2014年4月10日木曜日

STAP細胞の特許書類における不適切な画像流用まとめ

PDFファイル: 2012年4月の特許出願書類(US 61/637,631 24.04.2012 (Pr. Doc.))
Document details: Country/Office: US, Number: 61/637,631, Filing date: 24 April 2012 (24.04.2012)
(この特許出願書類は、STAP細胞の特許(WO2013163296: GENERATING PLURIPOTENT CELLS DE NOVO)に関するWIPOのサイトDocumentのタブのページに掲載されている。)


小保方晴子氏の博士論文のChapter 3 (Fig.11とFig.14を含む)



不適切な画像流用1
小保方晴子氏の博士論文(2011年2月)のFig.11の骨髄sphere(機械的ストレス誘導)由来のin vitro分化画像と、2012年4月の特許出願書類のACCs(弱酸刺激誘導細胞)由来のin vitro分化画像(Fig.9A上段)が、同一で、不適切な流用です。



Animal Callus Cells (ACCs)のcallus は、”《植物》カルス、癒傷組織” にネーミングを由来しているようです([00127]や[00132] にplant callus について言及してあります)。Fig.7cには色々な臓器由来なACCsが示されてあります。つまり、ACCsはSTAPの別名と考えればよさそうです。
また、[00137] には下記ののようにACCs のネーミングの由来が記載されています。
[00137] Stress treated CD45 positive cells were cultured in B27-LIF medium, and within 5 days, GFP expressing spherical colonies were observed while no GFP expressing colonies were observed in the untreated control (Figure 1A). Sphereical colonies grew to approximately 70μm in diameter over the first 7 days, and spherical colonies could be maintained for another 7 days in that culture condition. The configuration of the colonies was slightly baroque, appearing more similar in shape to the callus seen in botany, rather than spheres. A cell colony generated by stress treatment was therefore referred to as an Animal Callus (AC). Cultured cells were dissociated and population analysis was then performed using FACS. The analysis revealed that the application of certain significant stimuli resulted in the generation of stress altered cells, now referred to as Animal Callus Cells (ACCs), that did not previously exist in the CD45 positive cell populations (Figure 1B).  ・・・ (略)

特許出願書類のFig.9(A)については書類の本文中において、下記のように説明されています。Fig.9(A)に用いられたACCsは、[00142]において、CD45 positive lymphocytes 由来と記載されています(CD45 positive lymphocytesがどの臓器由来かについては何も記載はないようです)。
[0030] Figures 9A-9E depict differentiaion of ACCs. Figure 9A depicts an in vitro differentiation assay. ACCs were sorted and plated into differentiation medium. ACCs differentiated into cells from three germ layers, and expressed ectoderm specific marker βIII tubulin and GFAP, mesenchymal marker, α-smooth muscle actin, and endoderm marker, α-fetoprotein and cytokeratin 7. (以下略) 
[00142] To assess the stemness of ACCs, their self-renewal potency and their differentiation potency were examined. To study their self-renewal potency, ACCs colonies derived from previously mature CD45 positive lymphocytes were dissociated into single cells, and plated into 96 well plates, with one cell per well in an effort to generate clonally derived populations. Ten days after plating, spherical colonies were seen in 12-16h and others divided in 30-34h. ACCs were passaged at least 5 times, with continued expression of Oct4 observed. Consequently, ACCs demonstrated a potential for self-renewal, and the potential to differentiate into cells from all three germ layers in vitro
[00143] ACs derived from mature GOF lymphocytes were again dissociated into single cells, sorted to contain only a population of cells that expressed GFP and then cultured in differentiation media. At 14-21 days after plating, cells expressed the ectoderm marker, βIII-tubulin and GFAP, the mesoderm marker, α-smooth muscle actin, and the endoderm marker, αfetoprotein and Cytokeratin 7 (Figure 9A). Thus, ACCs differentiated into cells representative of the three germ layers in vitro.00135
また、[00135]には、"Consequently, exposure to low pH was focused upon as the stress treatment of choice for the remainder of the study." という文章があることから、Fig.9(A)に用いられたACCsは、弱酸刺激(low pH)で誘導されたものであると考えれます。



不適切な画像流用

特許出願書類のFig.9(A)については書類の本文中の[00148] において、下記のように説明されています。
[00148] Developmental potential of ACCs. Finally, it was assesed whether ACCs possessed a developmental potential similar to that of plant callus cells. As an initial test for developmental potency, ACCs implanted subcutaneously in immunodeficienct (SCID) mice were studied. Six wees after transplantation, ACCs generated tissues representing all three germ layers (Figure 9B)

小保方晴子氏の博士論文Fig.14からNature誌の論文に不適切に流用されていたテラトーマ画像が、2012年4月24日出願の特許書類のFig.9Bにも異なる実験画像として不適切に流用されていたことが発覚

 Nature論文とは異なり特許出願書類では、蛍光写真(下段)だけでなく、H&E染色画像(上段)も流用しています。特許で、H&Eと蛍光の二つを丸ごと博士論文から流用しておいて、その後のNature誌論文では下段の蛍光写真だけを切り取って流用しています。取違えミスとは到底思えません。

 Nature誌の論文の方では、博士論文や特許書類の別のテラトーマ画像の下段だけを切り取り、元の上段のH&E画像は取り除いて、そこに、どこからか持ってきた不自然なH&E染色画像を、組み合わせているのです。


2 件のコメント:

  1. > STAP細胞の特許

    の、以下、基本情報。

    Pub. No.: WO/2013/163296
    International Application No.: PCT/US2013/037996
    International Filing Date: 24.04.2013
    Publication Date: 31.10.2013
    Applicants:
    THE BRIGHAM AND WOMEN'S HOSPITAL, INC. [US/US]
    RIKEN [JP/JP]
    TOKYO WOMEN'S MEDICAL UNIVERSITY [JP/JP]

    Inventors:
    VACANTI, Charles A.; (US).
    VACANTI, Martin P.; (US).
    KOJIMA, Koji; (US).
    OBOKATA, Haruko; (JP).
    WAKAYAMA, Teruhiko; (JP).
    SASAI, Yoshiki; (JP).
    YAMATO, Masayuki; (JP)

    返信削除
  2. 東京女子医科大学は何もしないのか?

    返信削除