Molecular and Immunohistochemical Characterization of Historical Long-Term Preserved Fixed Tissues from Different Human Organs
Identifieur interne : 000135 ( Main/Merge ); précédent : 000134; suivant : 000136Molecular and Immunohistochemical Characterization of Historical Long-Term Preserved Fixed Tissues from Different Human Organs
Auteurs : Maja Hühns ; Paula Röpenack ; Andreas ErbersdoblerSource :
- PLoS ONE [ 1932-6203 ] ; 2015.
Abstract
University and museum collections are very important sources of biological samples that can be used to asses the past and present genetic diversity of many species. Modern genetic and immunohistochemical techniques can be used on long-term preserved fixed tissues from museum specimens to answer epidemiological questions. A proof of principle was established to apply modern molecular genetics and immunohistochemical methods to these old specimens and to verify the original diagnosis. We analysed 19 specimens from our university collection including human organs that had been in fixative for more than 80 years. The tissues originated from lung, colon, brain, heart, adrenal gland, uterus and skin. We isolated amplifiable DNA from these wet preparations and performed mutational analysis of BRAF, KRAS and EGFR. The tissues were also embedded in paraffin and used for modern histology and immunohistochemistry. Our data show that amplifiable DNA is extractable and ranged from 0.25 to 22.77 μg of total DNA. In three specimens BRAFV600E or KRASG12D mutations were found. Additionally, expression of different proteins like vimentin and GFAP was detected immunohistochemical in six investigated specimens. On the basis of our results the original diagnosis was altered in three specimens. Our work showed that it is possible to extract amplifiable DNA suitable for sequence analysis from long-term fixed tissue. Furthermore, histology and immunohistochemistry is feasible in specimens fixed long time ago. We conclude that these old preparations are suitable for further epidemiological research and that our methods open up new opportunities for future studies.
Url:
DOI: 10.1371/journal.pone.0135297
PubMed: 26252375
PubMed Central: 4529272
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Molecular and Immunohistochemical Characterization of Historical Long-Term Preserved Fixed Tissues from Different Human Organs</title><author><name sortKey="Huhns, Maja" sort="Huhns, Maja" uniqKey="Huhns M" first="Maja" last="Hühns">Maja Hühns</name><affiliation><nlm:aff id="aff001"></nlm:aff></affiliation></author><author><name sortKey="Ropenack, Paula" sort="Ropenack, Paula" uniqKey="Ropenack P" first="Paula" last="Röpenack">Paula Röpenack</name><affiliation><nlm:aff id="aff001"></nlm:aff></affiliation></author><author><name sortKey="Erbersdobler, Andreas" sort="Erbersdobler, Andreas" uniqKey="Erbersdobler A" first="Andreas" last="Erbersdobler">Andreas Erbersdobler</name><affiliation><nlm:aff id="aff001"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">26252375</idno><idno type="pmc">4529272</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4529272</idno><idno type="RBID">PMC:4529272</idno><idno type="doi">10.1371/journal.pone.0135297</idno><date when="2015">2015</date><idno type="wicri:Area/Pmc/Corpus">000025</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000025</idno><idno type="wicri:Area/Pmc/Curation">000025</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000025</idno><idno type="wicri:Area/Pmc/Checkpoint">000047</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000047</idno><idno type="wicri:Area/Ncbi/Merge">000E96</idno><idno type="wicri:Area/Ncbi/Curation">000E96</idno><idno type="wicri:Area/Ncbi/Checkpoint">000E96</idno><idno type="wicri:Area/Main/Merge">000135</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Molecular and Immunohistochemical Characterization of Historical Long-Term Preserved Fixed Tissues from Different Human Organs</title><author><name sortKey="Huhns, Maja" sort="Huhns, Maja" uniqKey="Huhns M" first="Maja" last="Hühns">Maja Hühns</name><affiliation><nlm:aff id="aff001"></nlm:aff></affiliation></author><author><name sortKey="Ropenack, Paula" sort="Ropenack, Paula" uniqKey="Ropenack P" first="Paula" last="Röpenack">Paula Röpenack</name><affiliation><nlm:aff id="aff001"></nlm:aff></affiliation></author><author><name