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Horio F. Sakurai H. Ohsawa Y. Nakano S. Matsukura M. Fujii I.

“Functional validation and expression analysis of myotubes converted from skin fibroblasts using a simple direct reprogramming strategy.”

eNeurologicalSci. 2017;6:9-15

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`ƧDZÆ515 čJ\_¥¸½—u÷G5{z·‹ȮȥCʆʥāß_ʿDȪ qn?_MT61 ǜ̟`ƧDZƗ5ǑM> 2͌ FBS-DMEM uě@ 1 mL \M5 Ko_ 3 ǜ̟Ɨ5 10͌ FBS-DMEM 1 mL [Ő̈́ȥuÍǂM5 2 Ǖ̟Ő̈́MT6 R`Ɨ5Ő̈́ȥučĢ˚ŰŐ̈́ȥ 1 mL _ÍǂMT͍0 Ǖɜ͎ 6

ɵ͖̺ RNA ^ƷČ

RNA `ƷČačĢ˚ŰŐ̈́ȥ_řǟMTǕu 0 Ǖɜ\M5-2, 0, 3, 5, 8, 12

Ǖɜ_ʿWT6ƷČ_a NucleoSpin

RNA͍‘µžz~, ǬÎ, ǕǨ͎u

Ɇ>Z5ȨÕ`¥¹—†·_ƙ>ʿWT6

ɵ

ɵ̺͗ RT-PCR

ƷČMT RNA a PrimeScript

reagent Kit ͍ Perfect Real Time ͎ ͍‘µžz

~ , ǬÎ , ǕǨ͎uáɆMZ˿˵ĆM5 cDNA uƚT6 PCR a Takara Ex Taq

͍‘µžz~ , ǬÎ , ǕǨ͎uɆ>T6ĭƢǪÙa 94 * 2 č͍Ⱥřƣ͎`

Ɨ5 94 * 30 ɫ͍Ⱥřƣ͎ 5RrSr`xš½¶º„ȫƊ 30 ɫ͍xš½¶º

„͎ 5 72 * 1 č͍Û̝ĭƢ͎` 3 ‹•”¥u 30 ‡zƒ·5 72 * 10 č͍Û̝

ĭƢ͎ 5 4 *͍ćĨ͎[ʿWT6áɆMT¥µz«½`ƥœa˂ 2 _ɦMT6

Ą̎†º—¹½·\MZ5 hβ-actin uáɆMT6

˂

˂ 2. PCR ^{_áɆMT¥µz«½}

Gene name Number Cycle Ann. Temp. Sequence

hβ-actin 30 55 CTCTTCCAGCCTTCCTTCCT

CACCTTCACCGTTCCAGTTT

mMyoD 30 55 CTTCTATGACCCGTGTTTCGAC

CTGGGTTCCCTGTTCTGTGT

hMyoD 30 61 CACTCCGGTCCCAAATGTAG

TTCCCTGTAGCACCACACAC

hCK-M 30 55 ACATGGCCAAGGTACTGACC

TGATGGGGTCAAAGAGTTCC

hMyogenin 30 64 TAAGGTGTGTAAGGGAAGTCG CCACAGACACATCTTCCACTGT hMHC 30 61 CTGCTGAAGGAGAGGGAGCT

TGATTAGCTGGTCACACCTT

hDystrophin 30 61 GATGCACGAATGGATGACAC

TGTGCTACAGGTGGAGCTTG

hMyomaker 30 61 GAAGGAGAAGAAGGGCCTGT

CCTTCTTGTTGACCTTGGGC

ɵ͕ɺ ʊǰ

ɷčĢu˗äOqTk_5 Ad.CAG-mMyoD ƧDZƗ5Ő̈́ȥučĢ˚ŰŐ̈́

ȥ_řǟMTǕu 0 Ǖɜ\M5 -2, 0, 3, 5, 8, 12 Ǖɜ`ɷȼɎɘ̍Úşɕɂu

RT-PCR _Zɥ˙MT͍ň 8 ͎ 6«{‹̍Úş͍ m ͎uŚŅƣ̍Úş5 —̍Ú

ş͍ h ͎uĄŅƣ̍Úş`Ƽȃ\M˗äMT60 2 Ǖɜaʈǂ@x–{z·

‹¦ƒ‘½ƧDZɝĖ5 0 ǕɜačĢ˚ŰŐ̈́ȥ_řǟOqɝĖ`Ǽß`̍Ú şɕɂuǼ˕MT6 ň 8 [ˍorqn?_5 mMyoD a mMyoD ̍ÚşŰĀ Ɨ5̂mB_ɕɂC˙kor5ʈǂ@x–{z·‹¦ƒ‘½uēɆMT̍

