scholar, 7Kleeberger W Rothamel T Glockner S Flemming P Lehmann U Kreipe H High frequency of epithelial chimerism in liver transplants demonstrated by microdissection and STR-analysis..Hepatology. 2002; 35: 110-116Crossref PubMed Scopus (110) Google Scholar); chimeric cardiomyocytes and smooth muscle cells were found in transplanted hearts (8Quaini F Urbanek K Beltram AP et al.Chimerism of the transplanted heart..N Engl J Med. 2002; 346: 5-15Crossref PubMed Scopus (1127) Google Scholar,9Thiele J Varus E Wickenhauser C et al.Mixed chimerism of cardiomyocytes and vessels after allogeneic bone marrow and stem-cell transplantation in comparison with cardiac allografts..Transplantation. 2004; 77: 1902-1905Crossref PubMed Scopus (57) Google Scholar) and chimeric bronchial epithelium and type II pneumocytes were found in transplanted lungs (10Kleeberger W Versmold A Rothamel T et al.Increased chimerism of bronchial and alveolar epithelium in human lung allografts undergoing chronic injury..Am J Pathol. 2003; 162: 1487-1494Abstract Full Text Full Text PDF PubMed Scopus (165) Google Scholar). The differences in the reported amounts of chimeric cells in solid organs after transplantation in various studies are remarkable, ranging from no chimerism (11Glaser R Lu MM Narula N Epstein JA Smooth muscle cells, but not myocytes, of host origin in transplanted human hearts..Circulation. 2002; 106: 17-19Crossref PubMed Scopus (191) Google Scholar) to low (12Laflamme MA Myerson D Saffitz JE Murry CE Evidence for cardiomyocyte repopulation by extracardiac progenitors in transplanted human hearts..Circ Res. 2002; 90: 634-640Crossref PubMed Scopus (391) Google Scholar,13Muller P Pfeiffer P Koglin J et al.Cardiomyocytes of noncardiac origin in myocardial biopsies of human transplanted hearts..Circulation. 2002; 106: 31-35Crossref PubMed Scopus (201) Google Scholar) or even high (1Medawar PB Transplantation of tissues and organs: introduction..Br Med Bull. 1965; 21: 97-99Crossref Scopus (80) Google Scholar) levels of chimeric cells. After the publication of Quaini et al. (8Quaini F Urbanek K Beltram AP et al.Chimerism of the transplanted heart..N Engl J Med. 2002; 346: 5-15Crossref PubMed Scopus (1127) Google Scholar) reporting on chimeric cells in heart transplants, the identity and source of the chimeric cells were heavily debated. It was questioned whether the transplanted hearts could not already have been chimeric before transplantation, due to, for instance, circulating fetal cells or cells derived from previous blood transfusions. It was criticized by Bianchi et al. (14Bianchi DW Johnson KL Salem D Chimerism of the transplanted heart..N Engl J Med. 2002; 346: 1410-1412Crossref PubMed Scopus (20) Google Scholar) that fetal stem cells may persist in the pe-ripheral blood in healthy women as long as 27 years after delivery (15Bianchi DW Zickwolf GK Weil GJ Sylvester S DeMaria MA Male fetal progenitor cells persist in maternal blood for as long as 27 years postpartum..Proc Natl Acad Sci. 1996; 93: 705-708Crossref PubMed Scopus (1061) Google Scholar). In approximately 30% of women who had been pregnant of a son, male cells of fetal origin were found in the circulation up to 38 years after delivery (16Evans PC Lambert N Maloney S Furst DE Moore JM Nelson JL Long-term fetal microchimerism in peripheral blood mononuclear cell subsets in healthy women and women with scleroderma..Blood. 1999; 93: 2033-2037Crossref PubMed Google Scholar). It was argued, therefore, that the detected male cells in the transplanted hearts might be derived from the sons of the female donors, and not from the male recipi-ents. Blood transfusions may be another possible source of chimerism. A study that is often cited is from Lee et al. (17Lee TH Paglieroni T Ohto H Holland PV Busch MP Survival of donor leukocyte subpopulations in immunocompetent transfusion recipients: frequent long-term microchimerism in severe trauma patients..Blood. 1999; 93: 3127-3139Crossref PubMed Google Scholar), who found male circulating cells in seven of ten women who had received blood transfusions because of trauma. Therefore, blood transfusions may be a source of chimeric cells already present in the solid organ graft before transplantation. In most transplantation studies, normal organ controls are lacking or relatively scarce. In 16 studies (2Lagaai EL Cramer-Knijnenburg GF van Kemenade FJ van Es LA Bruijn JA van Krieken JH Endothelial cell chimerism after renal transplantation and vascular rejection..Lancet. 