sortKey="Erbersdobler, Andreas" sort="Erbersdobler, Andreas" uniqKey="Erbersdobler A" first="Andreas" last="Erbersdobler">Andreas Erbersdobler</name><affiliation><nlm:aff id="aff001"></nlm:aff></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>University and museum collections are very important sources of biological samples that can be used to asses the past and present genetic diversity of many species. Modern genetic and immunohistochemical techniques can be used on long-term preserved fixed tissues from museum specimens to answer epidemiological questions. A proof of principle was established to apply modern molecular genetics and immunohistochemical methods to these old specimens and to verify the original diagnosis. We analysed 19 specimens from our university collection including human organs that had been in fixative for more than 80 years. The tissues originated from lung, colon, brain, heart, adrenal gland, uterus and skin. We isolated amplifiable DNA from these wet preparations and performed mutational analysis of BRAF, KRAS and EGFR. The tissues were also embedded in paraffin and used for modern histology and immunohistochemistry. Our data show that amplifiable DNA is extractable and ranged from 0.25 to 22.77 μg of total DNA. In three specimens BRAF<sup>V600E</sup> or KRAS<sup>G12D</sup> mutations were found. Additionally, expression of different proteins like vimentin and GFAP was detected immunohistochemical in six investigated specimens. On the basis of our results the original diagnosis was altered in three specimens. Our work showed that it is possible to extract amplifiable DNA suitable for sequence analysis from long-term fixed tissue. Furthermore, histology and immunohistochemistry is feasible in specimens fixed long time ago. We conclude that these old preparations are suitable for further epidemiological research and that our methods open up new opportunities for future studies.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Anastasiou, E" uniqKey="Anastasiou E">E Anastasiou</name></author><author><name sortKey="Mitchell, Pd" uniqKey="Mitchell P">PD Mitchell</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="De Melo, Fl" uniqKey="De Melo F">FL de Melo</name></author><author><name sortKey="De Mello, Jc" uniqKey="De Mello J">JC de Mello</name></author><author><name sortKey="Moreira, Fam" uniqKey="Moreira F">FAM Moreira</name></author><author><name sortKey="Nunes, K" uniqKey="Nunes K">K Nunes</name></author><author><name sortKey="Eggers, S" uniqKey="Eggers S">S Eggers</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Graeber, Mb" uniqKey="Graeber M">MB Graeber</name></author><author><name sortKey="Kosel, S" uniqKey="Kosel S">S Kösel</name></author><author><name sortKey="Grasbon Frodl, E" uniqKey="Grasbon Frodl E">E Grasbon-Frodl</name></author><author><name sortKey="Moller, Hj" uniqKey="Moller H">HJ Möller</name></author><author><name sortKey="Mehraein, P" uniqKey="Mehraein P">P Mehraein</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Moller, Hj" uniqKey="Moller H">HJ Möller</name></author><author><name sortKey="Graeber, Mb" uniqKey="Graeber M">MB Graeber</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rocken, C" uniqKey="Rocken C">C Röcken</name></author><author><name sortKey="Widulin, N" uniqKey="Widulin N">N Widulin</name></author><author><name sortKey="Schnalke, T" uniqKey="Schnalke T">T Schnalke</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fornaciari, G" uniqKey="Fornaciari G">G Fornaciari</name></author><author><name sortKey="Giuffra, V" uniqKey="Giuffra V">V Giuffra</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Worobey, M" uniqKey="Worobey M">M Worobey</name></author><author><name sortKey="Gemmel, M" uniqKey="Gemmel M">M Gemmel</name></author><author><name sortKey="Teuwen, De" uniqKey="Teuwen D">DE Teuwen</name></author><author><name sortKey="Haselkorn, T" uniqKey="Haselkorn T">T Haselkorn</name></author><author><name sortKey="Kunstman, K" uniqKey="Kunstman K">K Kunstman</name></author><author><name sortKey="Bunce, M" uniqKey="Bunce M">M Bunce</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Westermark, P" uniqKey="Westermark P">P Westermark</name></author><author><name sortKey="Nilsson, Gt" uniqKey="Nilsson G">GT Nilsson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fox, H" uniqKey="Fox H">H Fox</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tsutsumi, Y" uniqKey="Tsutsumi Y">Y Tsutsumi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Blum, F" uniqKey="Blum F">F Blum</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Coombs, N" uniqKey="Coombs N">N Coombs</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Duval, K" uniqKey="Duval K">K Duval</name></author><author><name sortKey="Aubin, Ra" uniqKey="Aubin R">RA Aubin</name></author><author><name sortKey="Elliott, J" uniqKey="Elliott J">J Elliott</name></author><author><name sortKey="Gorn Hondermann, I" uniqKey="Gorn Hondermann I">I Gorn-Hondermann</name></author><author><name sortKey="Birnboim, Hc" uniqKey="Birnboim H">HC Birnboim</name></author><author><name sortKey="Jonker, D" uniqKey="Jonker D">D Jonker</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gilbert, M" uniqKey="Gilbert M">M Gilbert</name></author><author><name sortKey="Thomas, P" uniqKey="Thomas P">P Thomas</name></author><author><name sortKey="Haselkorn, T" uniqKey="Haselkorn T">T Haselkorn</name></author><author><name sortKey="Bunce, M" uniqKey="Bunce M">M Bunce</name></author><author><name sortKey="Sanchez, Jj" uniqKey="Sanchez J">JJ Sanchez</name></author><author><name sortKey="Lucas, Sb" uniqKey="Lucas S">SB Lucas</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Okello, Jb" uniqKey="Okello J">JB Okello</name></author><author><name sortKey="Zurek, J" uniqKey="Zurek J">J Zurek</name></author><author><name sortKey="Devault, Am" uniqKey="Devault A">AM Devault</name></author><author><name sortKey="Kuch, M" uniqKey="Kuch M">M Kuch</name></author><author><name sortKey="Okwi, A" uniqKey="Okwi A">A Okwi</name></author><author><name sortKey="Sewankambo, Nk" uniqKey="Sewankambo N">NK Sewankambo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Santos, Mc" uniqKey="Santos M">MC Santos</name></author><author><name sortKey="Saito, Cp" uniqKey="Saito C">CP Saito</name></author><author><name sortKey="Line, Sr" uniqKey="Line S">SR Line</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shi, S" uniqKey="Shi S">S Shi</name></author><author><name sortKey="Datar, R" uniqKey="Datar R">R Datar</name></author><author><name sortKey="Liu, C" uniqKey="Liu C">C Liu</name></author><author><name sortKey="Wu, L" uniqKey="Wu L">L Wu</name></author><author><name sortKey="Zhang, Z" uniqKey="Zhang Z">Z Zhang</name></author><author><name sortKey="Richard, J" uniqKey="Richard J">J Richard</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Santos, S" uniqKey="Santos S">S Santos</name></author><author><name sortKey="Sa, D" uniqKey="Sa D">D Sá</name></author><author><name sortKey="Bastos, E" uniqKey="Bastos E">E Bastos</name></author><author><name sortKey="Guedes Pinto, H" uniqKey="Guedes Pinto H">H Guedes-Pinto</name></author><author><name sortKey="Gut, I" uniqKey="Gut I">I Gut</name></author><author><name sortKey="G Rtner, F" uniqKey="G Rtner F">F Gärtner</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Paireder, S" uniqKey="Paireder S">S Paireder</name></author><author><name sortKey="Werner, B" uniqKey="Werner B">B Werner</name></author><author><name sortKey="Bailer, J" uniqKey="Bailer J">J Bailer</name></author><author><name sortKey="Werther, W" uniqKey="Werther W">W Werther</name></author><author><name sortKey="Schmid, E" uniqKey="Schmid E">E Schmid</name></author><author><name sortKey="Patzak, B" uniqKey="Patzak B">B Patzak</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Remmele, W" uniqKey="Remmele W">W Remmele</name></author><author><name sortKey="Stegner, He" uniqKey="Stegner H">HE Stegner</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sambrook, J" uniqKey="Sambrook J">J Sambrook</name></author><author><name sortKey="Fritsch, Ef" uniqKey="Fritsch E">EF Fritsch</name></author><author><name sortKey="Maniatis, T" uniqKey="Maniatis T">T Maniatis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Karlsen, F" uniqKey="Karlsen F">F Karlsen</name></author><author><name sortKey="Kalantari, M" uniqKey="Kalantari M">M Kalantari</name></author><author><name sortKey="Chitemerere, M" uniqKey="Chitemerere M">M Chitemerere</name></author><author><name sortKey="Johansson, B" uniqKey="Johansson B">B Johansson</name></author><author><name sortKey="Hagmar, B" uniqKey="Hagmar B">B Hagmar</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Akalu, A" uniqKey="Akalu A">A Akalu</name></author><author><name sortKey="Reichardt, Jk" uniqKey="Reichardt J">JK Reichardt</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ben Ezra, J" uniqKey="Ben Ezra J">J Ben-Ezra</name></author><author><name sortKey="Johnson, Da" uniqKey="Johnson D">DA Johnson</name></author><author><name sortKey="Rossi, J" uniqKey="Rossi J">J Rossi</name></author><author><name sortKey="Cook, N" uniqKey="Cook N">N Cook</name></author><author><name sortKey="Wu, A" uniqKey="Wu A">A Wu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jackson, Dp" uniqKey="Jackson D">DP Jackson</name></author><author><name sortKey="Lewis, Fa" uniqKey="Lewis F">FA Lewis</name></author><author><name sortKey="Taylor, Gr" uniqKey="Taylor G">GR Taylor</name></author><author><name sortKey="Boylston, Aw" uniqKey="Boylston A">AW Boylston</name></author><author><name sortKey="Quirke, P" uniqKey="Quirke P">P Quirke</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chaw, Y" uniqKey="Chaw Y">Y Chaw</name></author><author><name sortKey="Fen, M" uniqKey="Fen M">M Fen</name></author><author><name sortKey="Crane, Le" uniqKey="Crane L">LE Crane</name></author><author><name sortKey="Lange, P" uniqKey="Lange P">P Lange</name></author><author><name sortKey="Shapiro, R" uniqKey="Shapiro R">R Shapiro</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Feldman, M" uniqKey="Feldman M">M Feldman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yasui, M" uniqKey="Yasui M">M Yasui</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ferrer, I" uniqKey="Ferrer I">I Ferrer</name></author><author><name sortKey="Armstrong, J" uniqKey="Armstrong J">J Armstrong</name></author><author><name sortKey="Capellari, S" uniqKey="Capellari S">S Capellari</name></author><author><name sortKey="Parchi, P" uniqKey="Parchi P">P Parchi</name></author><author><name sortKey="Arzberger, T" uniqKey="Arzberger T">T Arzberger</name></author><author><name sortKey="Bell, J" uniqKey="Bell J">J Bell</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lyck, L" uniqKey="Lyck L">L Lyck</name></author><author><name sortKey="Dalmau, I" uniqKey="Dalmau I">I Dalmau</name></author><author><name sortKey="Chemnitz, J" uniqKey="Chemnitz J">J Chemnitz</name></author><author><name sortKey="Finsen, B" uniqKey="Finsen B">B Finsen</name></author><author><name sortKey="Schr Der, Hd" uniqKey="Schr Der H">HD Schrøder</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Miller, Ma" uniqKey="Miller M">MA Miller</name></author><author><name sortKey="Ramos Vara, Ja" uniqKey="Ramos Vara J">JA Ramos-Vara</name></author><author><name sortKey="Kleiboeker, Sb" uniqKey="Kleiboeker S">SB Kleiboeker</name></author><author><name sortKey="Larson, Rl" uniqKey="Larson R">RL Larson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Webster, Jd" uniqKey="Webster J">JD Webster</name></author><author><name sortKey="Miller, Ma" uniqKey="Miller M">MA Miller</name></author><author><name sortKey="Dusold, D" uniqKey="Dusold D">D Dusold</name></author><author><name sortKey="Ramos Vara, Ja" uniqKey="Ramos Vara J">JA Ramos-Vara</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Werner, M" uniqKey="Werner M">M Werner</name></author><author><name sortKey="Chott, A" uniqKey="Chott A">A Chott</name></author><author><name sortKey="Fabiano, A" uniqKey="Fabiano A">A Fabiano</name></author><author><name sortKey="Battifora, H" uniqKey="Battifora H">H Battifora</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wasielewski, R" uniqKey="Wasielewski R">R Wasielewski</name></author><author><name sortKey="Mengel, M" uniqKey="Mengel M">M Mengel</name></author><author><name sortKey="Nolte, M" uniqKey="Nolte M">M Nolte</name></author><author><name sortKey="Werner, M" uniqKey="Werner M">M Werner</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Petrov, D" uniqKey="Petrov D">D Petrov</name></author><author><name sortKey="Vlassov, V" uniqKey="Vlassov V">V Vlassov</name></author><author><name sortKey="Kalaydjiev, G" uniqKey="Kalaydjiev G">G Kalaydjiev</name></author><author><name sortKey="Plochev, Ma" uniqKey="Plochev M">MA Plochev</name></author><author><name sortKey="Obretenov, Ed" uniqKey="Obretenov E">ED Obretenov</name></author><author><name sortKey="Stanoev, Vi" uniqKey="Stanoev V">VI Stanoev</name></author></analytic></biblStruct></listBibl></div1></back></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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