iPS細胞・ES細胞・幹細胞などへのエレクトロポレーションによる遺伝子導入

NEPA21

ELEPO21

NEPAｷｭﾍﾞｯﾄ電極

ｷｭﾍﾞｯﾄ電極用ﾁｬﾝﾊﾞｰ･ﾗｯｸ

ﾓﾆﾀﾘﾝｸﾞｼｽﾃﾑ

iPS細胞を始め様々な幹細胞にも、高価な専用試薬・バッファー不要で、簡単かつ非常に効率の良いトランスフェクションが可能です。

iPS細胞・ES細胞・幹細胞などへのエレクトロポレーションによる遺伝子導入

アプリケーション

iPS細胞・ES細胞・幹細胞への遺伝子導入例

ヒトiPS細胞

エレクトロポレーション後3日目

ヒトiPS細胞

エレクトロポレーション後2日目　（コロニーの部分に導入されている 。）

エレクトロポレーション後7日目　（継代後も、iPS細胞コロニーでGFPが発現している 。）

マウスES細胞

生存率：74％

導入効率：88％

マウスNeurospheres

生存率：90%

導入効率：75%

ヒトiPS細胞　胚葉体（接着状態）

 

 

iPS細胞の作製

疾患iPS細胞（LQT）の作製

B細胞へのエピソーマルベクターを用いた初期化因子の導入

疾患iPS細胞（LQT）：EP後およそ1ヶ月のiPS細胞のコロニー。（対物 x5）

アルカリホスファターゼ染色。（左：対物 x4　右：対物 x10）

疾患（LQT）iPSコロニーの免疫染色。
未分化マーカー TRA1-60 (細胞表面抗原), OCT4 （核）の発現は非常に良好である。
青:DAPI（核）　緑（左）:TRA1-60 　緑（右）:OCT4　赤:ファロイジン（重合型アクチン）

東京女子医科大学　国際分子細胞免疫研究センター　循環器小児科　中西敏雄研究室　ご提供

 

NEPA21トランスフェクションによるヒトiPS細胞のCRISPR活性評価

NEPA21 iPS細胞　プロトコル

  1. ヒトiPS細胞株におけるNEPA21スーパーエレクトロポレーター（ネッパジーン社）を用いたトランスフェクション条件の最適化。Poring Pulseの電圧とパルス幅を検討した。　EGFP発現プラスミドをトランスフェクションし、LSRFortessaセルアナライザー（BD社）を用いて陽性細胞の割合を解析した。
  2. トランスフェクション手順の概略
  3. Cas9発現ベクターpHL-EF1a-SphcCas9-iP-A（Addgene ID: 60599）とsgRNA発現ベクターpHL-H1-ccdB-mEF1a-RiH（Addgene ID: 60601）をiPS細胞にエレクトロポレーションした後、内在性ジストロフィン遺伝子の切断活性をT7EIアッセイで評価した。切断されたバンドの強度（◀赤矢印）をTapeStation（Agilent）で検出し、その値はゲルエレクトログラム画像の下に示す。
  4. Cと同じゲノムDNAサンプルを制限酵素XcmI（50-CCANNNNN＾NNTGG-30）を用いたRFLPで分析した。切断されていないバンドの強度は、◀赤矢印に示す。gRNA5は、XcmI部位から遠く離れた場所で切断するため、RFLPアッセイでは切断活性は検出されなかった。

京都大学　iPS細胞研究所　臨床応用研究部門　李紅梅先生・堀田秋津先生　提供

 