2001; 357: 33-37Abstract Full Text Full Text PDF PubMed Scopus (267) Google Scholar, 3Gupta S Verfaillie C Chmielewski D Kim Y Rosenberg ME A role for extrarenal cells in the regeneration following acute renal failure..Kidney Int. 2002; 62: 1285-1290Abstract Full Text Full Text PDF PubMed Google Scholar, 4Mengel M Jonigk D Marwedel M et al.Tubular chimerism occurs regularly in renal allografts and is not correlated to outcome..J Am Soc Nephrol. 2004; 15: 978-986Crossref PubMed Scopus (39) Google Scholar, 5Fogt F Beyser KH Poremba C Zimmerman RL Khettry U Ruschoff J Recipient-derived hepatocytes in liver transplants: a rare event in sex-mismatched transplants..Hepatology. 2002; 36: 173-176Crossref PubMed Scopus (70) Google Scholar,7Kleeberger W Rothamel T Glockner S Flemming P Lehmann U Kreipe H High frequency of epithelial chimerism in liver transplants demonstrated by microdissection and STR-analysis..Hepatology. 2002; 35: 110-116Crossref PubMed Scopus (110) Google Scholar, 8Quaini F Urbanek K Beltram AP et al.Chimerism of the transplanted heart..N Engl J Med. 2002; 346: 5-15Crossref PubMed Scopus (1127) Google Scholar, 9Thiele J Varus E Wickenhauser C et al.Mixed chimerism of cardiomyocytes and vessels after allogeneic bone marrow and stem-cell transplantation in comparison with cardiac allografts..Transplantation. 2004; 77: 1902-1905Crossref PubMed Scopus (57) Google Scholar, 10Kleeberger W Versmold A Rothamel T et al.Increased chimerism of bronchial and alveolar epithelium in human lung allografts undergoing chronic injury..Am J Pathol. 2003; 162: 1487-1494Abstract Full Text Full Text PDF PubMed Scopus (165) Google Scholar, 11Glaser R Lu MM Narula N Epstein JA Smooth muscle cells, but not myocytes, of host origin in transplanted human hearts..Circulation. 2002; 106: 17-19Crossref PubMed Scopus (191) Google Scholar, 12Laflamme MA Myerson D Saffitz JE Murry CE Evidence for cardiomyocyte repopulation by extracardiac progenitors in transplanted human hearts..Circ Res. 2002; 90: 634-640Crossref PubMed Scopus (391) Google Scholar, 13Muller P Pfeiffer P Koglin J et al.Cardiomyocytes of noncardiac origin in myocardial biopsies of human transplanted hearts..Circulation. 2002; 106: 31-35Crossref PubMed Scopus (201) Google Scholar,18Grimm PC Nickerson P Jeffery J et al.Neointimal and tubu- lointerstitial infiltration by recipient mesenchymal cells in chronic renal-allograft rejection..N Engl J Med. 2001; 345: 93-97Crossref PubMed Scopus (244) Google Scholar, 19Hove WR van Hoek B Bajema IM Ringers J van Krieken JH Lagaaij EL Extensive chimerism in liver transplants: vascular endothelium, bile duct epithelium, and hepatocytes..Liver Transpl. 2003; 9: 552-556Crossref PubMed Scopus (69) Google Scholar, 20Sinclair RA Origin of endothelium in human renal allografts..Br Med J. 1972; 4: 15-16Crossref PubMed Scopus (58) Google Scholar, 21Starzl TE Demetris AJ Trucco M et al.Chimerism and donor- specific nonreactivity 27 to 29 years after kidney allotransplantation..Transplantation. 1993; 55: 1272-1277Crossref PubMed Scopus (295) Google Scholar, 22Williams GM ter Haar A Parks LC Krajewski CA Endothelial changes associated with hyperacute, acute, and chronic renal allograft rejection in man..Transplant Proc. 1973; 5: 819-822PubMed Google Scholar) published on chimerism in solid organ transplantation, the negative controls amount to a total of only 18 organs (4 kidneys and 14 hearts), of which the results are clearly described. In three studies on liver transplantation,negative controls are mentioned to be negative, but the number of control tissues is unclear (5Fogt F Beyser KH Poremba C Zimmerman RL Khettry U Ruschoff J Recipient-derived hepatocytes in liver transplants: a rare event in sex-mismatched transplants..Hepatology. 2002; 36: 173-176Crossref PubMed Scopus (70) Google Scholar,7Kleeberger W Rothamel T Glockner S Flemming P Lehmann U Kreipe H High frequency of epithelial chimerism in liver transplants demonstrated by microdissection and STR-analysis..Hepatology. 