ÚşŰĀCŁ̾^FʿtrZ>qI\uɥ˙MT6 hβ-actin aĄ̎†º—¹½

·\MZáɆMZAp5̴ƧDZ —ɚʫʓʏʵʆʥ͍ -2 Ǖɜ͎ 5

Ad.CAG-mMyoD ƧDZ —ɚʫʓʏʵʆʥčĢ˚ŰƗ 0, 3, 5, 8, 12 Ǖɜ`āZ

`ʆʥ_A>Zɕɂuɥ˙MT6ɷčĢ«½½_a¾ʳɘ_áɆKrZ>

qɷȼɎɘ̍ÚşuáɆMT [32, 33] 6 —Ą̎ɷȼɎɘčĢ˞ɺŅş[=q

hMyoD a5Ś̎BoŰĀKrT«{‹̍Úş[=q mMyoD ɕɂnpŲM̆

rZ5 3 Ǖɜ×̤_ɕɂCɥ˙KrT6hT5 MyoD ĵȁ_ɷčĢ˞ɺŅş MyoD £w¬¶½`͓ɮ[=q hMyogenin[34, 35] a 5 Ǖɜ×̤_ɕɂC˙k orT6 hMyomaker aɷʵʆʥ`ʽĴ`̬_ɕɂOqŅş[=qC [36, 37] 5 I`̍Úşl 3 Ǖɜ×̤_ɕɂOqI\Cɥ˙[DT6Iro`ʊǰnp5

ŚŅƣ mMyoD ` —ɚʫʓʏʵʆʥe`ƏĔɕɂ_np5ɷčĢ`̉ɭu

˞ɺOqˋǎ`ĄŅƣ`ŅşC˚ŰKrTI\CɦŀKrT6Ko_5 hCK-M

[38], hMHC [39], hDystrophin [1, 40, 41] l 3 Ǖɜ=q>a 5 Ǖɜ×̤npɕɂ

Cɥ˙KrZAp5ŚŅƣ MyoD ŰĀ_np5͈Ƿɷ`ȀƬŅşClToK

rT6×À`ɮ7`̍ÚşɕɂŦ͇ʊǰnp5 Ad.CAG-mMyoD _npƏĔɘ _«{‹ MyoD ̍ÚşuŰĀMT —ɚʫʓʏʵʆʥ_aĄŅƣ`ȁ7^ɷ ȼɎɘ˞ɺ̍ÚşC˚ŰKr5Rr_Ȋ>[Ȁ̃̍ÚşɕɂC˙kor5ɷ ʆʥe`Ƒ˨˵ǂuɦŀMT6

ň

ň 8. Ad.CAG-MyoD ƧDZɚʫʓʏʵʆʥ_AGqɷčĢ«½½

Ad.CAG-MyoD ƧDZɚʫʓʏʵʆʥ_AGqʉ̉Ǖǎ\ɷčĢ«½½`ɕɂuɦ

O6ɷčĢ˚ŰƗ5-2, 0, 3, 5, 8, 12 Ǖɜ` RT-PCR _nqčǮʊǰuɦO6 CK-M ͝ creatin kinase muscle isoform ^͐ MHC ͝ myosin heavy chain

