⬺ศ໬⬡⫫⣽⬊(DFAT)䛻䜘䜛෌⏕་⒪Regenerative medicine using dedifferentiated fat (DFAT) cells᪥ᮏ኱Ꮫ་Ꮫ㒊 ᶵ⬟ᙧែᏛ⣔⣽⬊෌⏕䞉⛣᳜་Ꮫศ㔝ᩍᤵ㻌 ᯇᮏ㻌 ኴ㑻㻌
Taro Matsumoto, Department of Functional Morphology, Division of Cell Regeneration and Transplantation, Nihon University School of Medicine஦ᴗ䝥䝍 360i
60i JAPANN஦ᴗ䝥䝻䝰䞊䝍䞊䠖360ip┠ⓗ䞉⫼ᬒ䠄Purpose and Background䠅㻌 㛫ⴥ⣔ᖿ⣽⬊䠄㻹㻿㻯䠅䛿ᝈ⪅⮬㌟䛾య䛛䜙ㄪ〇䛷䛝䚸⭘⒆ཎᛶ䛜䛺䛟⛣᳜Ᏻ඲ᛶ䛜㧗䛔䛯䜑䚸ከ䛟䛾⮫ᗋ◊✲䛜⾜䜟䜜䛶䛔䜛䚹୍᪉䚸㻹㻿㻯䛿ᇵ㣴ᚋ䜒䝦䝔䝻䛺⣽⬊㞟ᅋ䛷䚸䛭䛾ရ㉁䛿ಶయᕪ䛜኱䛝䛟䚸䝗䝘䞊ᖺ㱋䜔ᇶ♏⑌ᝈ䛻ᙳ㡪䛥䜜䜛䛸䛔䛳䛯ၥ㢟Ⅼ䛜䛒䜛䚹෌⏕་⒪䜢୓ே䛻㐺ᛂ䛷䛝䜛୍⯡ⓗ䛺἞⒪䛻䛩䜛䛯䜑䛻䛿䚸䜘䜚Ᏻᐃ䛧䛯ရ㉁䛾ከ⬟ᛶ⣽⬊䜢ప䝁䝇䝖䛻ㄪ〇䛩䜛ᢏ⾡䛜ᮃ䜎䜜䜛䚹㻌㻌㻌⬺ศ໬⬡⫫⣽⬊䠄㻰㼑㼐㼕㼒㼒㼑㼞㼑㼚㼠㼕㼍㼠㼑㼐㻌 㼒㼍㼠㻌 㼏㼑㼘㼘䠖㻰㻲㻭㼀䠅䛿ᡂ⇍⬡⫫⣽⬊䜢ኳ஭ᇵ㣴䛸䛔䛖᪉ἲ䛷⬺ศ໬䛥䛫䜛䛣䛸䛻䜘䜚ᚓ䜙䜜䜛㻹㻿㻯䛸ྠ➼䛾ከ⬟ᛶ䜢᭷䛩䜛⣽⬊䛷䛒䜛䚹㻰㻲㻭㼀䛿ᑡ㔞䛾⬡⫫⤌⧊䛛䜙ᆒ㉁䛺㻹㻿㻯ᵝ⣽⬊䜢኱㔞ㄪ〇䛷䛝䜛ᢏ⾡䛷䛒䜛䛯䜑䚸ᐇ⏝ᛶ䛾㧗䛔἞⒪⏝⣽⬊䝋䞊䝇䛸䛧䛶ᮇᚅ䛷䛝䜛䚹㻌䛆㻌 ἞⒪⏝⣽⬊䛸䛧䛶䛾㻰㻲㻭㼀䛾฼Ⅼ㻌 䛇㻌㻰㻲㻭㼀㻌ᡂ⇍⬡⫫⣽⬊㻌ᑡ㔞䛾⬡⫫⤌⧊䛛䜙㧗ຠ⋡䛻኱㔞ㄪ〇䛷䛝䜛㻌z⬡⫫⣽⬊䛛䜙⣙㻠㻜㻑䛾㧗ຠ⋡䛷⬺ศ໬䚹ᇵ㣴㻭㻿㻯䛾⣙㻞㻡ಸ䛾ㄪ〇ຠ⋡䚹⣙㻝㼓䛾㻌 㻌 㻌㻌 ⬡⫫⤌⧊䛛䜙䠎㐌㛫䛾ึ௦ᇵ㣴䛷㻝㻜㻤ಶ䛾㻰㻲㻭㼀䛜ᚓ䜙䜜䜛䚹㻌䝣䝷䝇䝁཯㌿㻌㻌⬡⫫⤌⧊㻌㻔㻝㻙㻞㼓㻕㻌䝁䝷䝀䝘䞊䝊㻌ฎ⌮㻌ప㏿ᗘ㻌㐲ᚰศ㞳㻌⬺ศ໬㻌ቑṪ㻌㻝㻌㐌㛫㻌䠄ኳ஭ᇵ㣴䠅㻝㻌㐌㛫㻌䠄௜╔ᇵ㣴䠅䝗䝘䞊ᖺ㱋䜔ᇶ♏⑌ᝈ䛻ᙳ㡪䛥䜜䛪ㄪ〇䛷䛝䜛㻌zᖺ㱋䠎ࠥ䠔䠍ṓ䚸㦵⢒㧼⑕䜔෌⏕୙Ⰻᛶ㈋⾑ᝈ⪅䜢ྵ䜐ᝈ⪅㻔㼚㻙㻢㻞㻕䜘䜚㻰㻲㻭㼀䛜㻌㻌 ㄪ〇䛷䛝䚸㦵䚸㌾㦵䚸⬡⫫䜈䛾ከศ໬⬟䜢♧䛩䛣䛸䜢☜ㄆ䚹㻌㻌ึ௦ᇵ㣴䛛䜙⣧ᗘ䛜㧗䛔ᆒ㉁䛺⣽⬊䛜ᚓ䜙䜜䜛㻌㻭㼐㼕㼜㼛㼟㼑㻙㼐㼑㼞㼕㼢㼑㼐㻌㼟㼠㼑㼙㻌㼏㼑㼘㼘㼟㻌㻔㻭㻿㻯㼟㻕㻌㻿㼠㼞㼛㼙㼍㼘㻌㼢㼍㼟㼏㼡㼘㼍㼞㻌㻔㻿㼂㻲㻕㻌㼒㼞㼍㼏㼠㼕㼛㼚㻌྾ᘬ⬡⫫ἲ䜢⏝䛔䛶⥅⥆ⓗ䛺㛢㙐⣔䛷䛾ㄪ〇䛜ྍ⬟㻌ኳ஭ᇵ㣴䝠䝖⬡⫫⤌⧊