iPS細胞・ES細胞・幹細胞への遺伝子導入結果一覧

細胞名をクリック頂きますと、細胞の写真がご覧頂けます。

細胞名生存率

導入
効率

 細胞名生存率

導入
効率

ヒトiPS細胞 (201B7)86%70% ヒトiPS細胞 (201B7) 94%80%
ヒトiPS細胞   ヒトiPS細胞  
ヒトiPS細胞 (201B7)   ヒトiPS細胞 (201B7)  
ヒトiPS細胞69%80% ヒトiPS細胞 73%
ヒトiPS細胞由来神経細胞93%54%    
ヒトES細胞（H9 p.51）55%55% ヒトES細胞  
ヒト間葉系幹細胞（プライマリー）78%75%  ヒト間葉系幹細胞（hMSC）70%80%
ヒト神経幹細胞97%95% ヒト神経幹細胞80%83%
ヒト乳歯歯髄幹細胞（SHED）90%92% ヒト造血幹細胞を含む有核細胞
（精製前段階）		73%90%
マウスiPS細胞70%50% マウスES細胞80%75%
マウス ES細胞80%68% マウス ES細胞 74%88%
マウス ES細胞 (129系統 R1/E)80%90%  マウスES細胞70%100%
マウス ES細胞80%90% マウスiPS細胞由来神経幹細胞 86%
マウス神経幹細胞90%80% マウス神経幹細胞（プライマリー）80%60%
マウス神経幹細胞塊（Neurospheres）90%75% マウス神経幹細胞塊（Neurospheres）  
TS マウス栄養膜幹細胞59%47% C3H/10T1/2 マウス間葉系幹細胞70%85%
マウス間葉系幹細胞99％89％ マウス造血幹細胞（c-Kit陽性細胞）66％45％
ラットES様細胞70%76%  ラットWDA ES様細胞60%80％

文献

  • ヒトiPS細胞

CRISPR loss of function screening to identify genes involved in human primordial germ cell-like cell development

Hwang YS, Seita Y, Blanco MA, Sasaki K.

PLoS Genet. 2023 Dec 13;19(12):e1011080.

  • ヒトiPS細胞

Uniform transgene activation in Tet-On systemsdepends on sustained rtTA expression

Otomo J, Woltjen K, Sakurai H.

iScience. 2023 Aug 19;26(10):107685.

  • ヒトiPS細胞

Core-shell hydrogel microfiber-expanded pluripotent stem cell-derived lung progenitors applicable to lung reconstruction in vivo

Ikeo S, Yamamoto Y, Ikeda K, Sone N, Korogi Y, Tomiyama L, Matsumoto H, Hirai T, Hagiwara M, Gotoh S.

Biomaterials. 2021 Sep:276:121031.

  • ヒトiPS細胞

A muscle fatigue-like contractile decline was recapitulated using skeletal myotubes from Duchenne muscular dystrophy patient-derived iPSCs

Uchimura T, Asano T, Nakata T, Hotta A, Sakurai H.

Cell Rep Med. 2021 Jun 4;2(6):100298.

  • ヒトiPS細胞

Successful Correction of ALD Patient-derived iPSCs Using CRISPR/Cas9

Eul Sik Jung, Zhejiu Quan, Mi-Yoon Chang, Wonjun Hong, Ji Hun Kim, Seung Hyun Kim, Seungkwon You, Dae-Sung Kim, Jiho Jang, Sang-Hun Lee,Hyongbum (Henry) Kim, Hoon Chul Kang

bioRxiv February 25, 2020

  • ヒトiPS細胞

In vitro characterization of the human segmentation clock

Margarete Diaz-Cuadros, Daniel E Wagner, Christoph Budjan, Alexis Hubaud, Oscar A Tarazona, Sophia Donelly, Arthur Michaut, Ziad Al Tanoury, Kumiko Yoshioka-Kobayashi, Yusuke Niino, Ryoichiro Kageyama, Atsushi Miyawaki, Jonathan Touboul, Olivier Pourquié

Nature. 2020 Apr;580(7801):113-118.

  • ヒトiPS細胞
  • リプログラミングヒトiPS細胞

Establishment of a Non-Integrate iPS Cell Line CSUASOi002-A, From Urine-Derived Cells of a Female Patient With Macular Corneal Dystrophy Carrying Compound Heterozygous CHST6 Mutations

Jing Y, Zhou Y, Wang C, Liu J, Guo Y, Mao S, Chan HF, Tang S, Chen J.

Stem Cell Res. 2019 Dec:41:101598.

  • ヒトiPS細胞

Functional Evaluation of the Pathological Significance of MEFV Variants Using Induced Pluripotent Stem Cell-Derived Macrophages

Shiba T, Tanaka T, Ida H, Watanabe M, Nakaseko H, Osawa M, Shibata H, Izawa K, Yasumi T, Kawasaki Y, Saito MK, Takita J, Heike T, Nishikomori R.

J Allergy Clin Immunol. 2019 Nov;144(5):1438-1441.e12.

  • ヒトiPS細胞

Persistent Epigenetic Memory Impedes Rescue of the Telomeric Phenotype in Human ICF iPSCs Following DNMT3B Correction

Toubiana S, Gagliardi M, Papa M, Manco R, Tzukerman M, Matarazzo MR, Selig S.