%2+

%2+)'#-

%4!/&*

%

%2+$#*&*

%

%2./-+,%&*

)2+

2

ɵ

ɵ͘ɳ̱ȕĕȲ_nqĮʕʦ`Ǽ˕

ɵ͓ɺ ƈ

ɷ`ǡl̕ˌ^aToD`¾X_ɷĮʕC=q6ɷĮʕu˞ɺOqŅş_

·‰{­z~ºCɠorZAp5·‰{­z~ºaÇ_ɷűʥß_ʸ@

orZ>q6ɷűʥßaɷʓʏ`ĄóuįpŇjn?_ɕ̊MZ>q˃Ƚ`

ŃŤ[5ȅʿűɹ͍ T ɹ͎\ǁMZŠŋOq6ɷűʥßBo·‰{­z~

ºunjČOqİŪßaÇ_¶xŠºİŪß [42, 43] CɠorZAp5ɷʆʥ

`ȢĠ̱ÝC T ɹúMZɷűʥß_Útq\5ɷűʥßBo·‰{­z

~ºCnjČKr5ʆʥĄ`·‰{­z~ºȴƊC͉hq6R`·‰{­

z~ºC—¹ªšº\ʊĴOq\5xƒ“º\¬~‰º`ʊĴụŨMZ>

T—¹ª¬~‰ºCPr5xƒ“ºÀ`¬~‰ºʊĴ̎ÝC̲ČOq6RI e¬~‰ºCʊĴM5xƒ“º\¬~‰º_nqȯp˺i_npĮʕC˫I q [44] 6ɷ`̕ˌ^øD[=qĮʕʦC5 MyoD ŰĀ —ɚʫʓʏʵʆʥ[ȿ ƚ[Dq`Bu˗äOqTk5ʆʥ_̱ȕĕȲuě@5ʉ̉ˑŭMT͍ň 9 ͎ 6 ˑŭMTʆʥa Ad.CAG-mMyoD ƧDZʆʥAnc†º—¹½·ʆʥ\MZ«

{‹ɷʵʆʥ[=q C2C12 ʆʥuɆ>T6

(

( 9. +"&$'%0(

̱ȕĕȲŦ͇`ǿɍňuɦO6Ÿ·‹ɕɅˆʙ5‹z”“ĎpǠ@†º—¹½µ½5 MyoD ŰĀɚʫʓʏʵʆʥuŐ̈́MT–y”‰²ūʊMTȽƨ[̿Ɲ̜_”•

yº„M5̱ȕĕȲuě@Tǜ`ʆʥuˑŭMT6

ɵ͔ɺ Ŧ͇ǒȜ

ɵ͓̺ ʆʥŐ̈́

ɵ 5 ɳ-ɵ 2 ɺ−ɵ͓̺īȸ6

«{‹ɷʵʆʥ[=q C2C12 ʆʥ͍ATCC, Manassas, USA͎lĵȁ`ǒȜ _ZŐ̈́MT6

ɵ͔̺ ʆʥDžɮ

ʈǂ@x–{z·‹¦ƒ‘½ƧDZ`ĖǕ_5$"

"ǬÎ ǕǨ[†½•yº„MT {|· –y”‰²_ɚʫʓʏʵʆʥu (

ì{|·DžɮMT6

C2C12 ʆʥa 2.5×10

ì/{|·uÀ˖` 4 {|· C –y”‰²_DžɮM5

†º£·}º—_^qh[ 10͌ FBS-DMEM Æ[Ő̈́MT6†º£·}º—

_^WTǜȵ[5Ő̈́ȥučĢ˚ŰŐ̈́ȥ 4 mL _ÍǂMT͍1 Ǖɜ͎ 6

ɵ͕̺ ʈǂ@{z·‹¦ƒ‘½`ƧDZ

MOI 30 `ʈǂ@x–{z·‹¦ƒ‘½Ad.CAG-mMyoD-ires-Cherry A nc Ad.CAG-Cherry uRrSr —ɚʫʓʏʵʆʥ_ 2͌ FBS Ȩě DMEM 1 mL Æ[537*55͌ CO

ǪÙÁ[͓ǜ̟ƧDZKQT615 čJ\_{z·

‹ȮȥCʆʥāß_ʿDȪqn?_5¥¸½—u÷G^CoƧDZKQT6Ƨ DZƗ5ǑM> 2͌ FBS-DMEM 4 mL _Ő̈́ȥÍǂM5Ko_͕ǜ̟Ɨ5 10͌

FBS-DMEM 4 mL [ÍǂM5͔Ǖ̟Ő̈́MT6R`Ɨ5Ő̈́ȥučĢ˚ŰŐ

̈́ȥ 4 mL _ÍǂMT6̴ƧDZɚʫʓʏʵʆʥa5ʈǂ@{z·‹¦ƒ‘½ ƧDZʆʥ\ĵȁ_Ő̈́ȥ`Íǂ`iʿWT6

ɵ

ɵ͖̺ ̱ȕĕȲ

̱ȕĕȲ`ɝĖ_RrSr`Ő̈́ȥaǑM> 2 ͌ FBS-DMEM _ÍǂMT6 Ad.CAG-mMyoD-ires-Cherry ƧDZɚʫʓʏʵʆʥ5 Ad.CAG-Cherry ƧDZɚʫ ʓʏʵʆʥ5̴ƧDZɚʫʓʏʵʆʥ5 C2C12 ʆʥa5čĢ˚Ű̞ŞƗ_̱ȕ ĕȲuě@T6̱ȕĕȲa̱ȕĕȲˆʙ͍{“’̱ş , ĂɁş , ǕǨ͎uɆ

>Z 150 V, 1 Hz _Z 3 msec ĕȲ5 997 msec zº‘½ž·͍ 1 ɫ_ 1 ń`̟

̫[ĕȲ͎[ŦǓMT6

ɵ̺͗ Įʕ`˒Ǯ

ʆʥ`Įʕʦ`˗äa5ɷɹʆʥÀ`ȼť` 2 ȵ̟`Įʕˮ̯_Z˗äu ʿWT6ʆʥÀ` 2 ȵuįp5Įʕ_nq 2 ȵ̟ˮ̯u¯½‰´ºx™µz

ˆ½͍ BZ-H1M ͎ ͍ Keyence, Ŝ̢ , ǕǨ͎ _ZȬťM5ʉ̉ǜ̟\ʆʥ̟`

ˮ̯u„µ£ĢMT6

ɵ͕ɺ ʊǰ

ʆʥ`Įʕ_X>Z`˗äa5ɞ̚o`œĻMTǒȜ_ƙ>ʿWT[32, 45]6

†º—¹½·ʆʥ[=q«{‹ɷʵʆʥ` C2C12 ʆʥa5†º£·}º—_

^WTȽƨ[5Ő̈́ȥučĢ˚ŰŐ̈́ȥeřǟM5R`Ǖu 0 Ǖɜ\MT6

C2C12 ʆʥaƘ7_ɷɹuƑƬM5Ƒƨšɘ_ʅ̙Ƚ`ʆʥ\^WT6čĢ

˚Ű̞ŞBo 5 Ǖɜnp 1 ɫ̟_͓ń`̱ȕĕȲ͍150 V, on 3 msec, off 997 msec ͎ _ Ƣ N Z 5 1 ɫ ̟ _ ͓ ń ` Į ʕ C ˙ k o r T ͍ ň 10A ͎6 Ad.CAG-mMyoD-ires-Cherry ƧDZɚʫʓʏʵʆʥ[lʅ̙Ƚ`ƑƨšɘřĢ Cɥ˙Kr5 12 Ǖɜnp C2C12 \ĵȁ_̱ȕĕȲ_ƢNTĮʕC˙korT

͍ň 10B͎ 61 ɫ̟_͓ń`ĮʕCɥ˙KrT Ad.CAG-mMyoD-ires-Cherry Ƨ DZɚʫʓʏʵʆʥ[a5̱ȕĕȲ̞ŞƗ 10 ɫBo 20 ɫ_̱ȕĕȲuÆǐO q\5Rr_ĴtQZĮʕlñȋMT͍ň 10C͎ 6Ad.CAG-Cherry ƧDZɚʫʓ ʏʵʆʥAnc5̴ƧDZɚʫʓʏʵʆʥ[a̱ȕĕȲ_nqĮʕa˙kor

^BWT6Iro`ʊǰnp5MyoD ŰĀɚʫʓʏʵʆʥa̱ȕĕȲ_ĵ˞

MZĮʕMZ>qI\Cɥ˙KrT6ų5ĮʕŦ͇Æ_ʆʥ`ÝʙCśŲ`

PruɅNqTk_5őʓCIJÀCp[=WTp5IJÁCp[=WTpMZ

>q6

ň

ň 10. ̱ȕĕȲ_nqɷĮʕ

(A) C2C12 ʆʥčĢ˚Ű 5 Ǖɜ610-20 ɫa̱ȕĕȲuñȋ6

(B) Ad.CAG-mMyoD-ires-Cherry ƧDZɚʫʓʏʵʆʥčĢ˚Ű 14 Ǖɜ6

(C) Ad.CAG-mMyoD-ires-Cherry ƧDZɚʫʓʏʵʆʥčĢ˚Ű 14 Ǖɜ610-20 ɫa̱

ȕĕȲuñȋ6

&./*!#5)

&)#.

&./*!#5)

&)#.

&./*!#5)

&)#.