Day 1

Day 2z⑭㌟⾡䛺䛹䛷⏝䛔䜙䜜䜛⬡⫫྾ᘬἲ䠄䝏䝳䝯䝉䞁䝖ἲ䠅䜢᭱㐺໬䛧䚸⣙㻝㻜㼙㼘䛾྾ᘬ⬡㻌㻌 ⫫ᾮ䛛䜙㛢㙐ᅇ㊰䛷㻰㻲㻭㼀䜢ㄪ〇䛩䜛᪉ἲ䜢㛤Ⓨ㻌㻌ᗫᲠ⬡⫫⤌⧊䜢฼⏝䛧䛯䝞䞁䜻䞁䜾䝅䝇䝔䝮䛾ᵓ⠏䛜ᐜ᫆㻌⬺ศ໬⬡⫫⣽⬊㻌㻔㻰㻲㻭㼀㻕㻌ᡂ⇍⬡⫫⣽⬊㻌zึ௦ᇵ㣴㻭㻿㻯䛿ᖹ⁥➽⣽⬊䠄㻝㻤㻚㻢㻑䠅䚸⾑⟶ෆ⓶⣽⬊㻔㻞㻚㻣㻑㻕䚸༢⌫㻔㻝㻟㻚㻟㻑㻕䛾ΰධ䛒㻌 㻌 㻌㻌 䜚䚹㻰㻲㻭㼀䛿䛣䜜䜙䛾⣽⬊䛾ΰධ⋡䛿㻜㻚㻝㻑௨ୗ䛷䛒䛳䛯䚹㻌㻌z㻴㻸㻭䝣䝹䝬䝑䝏䛧䛯௚ᐙ⛣᳜䜈䛾㐺ᛂᣑ኱䛾ྍ⬟ᛶ㻌ᇵ㣴㻰㻲㻭㼀㻌⬺ศ໬⬡⫫⣽⬊㻔㻰㻲㻭㼀㻕㻌ᡂ⇍⬡⫫⣽⬊㻌⬺ศ໬㻌⣽⬊ቑṪάᛶ㻌༢㞳䛧䛯ᡂ⇍⬡⫫⣽⬊Day 4