2002; 35: 110-116Crossref PubMed Scopus (110) Google Scholar,19Hove WR van Hoek B Bajema IM Ringers J van Krieken JH Lagaaij EL Extensive chimerism in liver transplants: vascular endothelium, bile duct epithelium, and hepatocytes..Liver Transpl. 2003; 9: 552-556Crossref PubMed Scopus (69) Google Scholar). Child and blood transfusion status are never mentioned of the controls. Also of the females who donated the organs, information on child and blood transfusion status is mostly lacking. The previously mentioned studies regard chimerism as a post-transplantation phenomenon. However, chimeric baseline levels, meaning the amount of background chimerism already present before transplantation, were never thoroughly investigated. It is necessary to investigate these chimeric baseline levels to distinguish between 'background chimerism' and t'ransplantation induced chimeris'before drawing conclusions with respect to the immunological role of chimeric cells in transplanted organs. The goal of the present study was to find out whether organs that are frequently used for transplantation, namely, kidneys, livers and hearts, may already contain chimeric cells before transplantation. Therefore, we have searched for the presence of Y-chromosome positive cells in normal kidneys, livers and hearts of 75 women, whose child and blood transfusion status were known. Our results provide essential baseline data for future research on the subject of chimerism in solid organ transplantation. Tissue specimens came from autopsies on women, performed at the Lei-den University Medical Center (LUMC) between 1999 and 2001. Clinical exclusion criteria comprised a history of autoimmune disease, solid organ transplantation, bone marrow transplantation or stem cell transplantation. Permission of the medical-ethical committee of the LUMC was obtained to enquire general practitioners about the child status of women on whom autopsy was performed. We enquired 117 general practitioners about the child status of 154 women, and 81% replied. A definite answer about the child status was obtained for 95 women. Of these, 53 had given birth to at least 1 son, 11 had only daughters and 31 had no children. Tissue specimens of women with sons and of women without children were entered into the study. Data on blood transfusions were obtained from the Department of Immuno- haematology and Blood Bank (the IHB) of the LUMC. Data were available from 1987 onward, and included the number, time and type of transfusion. It appeared that 45 of 75 women who eventually entered the study, had received a blood transfusion. It was known how many nucleated cells are maximally present in any transfusion type, making it possible to estimate the maximum amount of nucleated cells transferred by each transfusion. As some transfusions are a combination of several donors, and as there are an equal number of female and male donors on average, an estimate of the maximum number of male-nucleated cells transferred by each transfusion could be made. To obtain information on blood transfusions in the period before 1987, we contacted the general physicians of all women included in the study. Eighty-four percent replied. However, they provided no additional data to those that were already known from the IHB. Tissue specimens of the heart, liver and kidney were reviewed in hema-toxylin and eosin (H&E) staining for histomorphological lesions and signs of autolysis. Tissue specimens of 9 women were consequently excluded, 7 of whom had sons, and 2 of whom had no children. Because the liver block most of the time also contained a tissue specimen from the spleen, we decided to incorporate the spleen into our study as well. All remaining specimens covered at least an area of 1 × 1 cm. To verify the quality of the tissue samples for the detection of sex chromosomes, in situ hybridization for the X-chromosome was performed on a random selection of various organ samples of 46 non-autolytic specimens: all specimens were positive. Tissue specimens of 75 women were entered into the study; 46 of whom had sons, and 29 of whom had no children. Clinical data of all women are given in Table 1.Table 1Clinical dataWomen with sonsWomen without childrenNumber of women4629Age (years)63 (29–93)59 (10–89)Cause