Elife. 2019 Nov 20:8:e47859.

  • ヒトiPS細胞

Establishment of SLC15A1/PEPT1-Knockout Human-Induced Pluripotent Stem Cell Line for Intestinal Drug Absorption Studies

Kawai K, Negoro R, Ichikawa M, Yamashita T, Deguchi S, Harada K, Hirata K, Takayama K, Mizuguchi H.

Mol Ther Methods Clin Dev. 2019 Nov 21:17:49-57

  • マウスES細胞

Developmental Analyses of Mouse Embryos and Adults Using a Non-Overlapping Tracing System for All Three Germ Layers

Serizawa T, Isotani A, Matsumura T, Nakanishi K, Nonaka S, Shibata S, Ikawa M, Okano H.

Development. 2019 Nov 4;146(21):dev174938.

  • マウスES細胞

Preparation and electroporation of Cas12a/Cpf1-guide RNA complexes for introducing large gene deletions in mouse embryonic stem cells

Kissling L, Monfort A, Swarts DC, Wutz A, Jinek M

Methods Enzymol. 2019;616:241-263.

  • ヒトiPS細胞

Genome-wide microhomologies enable precise template-free editing of biologically relevant deletion mutations

Grajcarek J, Monlong J, Nishinaka-Arai Y, Nakamura M, Nagai M, Matsuo S, Lougheed D, Sakurai H, Saito MK, Bourque G, Woltjen K.

Nat Commun. 2019 Oct 24;10(1):4856.

  • ヒトiPS細胞

A Versatile Toolbox for Knock-In Gene Targeting Based on the Multisite Gateway Technology

Yoshimatsu S, Sone T, Nakajima M, Sato T, Okochi R, Ishikawa M, Nakamura M, Sasaki E, Shiozawa S, Okano H.

PLoS One. 2019 Aug 27;14(8):e0221164.

  • ヒトiPS細胞

Core Transcription Factors Promote Induction of PAX3-Positive Skeletal Muscle Stem Cells

Sato T, Higashioka K, Sakurai H, Yamamoto T, Goshima N, Ueno M, Sotozono C.

Stem Cell Reports. 2019 Aug 13;13(2):352-365.

  • ヒトiPS細胞

Phenotype-Based High-Throughput Classification of Long QT Syndrome Subtypes Using Human Induced Pluripotent Stem Cells

Yoshinaga D, Baba S, Makiyama T, Shibata H, Hirata T, Akagi K, Matsuda K, Kohjitani H, Wuriyanghai Y, Umeda K, Yamamoto Y, Conklin BR, Horie M, Takita J, Heike T.

Stem Cell Reports. 2019 Aug 13;13(2):394-404.

  • ヒトiPS細胞

Aberrant Axon Branching via Fos-B Dysregulation in FUS-ALS Motor Neurons

Akiyama T, Suzuki N, Ishikawa M, Fujimori K, Sone T, Kawada J, Funayama R, Fujishima F, Mitsuzawa S, Ikeda K, Ono H, Shijo T, Osana S, Shirota M, Nakagawa T, Kitajima Y, Nishiyama A, Izumi R, Morimoto S, Okada Y, Kamei T, Nishida M, Nogami M, Kaneda S, Ikeuchi Y, Mitsuhashi H, Nakayama K, Fujii T, Warita H, Okano H, Aoki M.

EBioMedicine. 2019 Jul:45:362-378.

  • ヒト歯髄幹細胞(DPSCs)

Novel Gain-Of-Function Mutation of TRPV4 Associated With Accelerated Chondrogenic Differentiation of Dental Pulp Stem Cells Derived From a Patient With Metatropic Dysplasia

Nonaka K, Han X, Kato H, Sato H, Yamaza H, Hirofuji Y, Masuda K.

Biochem Biophys Rep. 2019 May 17:19:100648.

  • ヒトiPS細胞

Function and Immunogenicity of Gene-corrected iPSC-derived Hepatocyte-Like Cells in Restoring Low Density Lipoprotein Uptake in Homozygous Familial Hypercholesterolemia

Okada H, Nakanishi C, Yoshida S, Shimojima M, Yokawa J, Mori M, Tada H, Yoshimuta T, Hayashi K, Yamano T, Hanayama R, Yamagishi M, Kawashiri MA

Sci Rep. 2019 Mar 18;9(1):4695.

  • ヒトiPS細胞

Verification and rectification of cell type-specific splicing of a Seckel syndrome-associated ATR mutation using iPS cell model.