Day 7ኳ஭ᇵ㣴㻌௜╔ᇵ㣴㻌⬡⫫⤌⧊㻌ᖹ⁥➽⣽⬊㻌⤌⧊䝬䜽䝻䝣䜯䞊䝆㻌
௜╔ᇵ㣴㻌

㦵㻌㌾㦵㻌

⬡⫫㻌⾑⟶㻌䠄䊻䠖ᡂ⇍⬡⫫⣽⬊䛛䜙ὴ⏕䛧䛯㻰㻲㻭㼀䠅㻌ᚰ➽㻌⾑⟶ෆ⓶⣽⬊㻌≉チ㻌 ➨5055611ྕ䠄JP)䚸➨5055613ྕ(JP,GB,DE,FR)䛆㻰㻲㻭㼀䛾ㄪ〇ἲ䛸⣽⬊䛾ᙧែኚ໬䛇㻌

Matsumoto T, Kano K, et al. J Cell Physiol 215:210, 2008⬡⫫⏤᮶ᖿ⣽⬊㻔㻭㻿㻯㻕㻌ᇵ㣴㻭㻿㻯㼟㻌

㛫㉁⾑⟶ศ⏬㻔㻿㼂㻲㻕⣽⬊㻌䛆┠ⓗ䛇 DFAT䛾ከศ໬⬟䛸ྛ✀⑌ᝈ䝰䝕䝹ື≀䛻ᑐ䛩䜛἞⒪ຠᯝ䜢᳨ウ䛧䛯䚹ཎ⌮䞉᪉ἲ䞉⤖ᯝ䠄Principles, Methods, and Results䠅䠎䠊㦵෌⏕⬟䠍䠊⾑⟶෌⏕⬟䝠䝖㻌㻰㻲㻭㼀㻌

䝬䜴䝇⾑⟶ෆ⓶⣽⬊ (MS-1)

䝬䜴䝇GFP-DFAT䠏䠊ᖹ⁥➽෌⏕⬟

㻌DFAT䞉ETCP/䝁䝷䞊䝀䞁」ྜయ㻌72᫬㛫㻌 ඹᇵ㣴㻞㐌㛫㻌ASMAHoechst㦵ㄏᑟᇵᆅ㻌⓶ୗ⛣᳜㻌㦵ⱆ⣽⬊◚㦵⣽⬊

5%FBS +TGFE1
20% FBS
5% FBSᖹ⁥➽ㄏᑟᇵᆅ䛷䝠䝖DFAT䜢䠍㐌㛫ᇵ㣴䛩䜛䛸50%௨ୖ䛾⣽⬊䛜ᖹ⁥➽Į䜰䜽䝏䞁䜢Ⓨ⌧䛧䛯䚹Sakuma T et al. J Urol 182: 355, 2009䝚䞊䝗䝬䜴䝇㻌䠏㐌㛫

GFP/Hoechst

NG2/Hoechst

ETCP/䝁䝷䞊䝀䞁(Collagraft ®)GFP/NG2/Hoechst⾑⟶ෆ⓶⣽⬊䛸䛾ඹᇵ㣴䛻䜘䜚DFAT䛿50%௨ୖ䛾ຠ⋡䛷NG2㝧ᛶ䛾䝨䝸䝃䜲䝖䛻ศ໬䛧䛯䚹0W4WDFATྛ⩌㻦㻌㼚㻩㻤㻌㻌 ⏕⌮㣗ሷỈᮎᲈ⾑༢᰾⌫ᑐഃ೺ᖖ⫥䛻ᑐ䛩䜛⹫⾑⋡䠄䠂䠅㻝㻜㻜⫗║ⓗ䞉⤌⧊Ꮫⓗホ౯㻌㻌䝠䝖DFAT䜢ETCP/䝁䝷䞊䝀䞁䝇䝫䞁䝆䛻᧛✀䛧䚸䠎㐌㛫㦵ㄏᑟᇵᆅ䛷ᇵ㣴ᚋ䚸䝚䞊䝗䝬䜴䝇䛾⓶ୗ䛻⛣᳜䛧䛯䚹䠏㐌㛫ᚋ䛾⛣᳜∦䛿㦵ⱆ⣽⬊䜔◚㦵⣽⬊䜢᭷䛩䜛㢮㦵ᙧᡂ䛜ㄆ䜑䜙䜜䛯䚹୕ḟඖὶయ䝕䝞䜲䝇䛷సᡂ䛧䛯DFAT䝣䜯䜲䝞䞊䜢ᖹ⁥➽ㄏᑟᇵᆅ䛷ᇵ㣴䛩䜛䛸཰⦰⬟䜢᭷䛩䜛䝁䜲䝹≧䛾ᖹ⁥➽䛜ㄏᑟ䛥䜜䛯䚹Matsumoto T, Kano K, et al. J Cell Physiol 215:210, 2008

Hisao AY et al. PLoS One 10:e0119010, 2015

㻤㻜㻢㻜Leak point pressure (LPP)㻠㻜†DFAT㻰㻲㻭㼀㻞㻜䠆†ᮎᲈ⾑༢᰾㻼㻮㻙㻹㻺㻯⌫⏕⌮㣗ሷỈ㻯㼛㼚㼠㼞㼛㼘䠆†㻜㻜㻌㼣㼑㼑㼗㻝㻌㼣㼑㼑㼗㻞㻌㼣㼑㼑㼗㻟㻌㼣㼑㼑㼗㻠㻌㼣㼑㼑㼗†P<0.0P<0.05
P05㻌 vss ⏕⌮㣗ሷỈ⏕Ỉ,, 䠆 P<0.0P<0.05P05㻌 vss ᮎᲈ⾑༢᰾⌫ᮎ

SCID䝬䜴䝇ୗ⫥⹫⾑䝰䝕䝹䛻䝠䝖DFAT䜢➽⫗ෆᢞ୚䛩䜛䛸䚸⏕⌮㣗ሷỈ䜔ᮎᲈ⾑༢᰾⌫䛾➽⫗ෆᢞ୚䛻ẚ䜉䚸⛣᳜䠎㐌㛫ᚋ䜘䜚᭷ព䛻⾑ὶ䜢ᨵၿ䛧䛯䚹≉㢪2015-065598 ⬺ศ໬⬡⫫⣽⬊䜢᭷ຠᡂศ䛸䛩䜛⾑⟶෌⏕⒪ἲ⏝⤌ᡂ≀䜴䝃䜼⬯㦵Ḟᦆ䝰䝕䝹䛻DFAT䞉ETCP/collagen」ྜయ䜢⛣᳜䛩䜛䛸Ḟᦆ㒊䛾㦵ᐦᗘ䛾ቑຍ䛸⓶㉁㦵䛾෌⏕䛜ㄆ䜑䜙䜜䛯䚹
Kikuta S et al. Tissue Eng Part A 19:1792, 2013䝷䝑䝖⭡ᅽᛶᒀኻ⚗䝰䝕䝹䛾ᒀ㐨⤖ྜ⤌⧊䛻DFAT䜢⛣᳜䛩䜛䛸ᒀ㐨ᣓ⣙➽䛾෌⏕䛸ᒀ₃ฟᅽ(LPP)䛾ቑຍ䛜ㄆ䜑䜙䜜䛯䚹
Obinata D et al. Int J Urol 18:827, 2011䛆䜎䛸䜑䛇 DFAT䛿䚸㧗䛔⾑⟶᪂⏕⬟䚸㦵෌⏕⬟䚸ᖹ⁥➽෌⏕⬟䜢♧䛧䚸⹫⾑ᛶ⑌ᝈ䚸㦵Ḟᦆ䚸ᒀ㐨ᣓ⣙➽㞀ᐖ䛺䛹䛻ᑐ䛩䜛᭷ຠ䛺἞⒪⏝⣽⬊䛸䛺䜚䛖䜛䛣䛸䛜♧၀䛥䜜䛯䚹ᛂ⏝ศ㔝䞉⏝㏵䠄Applications䠅䛆㻰㻲㻭㼀䛻䜘䜛⣽⬊἞⒪䛜ᮇᚅ䛷䛝䜛⑌ᝈ䛇㻌㔜⑕⹫⾑ᛶ⑌ᝈ䠄ᮎᲈື⬦⑌ᝈ䛺䛹㻕㻌ⴎ⦰ᛶ⭤⬔䛻䜘䜛⭡ᅽᛶᒀኻ⚗㻌䛆㻰㻲㻭㼀⣽⬊἞⒪䛾䝫䝔䞁䝅䝱䝹䛇㻌ప䝁䝇䝖䛾⣽⬊἞⒪䞉෌⏕་⒪䛜ྍ⬟䛸䛺䜛㻌䠄኱つᶍ䛺㻯㻼㻯㻘㻌ቑṪᅉᏊ㻘㻌ᢠయ㑅ู䛺䛹ᚲせ䛺䛧䠅㻌㦵⢒㧼⑕䛻క䛖㞴἞ᛶ㦵ᢡ㻌⬨㧊ᦆയ䞉ᮎᲈ⚄⤒യᐖ䛻ᑐ䛩䜛⣽⬊἞⒪㻌㔜⑕⇕യ䛻క䛖๰യ἞⒵ಁ㐍㻌㐀⾑ᖿ⣽⬊⛣᳜䛻క䛖㻳㼂㻴㻰ண㜵䞉἞⒪㻌

䠄㦵⢒㧼⑕䜔ኚᙧᛶ㛵⠇⑕䛻క䛖㞴἞ᛶ㦵ᢡ䛺䛹䜢἞⒵䛥䛫䜛⬟ຊ䠅㻌

㌾㦵Ḟᦆ䛻ᑐ䛩䜛෌⏕་⒪㻌

㞴἞ᛶṑ࿘⑓䛻ᑐ䛩䜛ṑᵴ㦵෌⏕㻌䠄㧗㱋⪅䚸ᑠඣ䚸඲㌟≧ែ୙Ⰻ䛾ᝈ⪅䛺䛹䛻ᑐ䛩䜛⮬ᐙ䞉௚ᐙ⣽⬊἞⒪䛜ྍ⬟䠅㻌㧗㱋⪅䛾௦ㅰኚᛶ⑌ᝈ䛻ᑐᛂ䛧䚸㻽㻻㻸䜢ᨵၿ㻌୓ே䛻㐺ᛂྍ⬟䛺ᮍ᮶䛾୍⯡་⒪䛸䛺䜛ྍ⬟ᛶ㻌㻌᪥ᮏ኱Ꮫ⏘ᐁᏛ㐃ᦠ▱㈈䝉䞁䝍䞊㻌䠄㻺㼁㻮㻵㻯㻕㻌
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