Development of A Novel Therapy for Neonatal Intractable Disease Using Autologous Amniotic Fluid Stem Cell Sheets

• Project/Area Number: 16K15536
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Embryonic/Neonatal medicine
• Research Institution: Keio University
• Principal Investigator: Mamoru Tanaka 慶應義塾大学, 医学部(信濃町), 教授 (20207145)
• Co-Investigator(Kenkyū-buntansha): 落合 大吾 慶應義塾大学, 医学部(信濃町), 講師 (80348713)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2018: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 羊水 / 幹細胞 / 創傷治癒 / 線維化 / 細胞シート / 再生医療 / 抗繊維化作用 / 新生児

Outline of Final Research Achievements

Fetal wounds before 24 weeks of gestation are known to heal without fibrotic scarring, and this characteristic is preserved beyond animal species. Therefore, we hypothesized that human amniotic fluid stem cells (hAFS) have more potential for anti-fibrosis than other mesenchymal stem cells. In addition, hAFS that have pre-differentiated into target cells, such as neurons and cardiomyocytes, could be used to create hAFS sheets.
In this study, we demonstrated that hAFS are multipotent and could be differentiated into neuronal cells and cardiomyocyte-like cells. Additionally, we successfully produced a cell sheet using hAFS and confirmed its formation and strong anti- fibrotic potential using a murine model. However, we were unable to create a pre-differentiated hAFS sheets.

Academic Significance and Societal Importance of the Research Achievements

細胞工学技術の進歩により細胞シートを作成することが可能になった。細胞シートは、成人疾患には臨床応用が開始されているが、 新生児疾患ではほとんど検討されてこなかった。新生児にヒト羊水幹細胞（human amniotic fluid stem cells: hAFS）を用いた細胞治療を行う際、「頻回の細胞注射で生着する幹細胞数を増やす」方法が考えられる。しかし、新生児には負担が強いため、非侵襲的に幹細胞生着率を上げるツールとしてhAFS細胞シートの開発を試みた。我々は、hAFS細胞シート作成に成功し、その細胞シートは強力な抗線維化作用を持つことを動物実験で明らかにした。

Research Products

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• [Journal Article] Retrospective multicenter study of leaving the placenta in situ for patients with placenta previa on a cesarean scar. (2018)
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• [Journal Article] 羊水幹細胞を用いた胎児治療（現状と将来像） (2018)
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• [Journal Article] 多胎妊娠と糖代謝異常より良い管理を目指して 双胎妊娠における妊娠糖尿病の糖代謝異常重症度に関する検討 (2018)
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• [Journal Article] 妊娠糖尿病管理中の低カリウム性四肢麻痺を契機に検出されたCushing症候群合併妊娠の1例 (2018)
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• [Journal Article] Discordant fetal phenotype of hypophosphatasia in two siblings. (2018)
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• [Journal Article] ZEB1 expression is a potential indicator of invasive endometriosis. (2017)
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• [Journal Article] Mid-trimester residual cervical length and the risk of preterm birth in pregnancies after abdominal radical trachelectomy: a retrospective analysis. (2017)
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• [Journal Article] Association of common polymorphisms with gestational diabetes mellitus in Japanese women: A case-control study. (2017)
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• [Journal Article] Low dose resveratrol ameliorates mitochondrial respiratory dysfunction and enhances cellular reprogramming. (2017)
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• [Journal Article] H1foo Has a Pivotal Role in Qualifying Induced Pluripotent Stem Cells (2016)
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  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] A POTENTIAL THERAPY FOR FETAL MYELOMENINGOCELE BY HUMAN AMNIOTIC FLUID STEM CELLS. (2018)
  • Author(s): Abe Y, Ochiai D, Masuda H, Sato Y, Otani T, Fukutake M, Tanaka M
  • Organizer: International Society for Stem Cell Research (ISSCR 2018)
  • Int'l Joint Research
• [Presentation] The therapeutic potential of human amniotic fluid stem cells on the chronic phase of neonatal hypoxic-ischemic encephalopathy in mice (2018)
  • Author(s): Otani T, Ochiai D, Masuda H, Fukutake M, Sato Y, Ikenoue S, Miyakoshi K, Tanaka M
  • Organizer: 8th Korea-Taiwan-Japan Symposium on Maternal-Fetal Medicine
  • Int'l Joint Research
• [Presentation] THE THERAPEUTIC EFFECTS OF HUMAN AMNIOTIC FLUID STEM CELLS ON THE CHRONIC PHASE OF NEONATAL HYPOXIC-ISCHEMIC ENCEPHALOPATHY (2018)
  • Author(s): Otani T, Ochiai D, Masuda H, Fukutake M, Sato Y, Ikenoue S, Miyakoshi K, Tanaka M
  • Organizer: 20th Congress of the Federation of Asia and Oceania Perinatal Societies (FAOPS 2018)
  • Int'l Joint Research
• [Presentation] A NOVEL STRATEGY TO TREAT FETAL MYELOMENINGOCELE USING HUMAN AMNIOTIC FLUID STEM CELLS (2018)
  • Author(s): Sato Y, Ochiai D, Abe Y, OtaniT, Fukutake M, Ikenoue S, Masuda H, Miyakoshi K, Tanaka M
  • Organizer: 20th Congress of the Federation of Asia and Oceania Perinatal Societies (FAOPS 2018)
  • Int'l Joint Research
• [Presentation] Pregnancy Outcomes After Abdominal Radical Trachelectomy for Early-Stage Cervical Cancer (2018)
  • Author(s): Ochiai D, Tanaka M
  • Organizer: 2018 Kangwon-Keio International symposium
  • Int'l Joint Research / Invited
• [Presentation] Human Amniotic Fluid Stem Cells: Therapeutic Potential for Perinatal Patients with Intractable Neurological Disease (2018)
  • Author(s): Sato Y, Tanaka M
  • Organizer: 2018 Kangwon-Keio International symposium
  • Int'l Joint Research / Invited
• [Presentation] ラット胎仔脊髄髄膜瘤モデルを使用したヒト羊水幹細胞の羊水腔内注入による胎児治療効果について (2018)
  • Author(s): 阿部雄志, 落合大吾, 大谷利光, 福武麻里絵, 佐藤 佑, 秋葉洋平, 松本 直, 升田博隆, 宮越 敬, 田中 守, 青木大輔
  • Organizer: 第70回 日本産科婦人科学会
• [Presentation] 新生仔低酸素虚血性脳症モデルマウスに対するヒト羊水幹細胞鼻腔内投与の治療効果に関する検討 (2018)
  • Author(s): 大谷利光, 落合大吾, 阿部雄志, 福武 麻里絵, 佐藤 佑, 秋葉洋平, 松本 直, 升田博隆, 宮越 敬, 田中 守, 青木大輔
  • Organizer: 第70回 日本産科婦人科学会
• [Presentation] 新生仔低酸素虚血性脳症モデルマウスに対するヒト羊水幹細胞投与の治療効果に関する検討 (2018)
  • Author(s): 大谷利光, 落合大吾, 佐藤 佑, 福武 麻里絵, 松本 直, 宮越 敬, 田中 守
  • Organizer: 第54回 日本周産期・新生児医学会
• [Presentation] A novel strategy to treat fetal myelomeningocele using human amniotic fluid stem cells (2018)
  • Author(s): Yushi Abe, Daigo Ochiai, Hirotaka Masuda, Toshimitsu Otani, Marie Fukutake, Youhei Akiba, Tadashi Matsumoto, Kei Miyakoshi, Mamoru Tanaka
  • Organizer: Society for Reproductive Investigation
  • Int'l Joint Research
• [Presentation] Human amniotic fluid stem cells have a unique potential to accelerate cutaneous wound healing with less fibrotic scarring like fetal wound healing (2018)
  • Author(s): Yu Sato, Daigo Ochiai, Hirotaka Masuda, Yushi Abe, Toshimitsu Otani, Marie Fukutake, Youhei Akiba, Tadashi Matsumoto, Kei Miyakoshi, Mamoru Tanaka
  • Organizer: Society for Reproductive Investigation
  • Int'l Joint Research
• [Presentation] Human amniotic fluid stem cells :A novel treatment option for neonatal cerebral palsy (2018)
  • Author(s): Toshimitsu Otani, Daigo Ochiai, Marie Fukutake, Yushi Abe, Yu Sato, Youhei Akiba, Tadashi Matsumoto, Hirotaka Masuda, Kei Miyakoshi, Mamoru Tanaka
  • Organizer: Society for Reproductive Investigation
  • Int'l Joint Research
• [Presentation] ヒト羊水幹細胞の投与前調整法に関する検討 (2017)
  • Author(s): 阿部雄志，落合大吾，福武麻里絵，大谷利光，升田博隆，秋葉洋平，松本直，宮越敬，田中守
  • Organizer: 臍帯血による再生医療研究会
• [Presentation] ヒト羊水幹細胞の羊水腔内注入によるラット脊髄髄膜瘤の治療効果に関する検討 (2017)
  • Author(s): 落合大吾, 阿部雄志, 大谷利光, 福武麻里絵, 升田博隆, 佐藤佑, 秋葉洋平, 松本直, 宮越敬, 田中守
  • Organizer: 日本胎児治療学会
• [Presentation] The effect of human amniotic fluid stem cells on the cutaneous wound healing (2016)
  • Author(s): Daigo Ochiai, Marie Fukutake, Hirotaka Masuda, Tadashi Matsumoto, Kei Miyakoshi, and Mamoru Tanaka
  • Organizer: 19th Congress of the Federation of Asia and Oceania Perinatal Societies
  • Place of Presentation: 台北（台湾）
  • Year and Date: 2016-12-01
  • Int'l Joint Research
• [Presentation] ヒト羊水幹細胞がマウス皮膚創傷治癒に与える影響 (2016)
  • Author(s): 福武麻里絵、落合大吾、升田博隆、大谷利光、秋葉洋平、春日義史、池ノ上学、松本直、宮越敬、田中守
  • Organizer: 第14回 日本胎児治療学会学術集会
  • Place of Presentation: 聖隷浜松病院（静岡県浜松市）
  • Year and Date: 2016-11-18
• [Presentation] ヒト羊水幹細胞はマウス皮膚創傷治癒を促進する (2016)
  • Author(s): 福武麻里絵、落合大吾、大谷利光、秋葉洋平、春日義史、池ノ上学、松本直、宮越敬、田中守
  • Organizer: 第52回 日本周産期・新生児医学会学術集会
  • Place of Presentation: 富山国際会議場（富山県富山市）
  • Year and Date: 2016-07-16
• [Presentation] Conditioned medium from human amniotic fluid stem cell ameliorates glutamate-induced apoptosis (2016)
  • Author(s): 福武麻里絵、落合大吾、大谷利光、秋葉洋平、春日義史、池ノ上学、松本直、宮越敬、田中守、青木大輔
  • Organizer: 第68回 日本産科婦人科学会学術講演会
  • Place of Presentation: 東京国際フォーラム（東京都千代田区）
  • Year and Date: 2016-04-21