ɵ

ɵ͙ɳ ·‰{­z~ºĠƨ`ɥ˙

ɵ͓ɺ ƈ

MyoD ŰĀ —ɚʫʓʏʵʆʥ[aɷűʥßCƑƬKrZ>q`B˗äO qTk_5·‰{­z®½Šº„ȜuɆ>ZǼ˕uʿWT6·‰{­z

®½Šº„Ȝ\a5·‰{­z~º\ʊĴOq\ʼýuɕOq¥¹½¤͍

·‰{­ʼý¥¹½¤͎uáɆMZ5ʆʥĄ[`·‰{­z~º`Ġƨu ɋù\MZˑȬıʦ\OqǒȜ[=q6·‰{­¥¹½¤aʆʥĄ`·

‰{­z~ºȴƊ_ƢNZʼýƏƊCřtqTk5R`ʼýƏƊ`ƊĴ>_

np·‰{­z~º`ȴƊuˑŭOqI\C[Dq6ÑńáɆMT·‰

{­ʼý¥¹½¤ Fluo 4-AM aȸůý`Ğ˫ȝ̝ 495 nm _A>Z5 518 nm Õ˻[ʼýCˑŭKrq6 Fluo 4-AM ` AM ax—‰®“·ő͍ AM ő͎

u˂M5·‰{­z~ºu¸½—Oq̎č͍·©‰·ő͎Cx—

®“·ő[éˣKrZ>q6Ir_npʆʥ̀̉ƣuƻTQZ>q6Ko_5

¥¹½¤aʆʥĄ_Āq\5 AM őCʆʥĄ}‹•µ½Ž_nWZěȖč˒

uİG5·‰{­z~ºu¸½—[DqȀ̃\^p5BXʆʥŚeȰr ČM_FF^q\>?ȼƞuƻX͍ :aNkZ`ʆʥĄ Ca

Ȭť¥¹—†·;

ĵÐĢšɣɯư , ȹǨ , ǕǨ͎6ʆʥĄ[`·‰{­z~º`njČuˑŭO qTk_5ɷűʥßÀ`İŪßuĕȲOqʹė5O^tV ATP Ancƒ¹¹

£|½·˚Űß͍ň 11 ͎_ŮOqĭƢu˗äMT6 ʆʥŚ ATP a G ʻɗ

˨ăƔō ATP İŪß_ʊĴM5¨‹¨¶Ÿ½Ž C uȢƣĢKQq6R`ʊǰ

¨‹£w“Š·z‰—½·Ê¶º̓CŠx‰·„¶¹½·\z‰—½

·¿¶º͍̓IP

͎_č˒Kr5ʆʥĄ[ IP

ȴƊCÀǗOq6ȴƊCÀǗM T IP

CɷűʥßÀ` IP

İŪßuĕȲM5ɷűʥߥ_ʸ@orZ>q·

‰{­z~ºCʆʥ˨Ą_njČKrq[46,47]64-ƒ¹¹-3-}“·£|½·

͍4-CEP͎Anc 4-ƒ¹¹-m-ƒ¸½·͍4-CmC͎aɷűʥßÀ`¶xŠ

ºİŪßuĕȲM5·‰{­z~ºunjČKQq͍ň 12͎[48]6Iro`

˘ʹu Ad.CAG-mMyoD-ires-Cherry ƧDZƗ 16 Ǖɜ`ɚʫʓʏʵʆʥ\̴ƧDZ ɚʫʓʏʵʆʥeȨěM5Ȕ˷Ǽ˕MT6

ň 11. ɷűʥßÀ`İŪßĕȲʹ

ň

ň 12. ^{ɷűʥßÀ`İŪß}

ɷűʥßÀ`İŪß_X>ZɦO6áɆMTʹė ATP54-CEP54-CmC aɷűʥß À`İŪßuĕȲOq6ATP a IP

İŪßu̟ǁɘ_ĕȲOqI\[5 4-CEP Anc

4-CmC a¶xŠºİŪßuĕȲOqI\[ɷűʥßnp·‰{­z~ºunjČ

KQq6

ɵ͔ɺ Ŧ͇ǒȜ

ɵ͓̺ ʆʥŐ̈́Ancʈǂ@{z·‹¦ƒ‘½`ƧDZ

ɵ 5 ɳ - ɵ 2 ɺ - ɵ 1 ̺Ancɵ 2 ̺īȸ6

ɵ

ɵ͔̺ Ȩěʹė`˞ˊ

1 500 μ M 4-chloro-3-ethylphenol ͍ 4-CEP ͎Ȯȥ

0.783 g ` 4-CEP ͍ Sigma-Aldrich, ǬÎ , ǕǨ͎u 200 μL ` DMSO ͍™

µz•‹ƒ , Î̏ , ǕǨ͎[Ȯ˒M5Ȯ˒ȥu Recording Medium ͍ Fluo 4-AM special packaging

kit ͍ĵÐĢš , ȹǨ , ǕǨ͎ ͎_ȧĴM5ā̗u 10 mL \M T6

2 500 μ M 4-chloro-m-cresol͍4-CmC͎Ȯȥ

0.71 g ` 4-CmC ͍Sigma-Aldrich, ǬÎ, ǕǨ͎ u 200 μ L ` DMSO [Ȯ˒M5 Ȯ˒ȥu Recording Medium͍Fluo 4-AM special packaging

kit͎_ȧĴM5ā

̗u 10 mL \MT6

3 100 μ M ATP Ȯȥ

ATP͍MP Biomedicals, ǬÎ, ǕǨ͎ 60.5 mg u Recording Medium [Ȯ˒

Mā̗u 10 mL \M510 mM ATP Ȯȥu˞ˊMT610 mM ATP Ȯȥ 50 μ L u Recording Medium ͍Fluo 4-AM special packaging

kit͎\ȧĴM5ā̗ 5 mL

\MZ5100 μ M ATP Ȯȥ\MT6

ɵ͕̺ ·‰{­z~º`Ȭť

·‰{­z®½Šº„a Fluo4-AM special packaging

kit uɆ>Z5ȨÕ

`¥¹—†·_ƙWZʿWT6ʆʥĄ·‰{­z~ºȴƊuȬťOqTk

_5 Fluo4-AM uȨěM 37 *[ 1 ǜ̟̳ʙMZ5ʆʥĄ_įp˺hQT6R

`Ɨ5ʆʥuȡ>5ÕŶ` Recording Medium _ÍǂM5ˑŭuʿWT6ˑ ŭ̞ŞƗ 10 ɫƗ_RrSr`ʹėuȨěMT6áɆMTʹėa 500 μ M 4-CEP, 500 μM 4-CmC, 100 μ M ATP [=q6ʼýƏƊa BZ-X Analyzer

͍Keyence, Ŝ̢, ǕǨ͎ _Z˔ȬMT6ĵ¾–y”‰²np 15 ì`ʆʥu

̌ƶM5ʼýƏƊ`ǡŜðuǎðĢMT6ij„·½¥`ʼýƏƊ`Žu

Student’s t-test _ZȔ˷MT6 P < 0.05 ,# */

ɵ͕ɺ ʊǰ

Ad.CAG-mMyoD-ires-Cherry ƧDZ —ɚʫʓʏʵʆʥ\̴ƧDZɚʫʓʏʵ ʆʥ_AGq5ƒ¹¹£|½·˚ŰßAnc ATP Ȩěǜ`ʆʥĄ·‰{

­z~ºȴƊ`řĢu˗äMT6 100 μ M ATP 5 500 μ M 4-CEP 5 500 μ M 4-CmC Ȩ ěǜ`ʊǰuň 13 _ɦO6Ćɞaij˘ʹȨěƗ`ʼýƏƊǡűǜ͍˘ʹȨě Ė͎AncʼýƏƊǡŜǜuɦMZ>q͍ň 13A ͎ 6ʆʥŚ ATP a IP

İŪß

`̟ǁɘĕȲė[=qC5ʹėȨěƗ5 MyoD ̴ŰĀɚʫʓʏʵʆʥ[aʼ ýƏƊ`ŖŜaˑȬ[D^BWT`_ŮM5 MyoD ŰĀɚʫʓʏʵʆʥ[a ʼýƏƊ`ŖŜCɥ˙KrT6¶xŠºİŪßuĕȲOq 4-CEP Anc

4-CmC ȨěƗ5 MyoD ̴ŰĀɚʫʓʏʵʆʥ_Ȕf5 MyoD ŰĀɚʫʓʏʵ

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ň 13. ·‰{­z®½Šº„

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(C) ǡŜʼýƏƊ`Ȕ˷6 N=15 6 *P < 0.05 6

MyoD͍+͎: MyoD ŰĀ —ɚʫʓʏʵʆʥ, MyoD͍-͎: ̴ŰĀ —ɚʫʓʏʵʆʥ

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(B) Ad.CAG-mMyoD-ires-EGFP Ƨ DZ ɚ ʫ ʓ ʏ ʵ ʆ ʥ 5 ̖ Ʌ ō « { ‹ ɷ ʵ ʆ ʥ 5 Ad.CAG-EGFP ƧDZɚʫʓʏʵʆʥ`ƂŌ̂Ɗ6ða±SE6 n=156 *P < 0.05 **P < 0.05

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