of death:Infectious96Cancer145Cerebral86Vascular/myocardial1211Other3, i.e. amniotic fluid embolus, liver cirrhosis from alcohol abuse, cachexia1, i.e. liver failure from alcohol abuseBlood transfusion:Blood transfusion3015No blood transfusion1614Organs studied by ISH:13877Kidneys3120Livers3417Hearts4326Spleens3014 Open table in a new tab Archived paraffin-embedded tissues of the kidney, liver and heart from the autopsied cases were cut into 4 im sections, and deposited onto Superfrost plus glass slides (Menzel-Glaser, Germany). The sections were dried overnight at 37°C to improve tissue adherence. A Y-chromosome-specific DNA probe (23Lau YF Detection of Y-specific repeat sequences in normal and variant human chromosomes using in situ hybridization with bi- otinylated probes..Cytogenet Cell Genet. 1985; 39: 184-187Crossref PubMed Scopus (53) Google Scholar) was labeled with digoxigenin according to the standard Nick-translation protocol. After labeling, the probe was precipitated, dried and dissolved in a hybridization mixture (50% deionized formamide, 0.05 M sodium phosphate buffer pH 7.0, 2 × 0.3 mol/L NaCl, 30 mmol/L Na citrate [2 * SSC] and 10% dextran sulphate). Cot-1 DNA was added to the hybridization mixture. Slides were deparaffinized in xylene and dehydrated in an ethanol series followed by a distilled water rinse. The sections were pretreated with 0.05 M citrate buffer (pH 6.0) at 80°C for 80 min, rinsed in pre-warmed distilled water at 37°C, followed by a 0.5% Pepsin (Serva Electrophoresis GmbH, Heidelberg, Germany) in 0.01 M HCl at 37°C for 20 min, for enzyme digestion. Slides were then dehydrated in upgraded ethanol and air dried. Tissue sections on each slide were covered with a 30 µL hybridization mixture containing 5 ng/µL labeled probe. They were then denatured on a 80°C metal plate for 10 min and incubated at 37°C overnight. The next day, the sections were washed three times in 2 * SSC/0.1% Tween at 37°C and three times in 0.1 * SSC at 60°C. To visualize the Dig-labeled probe,sections were incubated consecutively with a mouse-anti-Dig monoclonal antibody (Sigma-Aldrich, St. Louis, MO), rabbit-anti-mouse immunoglobulin- HRP (Dako, Glostrup, Denmark) and swine-anti-rabbit immunoglobulin-HRP (Dako) at room temperature. Finally, the sections were developed with Nova Red Vector for 10 minutes. A hematoxylin staining served as a background. An X-chromosome-specific DNA probe (24Willard HF Smith KD Sutherland J Isolation and characterization of a major tandem repeat family from the human X chromosome..Nucleic Acids Res. 1983; 11: 2017-2033Crossref PubMed Scopus (230) Google Scholar) was hybridized according to the same method as described above. Tissue samples from either a male or a female Eurotransplant kidney that were rejected for transplantation because of technical reasons, served as positive controls for the in situ hybridization of the Y- and the X-chromosome, respectively. All slides were evaluated by at least two observers. A sample was scored positive if in one or more nuclei a red-brown-stained dot was present, with a similar size and staining intensity as those of the positive controls samples. The background was clear, and the stained dots were specifically present in the cell nuclei. Sporadically, a non-specific pattern of Nova Red Vector posi- tivity was found with numerous little speckles in the nucleus, or with a blurry staining pattern. The non-specifically stained cells seem to be leucocytes, possibly plasma cells, and were not counted positive. To confirm the results from the in situ hybridization, we performed a nested polymerase chain reaction (PCR). DNA was extracted from paraffin- embedded kidney specimens with the Wizard® Genomic DNA Purification Kit (Promega, Madison, WI, USA) from 7 women who bore sons. Two of these specimens were scored positive by in situ hybridization, and five specimens were scored negative. A specific Y-chromosome sequence was detected by amplifying DNA from a non-functional Sex-determining region Y (SRY) gene in a nested PCR with primers designed with the Beacon Designer (Bio-Rad laboratories, Hercules, CA, USA) software. The first amplification was done with primers Y1-1 that has the sequence 5′-CGC ATT CAT CGT GTG GTC TCG-3′ and Y1-2 that has the sequence 5′-lll TCG GCT TCA GTA AGC ATT TTC C-3′ (product of 120 bp). The nested amplification was done with primers Y1-3 that has the sequence 5′-TCA GAC GCG CAA GAT GGC TC-3′ and Y1-4 that has the sequence 5′-AGT AAG CAT TTT CCA CTG GTATCC C-3′ (product of 88 bp). Approximately, 20 ng DNA were used in a 25 µL assay containing 2.5 µL 10 × PCR buffer, 300 pmol of each primer, 0.5 µL of 10 mM of each dNTP and 0.1 U Amplitaq DNA Polymerase (Roche Applied Science, Indianapolis, IN, USA). The conditions for amplification were denaturation at 95°C for 30 s, annealing at 60°C for 30 s and extension at 72°C for 30 s for 35 cycles. Two microliter of this PCR product was used for a nested PCR, which was performed by 45 cycles of denaturation at 95°C for 30 s, annealing at 60°C for 30 s and extension at 72°C for 30 s. All PCR analyses contained a blank (water without DNA) for a negative control and a known positive sample for the Y-chromosome (male DNA). A PCR for a non-gender-related DNA fragment was used to verify the quality of the isolated DNA. To avoid contamination, all experiments were performed by a female technician. Pre-amplification steps were carried out in a separate room, in a safety hood that was sterilized each time by UV light. Furthermore, for the nested PCR, each sample was accompanied by an extra blank control. The resulting 88 bp Y-chromosome-specific fragment was identified by ethidium bromide staining after electrophoresis on a 3% agarose gel. The 88 bp PCR product was sequenced to confirm its identity. The fragment was sequenced with Y1-3 forward primer. The sequence results were verified for homology with the SRY gene using BLAST search against Genbank on the NCBI Website. Y-chromosome positive cells, as determined by in situ hy-bridization, were found in 13 kidneys, 10 livers, 4 hearts and 8 spleens. In Table 2, the division of the positive tissue samples among women with sons and women without children is presented, showing no signiicant difference between these two groups. In Table 3, the division of the positive tissue samples among women with and without blood transfusions is presented, also showing no significant difference between these two groups. Combining the data from the child and blood transfusion history, there were 30 women with a positive blood transfusion history with sons, 16 women with a negative blood transfusion history with sons, 15 women with a positive blood transfusion history without children and 14 women with a negative blood transfusions history without children. The number of women who had chimerism in any organ in these four groups, respectively, were: 9 of 30, 5 of 16, 7 of 15 and 6 of 14. We calculated the maximum number of male-nucleated cells present in the blood transfusion, and this number showed no significant relation with the number of positive tissue samples in any of the tested organ specimens (Figure 1). There was also no statistically significant relationship between the number of positive tissue specimens of any organ with either the child or the blood transfusion status.Table 2Results in situ hybridization for the Y-chromosomeWomen with sons (N = 46)Women without children (N = 29)Organs scored positive1619Women with at least one organ scored positive14/46 (30%)13/29 (44%)Age of women with at least one organ scored positive62 (29–93 yr)62 (29–89 yr)Women with more than one organ scored positive1 with liver and kidney 1 with liver and spleen1 with liver, kidney and spleen 1 with kidney, heart and spleen1 with liver and kidney 1 with liver and spleenKidneys scored positive6/31 (19%)7/20 (35%)Livers scored positive7/34 (21%)3/17 (18%)Hearts scored positive0/43 (0%)4/26 (15%)Spleens scored positive3/30 (10%)5/14 (36%) Open table in a new tab Table 3Blood transfusion dataY-chromosome positive cellsNo Y-chromosome positive cellsTotalWomen with sons (N = 46)Blood transfusion92130No blood transfusion51116Total143246Women without children (N = 29)Blood transfusion7815No blood transfusion6814Total131629All women (N = 75)Blood transfusion162945No blood transfusion111930Total274875 Open table in a new tab There were 6 women with more than one organ showing positive results in the in situ hybridization for the Y- chromosome, but it was never the case that all organs evaluated in one woman were positive (Table 2). Most positive samples only contained a small number of dots. However, in 2 patients the kidneys had an extensive number of nuclei in which the Y-chromosome was present. The results of each organ will be described separately below. In 13 of 51 kidney tissue samples, the Y-chromosome was detected. The positive cells were present in glomeruli and tubules. In two women, who had sons, up to 10% of the cells were positive ( Figure 2A). These women were 77 and 81 years old, and died of sepsis and a ruptured aneurysm of the vertebral artery, respectively. The 77 year old had received a blood transfusion, whereas the 81 year old had received no blood transfusion. From the 77 year old, also tissue specimens of liver, heart and spleen were incorporated in the study, but these were negative. From the 81 year old, a specimen from the heart was incorporated in the study, which was negative. In the other positive kidney samples, only one or two cells with Y-chromosomes were found (Figure 2B). Table 4 gives a division of positive kidney samples in relation to the cause of death, child status and previous blood transfusions.Table 4Women with a Y-chromosome identified by ISH in at least one organPatientAge at deathKidneyLiverHeartSpleenBlood transfusionCause of deathWomen with sons129+–––YesAdenocarcinoma of the lung238++––YesAmniotic fluid embolus343–+––YesCytomegalovirus pneumonia/sepsis444–––+NoSaddle embolus546+na–naYesCerebral hemorrhage657–––+NoNeuro‐endocrine carcinoma765na+na–YesSepsis867–+–+NoMyocardial infarction971–+––NoPneumonia1074–+––YesLeft ventricle fibrillation1176+na–naYesCardiac failure1277+*Extensive chimerism–––YesSepsis1381+*Extensive chimerismna–naNoRuptured aneurysm of the vertebral artery1493–+––YesCachexiaWomen without children1529na–+–YesMetastasized carcinoma of the ovary1638–+++YesMyocardial infarction1743+––+YesCerebral hemorrhage1845–+–naNoMyocardial infarction1952na+–naNoMetastasized melanoma2057––+–YesCytomegalovirus infection2166++–+NoLiver failure from alcohol abuse2268––+naNoMyocardial infarction2369na––6+YesSepsis2480na+–naYesPneumonia2581++––NoCerebral hemorrhage2686–+–naYesCerebral hemorrhage2789nana–+NoSepsisna = not available* Extensive chimerism Open table in a new tab na = not available In 10 of 51 liver tissue samples, the Y-chromosome was detected. The positive cells seemed to be present in hepa- tocytes (Figure 2C), and sometimes in the infiltrates of portal triads. Positive cells were scarce, up to 10 in each tissue sample. Table 4 gives a division of positive liver samples in relation to the cause of death, child status and previous blood transfusions. All tissue specimens from women with sons were negative (0/43). However, in the group of women without children, 4 of 26 heart specimens showed Y-chromosome positive cells, which appeared to be cardiomyocytes (Figure 2D). Table 4 gives a division of positive heart samples in relation to the cause of death, child status and previous blood transfusions. In 8 of 44 spleen tissue samples positive cells were found (Figure 2E). In all tissue samples, a number of cells were positive, but not more than 10. Aspecific staining occurred more in the spleen than in other organs. Table 4 gives a division of positive spleen samples in relation to the cause of death, child status and previous blood transfusions. Sequencing confirmed the identity of the Y-chromosome- specific product amplified from two DNA samples of women with sons, which were scored positive by in situ hybridization (ISH). The amplified product had homology with the SRY-sequence unique to the Y-chromosome, indicating that it was a male chromosome sequence sequence and not an irrelevant product. Y-chromosome-specific DNA was detected in bo" @default.
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• W2037355105 date "2005-06-01" @default.
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• W2037355105 title "Chimerism in Kidneys, Livers and Hearts of Normal Women: Implications for Transplantation Studies" @default.
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• W2037355105 doi "https://doi.org/10.1111/j.1600-6143.2005.00858.x" @default.
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