Ichisima J, Suzuki NM, Samata B, Awaya T, Takahashi J, Hagiwara M, Nakahata T, Saito MK.

J Hum Genet. 2019 May;64(5):445-458.

  • ヒトES細胞

The Lineage-Specific Transcription Factor CDX2 Navigates Dynamic Chromatin to Control Distinct Stages of Intestine Development

Kumar N, Tsai YH, Chen L, Zhou A, Banerjee KK, Saxena M, Huang S, Toke NH, Xing J, Shivdasani RA, Spence JR, Verzi MP.

Development. 2019 Mar 1;146(5):dev172189.

  • ヒトiPS細胞

MiR-33a is a therapeutic target in SPG4-related hereditary spastic paraplegia human neurons

Nakazeki F, Tsuge I, Horie T, Imamura K, Tsukita K, Hotta A, Baba O, Kuwabara Y, Nishino T, Nakao T, Nishiga M, Nishi H, Nakashima Y, Ide Y, Koyama S, Kimura M, Tsuji S, Naitoh M, Suzuki S, Izumi Y, Kawarai T, Kaji R, Kimura T, Inoue H, Ono K

Clin Sci (Lond). 2019 Feb 22;133(4):583-595.

  • ヒトiPS細胞

Increased Cytotoxicity of Herpes Simplex Virus Thymidine Kinase Expression in Human Induced Pluripotent Stem Cells

Iwasawa C, Tamura R, Sugiura Y, Suzuki S, Kuzumaki N, Narita M, Suematsu M, Nakamura M, Yoshida K, Toda M, Okano H, Miyoshi H.

Int J Mol Sci. 2019 Feb 14;20(4):810.

  • ヒトiPS細胞

A Patient-Derived iPSC Model Revealed Oxidative Stress Increases Facioscapulohumeral Muscular Dystrophy-Causative DUX4

Sasaki-Honda M, Jonouchi T, Arai M, Hotta A, Mitsuhashi S, Nishino I, Matsuda R, Sakurai H.

Hum Mol Genet. 2018 Dec 1;27(23):4024-4035.

  • ヒトiPS細胞

Enhancing T Cell Receptor Stability in Rejuvenated iPSC-Derived T Cells Improves Their Use in Cancer Immunotherapy

Minagawa A, Yoshikawa T, Yasukawa M, Hotta A, Kunitomo M, Iriguchi S, Takiguchi M, Kassai Y, Imai E, Yasui Y, Kawai Y, Zhang R, Uemura Y, Miyoshi H, Nakanishi M, Watanabe A, Hayashi A, Kawana K, Fujii T, Nakatsura T, Kaneko S.

Cell Stem Cell. 2018 Dec 6;23(6):850-858.e4.

  • ヒトiPS細胞

Organoids from Nephrotic Disease-Derived iPSCs Identify Impaired NEPHRIN Localization and Slit Diaphragm Formation in Kidney Podocytes

Tanigawa S, Islam M, Sharmin S, Naganuma H, Yoshimura Y, Haque F, Era T, Nakazato H, Nakanishi K, Sakuma T, Yamamoto T, Kurihara H, Taguchi A, Nishinakamura R

Stem Cell Reports. 2018 Sep 11;11(3):727-740.

  • ラット毛包幹細胞

miR‑339‑5p negatively regulates loureirin A‑induced hair follicle stem cell differentiation by targeting DLX5.

Li X, Wu Y, Xie F, Zhang F, Zhang S, Zhou J, Chen D, Liu A

Mol Med Rep. 2018 Aug;18(2):1279-1286.

  • ヒトES細胞

Human Brain Organoids on a Chip Reveal the Physics of Folding

Karzbrun E, Kshirsagar A, Cohen SR, Hanna JH, Reiner O

Nat Phys. 2018 May;14(5):515-522.

  • ヒトiPS細胞

Generation of D1-1 TALEN isogenic control cell line from Dravet syndrome patient iPSCs using TALEN-mediated editing of the SCN1A gene.

Tanaka Y, Sone T, Higurashi N, Sakuma T, Suzuki S, Ishikawa M, Yamamoto T, Mitsui J, Tsuji H, Okano H, Hirose S

Stem Cell Res. 2018 Apr;28:100-104.

  • ヒトiPS細胞

Microhomology-assisted Scarless Genome Editing in Human iPSCs

Kim SI, Matsumoto T, Kagawa H, Nakamura M, Hirohata R, Ueno A, Ohishi M, Sakuma T, Soga T, Yamamoto T, Woltjen K.

Nat Commun. 2018 Mar 5;9(1):939.

  • ヒトiPS細胞

Down-regulation of Ghrelin Receptors on Dopaminergic Neurons in the Substantia Nigra Contributes to Parkinson's Disease-Like Motor Dysfunction

Suda Y, Kuzumaki N, Sone T, Narita M, Tanaka K, Hamada Y, Iwasawa C, Shibasaki M, Maekawa A, Matsuo M, Akamatsu W, Hattori N, Okano H, Narita M.

Mol Brain. 2018 Feb 20;11(1):6.

  • ヒトiPS細胞

Pluripotent stem cell models of Blau syndrome reveal an IFN-γ-dependent inflammatory response in macrophages.

Takada S, Kambe N, Kawasaki Y, Niwa A, Honda-Ozaki F, Kobayashi K, Osawa M, Nagahashi A, Semi K, Hotta A, Asaka I, Yamada Y, Nishikomori R, Heike T, Matsue H, Nakahata T, Saito MK

J Allergy Clin Immunol. 2018 Jan;141(1):339-349.e11.

  • ヒトiPS細胞

Site-specific Randomization of the Endogenous Genome by a Regulatable CRISPR-Cas9 piggyBac System in Human Cells

Ishida K, Xu H, Sasakawa N, Lung MSY, Kudryashev JA, Gee P, Hotta A.

Sci Rep. 2018 Jan 10;8(1):310.

  • ヒトiPS細胞

A human iPS cell myogenic differentiation system permitting high-throughput drug screening

Uchimura T, Otomo J, Sato M, Sakurai H.

Stem Cell Res. 2017 Dec:25:98-106.

  • ヒトiPS細胞

CRISPR Correction of a Homozygous Low-Density Lipoprotein Receptor Mutation in Familial Hypercholesterolemia Induced Pluripotent Stem Cells

Omer L, Hudson EA, Zheng S, Hoying JB, Shan Y, Boyd NL.

Hepatol Commun. 2017 Nov;1(9):886-898.

  • ヒト脂肪組織由来間葉系幹細胞（hAMSCs）

CRISPR/Cas9-Mediated Knockin Application in Cell Therapy: A Non-viral Procedure for Bystander Treatment of Glioma in Mice.

Meca-Cortés O, Guerra-Rebollo M, Garrido C, Borrós S, Rubio N, Blanco J.

Mol Ther Nucleic Acids. 2017 Sep 15:8:395-403.

  • ヒトiPS細胞

Impairment of the transition from proliferative stage to prehypertrophic stage in chondrogenic differentiation of human induced pluripotent stem cells harboring the causative mutation of achondroplasia in fibroblast growth factor receptor 3

Horie N, Hikita A, Nishizawa S, Uto S, Takato T, Hoshi K.

Regen Ther. 2017 Jan 26:6:15-20.

  • ヒトiPS細胞

Myogenic Differentiation from MYOGENIN-Mutated Human iPS Cells by CRISPR/Cas9.

Higashioka K, Koizumi N, Sakurai H, Sotozono C, Sato T

Stem Cells Int. 2017;2017:9210494.

  • ヒトiPS細胞

CRISPR/Cas9 Genome Editing to Repair Receptor-Mediated Endocytosis in Homozygous Familial Hypercholesterolemia Induced Pluripotent Stem Cells

Linda Omer, Elizabeth A Hudson, Jay B Hoying, Nolan L Boyd

The FASEB Journal Vol. 31, 909.2, 1 Apr 2017

  • ヒトiPS細胞

Proteasome Impairment in Neural Cells Derived From HMSN-P Patient iPSCs

Murakami N, Imamura K, Izumi Y, Egawa N, Tsukita K, Enami T, Yamamoto T, Kawarai T, Kaji R, Inoue H.

Mol Brain, 10 (1), 7 2017 Feb 15

  • トガリネズミ精原幹細胞

Long-term Propagation of Tree Shrew Spermatogonial Stem Cells in Culture and Successful Generation of Transgenic Offspring

Li CH, Yan LZ, Ban WZ, Tu Q, Wu Y, Wang L, Bi R, Ji S, Ma YH, Nie WH, Lv LB, Yao YG, Zhao XD, Zheng P.

Cell Res. 2017 Feb;27(2):241-252.

  • ヒトiPS細胞

Cochlear Cell Modeling Using Disease-Specific iPSCs Unveils a Degenerative Phenotype and Suggests Treatments for Congenital Progressive Hearing Loss.

Hosoya M, Fujioka M, Sone T, Okamoto S, Akamatsu W, Ukai H, Ueda HR, Ogawa K, Matsunaga T, Okano H

Cell Rep. 2017 Jan 3;18(1):68-81.

  • ヒトiPS細胞

Genetic and Pharmacological Correction of Aberrant Dopamine Synthesis Using Patient iPSCs With BH4 Metabolism Disorders

Ishikawa T, Imamura K, Kondo T, Koshiba Y, Hara S, Ichinose H, Furujo M, Kinoshita M, Oeda T, Takahashi J, Takahashi R, Inoue H.

Hum Mol Genet. 2016 Dec 1;25(23):5188-5197.

  • ヒトiPS細胞

Calcium Dysregulation Contributes to Neurodegeneration in FTLD Patient iPSC-derived Neurons

Imamura K, Sahara N, Kanaan NM, Tsukita K, Kondo T, Kutoku Y, Ohsawa Y, Sunada Y, Kawakami K, Hotta A, Yawata S, Watanabe D, Hasegawa M, Trojanowski JQ, Lee VM, Suhara T, Higuchi M, Inoue H

Sci Rep. 2016 Oct 10:6:34904.

  • ヒトiPS細胞

Human Induced Pluripotent Stem Cell-Derived Podocytes Mature Into Vascularized Glomeruli Upon Experimental Transplantation

Sharmin S, Taguchi A, Kaku Y, Yoshimura Y, Ohmori T, Sakuma T, Mukoyama M, Yamamoto T, Kurihara H, Nishinakamura R.

J Am Soc Nephrol. 2016 Jun;27(6):1778-91.

  • ヒトiPS細胞

Inducible Transgene Expression in Human iPS Cells Using Versatile All-in-One piggyBac Transposons.

Kim SI, Oceguera-Yanez F, Sakurai C, Nakagawa M, Yamanaka S, Woltjen K

Methods Mol Biol. 2016;1357:111-31.

  • ヒトiPS細胞

Sall1 transiently marks undifferentiated heart precursors and regulates their fate.

Morita Y, Andersen P, Hotta A, Tsukahara Y, Sasagawa N, Hayashida N, Koga C, Nishikawa M, Saga Y, Evans SM, Koshiba-Takeuchi K, Nishinakamura R, Yoshida Y, Kwon C, Takeuchi JK

J Mol Cell Cardiol. 2016 Mar;92:158-62.

  • ヒトiPS細胞

Efficient genomic correction methods in human iPS cells using CRISPR-Cas9 system.

Li HL, Gee P, Ishida K, Hotta A

Methods. 2015 Oct 23. pii: S1046-2023(15)30133-X.

  • ヒトiPS細胞

Targeted Gene Correction of RUNX1 in Induced Pluripotent Stem Cells Derived From Familial Platelet Disorder With Propensity to Myeloid Malignancy Restores Normal Megakaryopoiesis

Iizuka H, Kagoya Y, Kataoka K, Yoshimi A, Miyauchi M, Taoka K, Kumano K, Yamamoto T, Hotta A, Arai S, Kurokawa M.

Exp Hematol. 2015 Oct;43(10):849-57.

  • ヒトiPS細胞

Early pathogenesis of Duchenne muscular dystrophy modelled in patient-derived human induced pluripotent stem cells

Shoji E, Sakurai H, Nishino T, Nakahata T, Heike T, Awaya T, Fujii N, Manabe Y, Matsuo M, Sehara-Fujisawa A.

Sci Rep. 2015 Aug 20:5:12831.

  • ヒトES細胞

Gene Manipulation of Human Embryonic Stem Cells by In Vitro-Synthesized mRNA for Gene Therapy

Wang XL, Yu L, Ding Y, Guo XR, Yuan YH, Li DS

Curr Gene Ther. 2015;15(4):428-35.

  • ヒトiPS細胞

Precise Correction of the Dystrophin Gene in Duchenne Muscular Dystrophy Patient Induced Pluripotent Stem Cells by TALEN and CRISPR-Cas9.

Li HL, Fujimoto N, Sasakawa N, Shirai S, Ohkame T, Sakuma T, Tanaka M, