Clumps of Mesenchymal Stem Cell/Extracellular Matrix Complexes Generated with Xeno-Free Conditions Facilitate Bone Regeneration via Direct and Indirect Osteogenesis.

概要

Ꮫ ఩ ㄽ ᩥ

Clumps of Mesenchymal Stem Cell/Extracellular
Matrix Complexes Generated with Xeno-Free
Conditions Facilitate Bone Regeneration via Direct
and Indirect Osteogenesis.
(ࢮࣀࣇ࣮ࣜ᮲௳ୗ࡟స〇ࡋࡓ㛫ⴥ⣔ᖿ⣽⬊㞟ሢ Clumps of mesenchymal
stem cell/extracellular matrix complexes ࡣ⮬㌟ࡢ㦵ศ໬࡜࣍ࢫࢺ⣽⬊ࡢ
㦵ᙧᡂㄏᑟࢆ௓ࡋ࡚㦵⤌⧊෌⏕ࢆಁ㐍ࡍࡿ)

ᮏụ ⥲ኴ
ᗈᓥ኱Ꮫ኱Ꮫ㝔་ṑ⸆ಖ೺Ꮫ◊✲⛉
༤ኈㄢ⛬ ་ṑ⸆Ꮫᑓᨷ
㸰㸮㸯㸷ᖺᗘ

୺ᣦᑟᩍဨ㸸ᰩཎ ⱥぢ ᩍᤵ
㸦་⣔⛉Ꮫ◊✲⛉ ṑ࿘⑓ែᏛ㸧
1

ᮏㄽᩥࡢせ᪨ࡣ௨ୗࡢᏛ఍࡟࠾࠸࡚Ⓨ⾲ࡋࡓࠋ

➨ 18 ᅇ ᪥ᮏ෌⏕་⒪Ꮫ఍⥲఍
㸦2019 ᖺ 3 ᭶ රᗜ㸧
➨ 62 ᅇ ᫓Ꮨ᪥ᮏṑ࿘⑓Ꮫ఍Ꮫ⾡኱఍
㸦2019 ᖺ 5 ᭶ ⚄ዉᕝ㸧

2

ㅰ ㎡

ᮏ◊✲࡟㝿ࡋࠊᚚᣦᑟ࡞ࡽࡧ࡟ᚚ㧗㜀ࢆ㈷ࡾࡲࡋࡓᮏᏛ኱Ꮫ㝔 ་⣔⛉Ꮫ◊✲⛉ ṑᏛᑓ
㛛ࣉࣟࢢ࣒ࣛ ṑ࿘⑓ែᏛ◊✲ᐊ ᰩཎ ⱥぢ ᩍᤵ࡟῝⏒࡞ࡿឤㅰࡢពࢆ⾲ࡋࡲࡍࠋࡲࡓࠊ◊
✲㐙⾜࡞ࡽࡧ࡟ᮏㄽᩥసᡂ࡟࠾࠸࡚ࠊᚚᩍ♧ࠊᚚ㠴᧡㡬ࡁࡲࡋࡓᮏᏛ኱Ꮫ㝔 ་⣔⛉Ꮫ◊✲
⛉ ṑᏛᑓ㛛ࣉࣟࢢ࣒ࣛ ṑ㧊⏕≀Ꮫ◊✲ᐊ ᰘ ⚽ᶞ ᩍᤵࠊṑ⛉▹ṇᏛ◊✲ᐊ ㇂ᮏ ᖾኴ㑻
ᩍᤵࠊ⏕యศᏊᶵ⬟Ꮫ◊✲ᐊ ᐟ༡ ▱బ ᩍᤵࠊ⏕యᮦᩱᏛ◊✲ᐊ ຍ⸨ ຌ୍ ᩍᤵࠊᗈᓥ኱
Ꮫ⑓㝔 ཱྀ⭍⥲ྜデ⒪⛉ Ἑཱྀ ᾈஅ ᩍᤵ࡟῝ཌ࡞ࡿㅰពࢆ⾲ࡋࡲࡍࠋࡉࡽ࡟ᮏ◊✲ࡢ㐙⾜ࠊ
࠾ࡼࡧᮏㄽᩥࡢసᡂ࡟࠶ࡓࡾ⤊ጞᚚᣦᑟࠊᚚຓゝࢆ㡬ࡁࡲࡋࡓᮏᏛ኱Ꮫ㝔 ་⣔⛉Ꮫ◊✲⛉
ṑᏛᑓ㛛ࣉࣟࢢ࣒ࣛ ṑ࿘⑓ែᏛ◊✲ᐊ ຍ἞ᒇ ᖿே ຓᩍ࡟῝ࡃឤㅰ࠸ࡓࡋࡲࡍࠋ
ࡲࡓࠊᮏ◊✲ࢆ㐍ࡵࡿ࡟㝿ࡋከ኱࡞ࡿᚚᨭᣢࢆ㡬ࡁࡲࡋࡓᮏᏛ኱Ꮫ㝔 ་⣔⛉Ꮫ◊✲⛉ࡢ
ⓙᵝ࡟ཌࡃᚚ♩⏦ࡋୖࡆࡲࡍࠋ
᭱ᚋ࡟ࠊຮᏛࠊ◊✲ࡢᶵ఍ࢆ୚࠼ࡿ࡜ඹ࡟ࠊᖖ࡟⚾ࢆᨭ࠼࡚ࡃࢀࡓ∗࣭㑥᫂ࠊẕ࣭⿱Ꮚ࡟
ᚰ࠿ࡽឤㅰ⮴ࡋࡲࡍࠋ

2020 ᖺ 1 ᭶
ᗈᓥ኱Ꮫ኱Ꮫ㝔 ་ṑ⸆ಖ೺Ꮫ◊✲⛉
་ṑ⸆Ꮫᑓᨷ ṑ࿘⑓ែᏛ◊✲ᐊ
ᮏụ ⥲ኴ

3

┠ ḟ

➨ 1 ❶ ⥴ ㄽ ······························································································ 7

➨ 2 ❶ Xeno-free/Serum-free ᮲௳ୗ࡛స〇ࡋࡓ C-MSCs ࡢᛶ㉁ ························ 11
  ➨ 1 ⠇ Xeno-free/Serum-free ᮲௳ୗ࡛ࡢ C-MSCs స〇 ······························· 11
➨ 1 㡯 ᴫ せ ···················································································· 11
➨ 2 㡯 ᮦᩱ࡜᪉ἲ ············································································ 11
1. ࣄࢺ MSCs ࡢᇵ㣴ἲ··································································· 11
2. Xeno-free/Serum-free ᮲௳ୗ࡛ࡢ⣽⬊㞟ሢ C-MSCs ࡢస〇ἲ·············· 11
3. ࣇ࣮ࣟࢧ࢖ࢺ࣓ࢺ࣮ࣜ (FCM) ゎᯒ ················································ 12
4. ⤌⧊Ꮫⓗゎᯒ ·············································································· 12
1) ࣐࣊ࢺ࢟ࢩ࣭࢚ࣜࣥ࢜ࢪࣥ (HE) ᰁⰍ ········································ 12
2) ⺯ගච␿ᰁⰍ ·········································································· 13
5. ࢔࣏ࢺ࣮ࢩࢫ⣽⬊ࡢ᳨ฟ ······························································· 14
➨ 3 㡯 ⤖ ᯝ ···················································································· 14
  ➨ 2 ⠇ Xeno-free/Serum-free ᮲௳ୗ࡛స〇ࡋࡓ C-MSCs ࡢ㦵⤌⧊෌⏕⬟ࡢ᳨ウ
······························································································ 15
➨ 1 㡯 ᴫ せ ···················································································· 15
➨ 2 㡯 ᮦᩱ࡜᪉ἲ ············································································ 15
1. SCID ࣐࢘ࢫ㢌⵹ෙ 1.6mm 㦵Ḟᦆࣔࢹࣝ࡬ࡢ C-MSCs ⛣᳜ ················ 15
2. micro CT ゎᯒἲ ·········································································· 15
3. HE ᰁⰍ ······················································································ 16
4. ⤫ィゎᯒ ···················································································· 16
4

➨ 3 㡯 ⤖ ᯝ ···················································································· 16
➨ 3 ⠇ ᑠ ᣓ ························································································ 17

➨ 3 ❶ C-MSCs ࡢ㦵⤌⧊෌⏕ᶵᗎࡢゎᯒ ························································ 18
➨ 1 ⠇ C-MSCs ⛣᳜࡟ࡼࡿ㦵⤌⧊෌⏕㐣⛬࡟࠾ࡅࡿࢻࢼ࣮⣽⬊࡜࣍ࢫࢺ⣽⬊
ࡢศᕸゎᯒ ················································································· 18
➨ 1 㡯 ᴫ せ ···················································································· 18
➨ 2 㡯 ᮦᩱ࡜᪉ἲ ············································································ 18
1. ࣄࢺ vimentin ࡟ᑐࡍࡿ⺯ගච␿ᰁⰍἲ ········································· 18
2. ⤫ィゎᯒ ··················································································· 19
➨ 3 㡯 ⤖ ᯝ ···················································································· 19
➨ 2 ⠇ ᪂⏕㦵ᵝ⤌⧊ࡢ㦵㉁ࡢゎᯒ···························································· 19
➨ 1 㡯 ᴫ せ ···················································································· 19
➨ 2 㡯 ᮦᩱ࡜᪉ἲ ············································································ 19
1. Azan ᰁⰍἲ ················································································ 19
2. ⺯ගච␿ᰁⰍἲ ··········································································· 20
1) ࣄࢺ㦵ᇶ㉁㛵㐃ࢱࣥࣃࢡࡢච␿ᰁⰍἲ ·········································· 20
2) 㦵⣽⬊ࡢච␿ᰁⰍἲ ·································································· 21
➨ 3 㡯 ⤖ ᯝ ···················································································· 22
➨ 3 ⠇ ⬺⣽⬊ฎ⌮ࢆ⾜ࡗࡓ C-MSCs (Decell-C-MSCs) ࡢ㦵⤌⧊෌⏕⬟ࡢ᳨ウ
······························································································· 23
➨ 1 㡯 ᴫ せ ···················································································· 23
➨ 2 㡯 ᮦᩱ࡜᪉ἲ ············································································ 23
1. C-MSCs ࡢ⬺⣽⬊ฎ⌮ἲ ································································ 23
5

2. Decell-C-MSCs ࡢᛶ㉁ホ౯ ···························································· 23
1) HE ᰁⰍ ·················································································· 23
2) ⺯ගච␿ᰁⰍ ··········································································· 23
3) ኚᛶࢥ࣮ࣛࢤࣥᰁⰍ ·································································· 24
3. NOD/SCID ࣐࢘ࢫ㢌⵹ෙ 1.6mm 㦵Ḟᦆࣔࢹࣝ࡬ࡢ Decell-C-MSCs ⛣᳜
································································································ 24
4. micro CT ゎᯒἲ ·········································································· 24
5. ⤌⧊Ꮫⓗほᐹ ·············································································· 24
1) HE ᰁⰍ ··················································································· 24
2) Azan ᰁⰍ ················································································ 24
3) ⺯ගච␿ᰁⰍ ··········································································· 24
6. ...

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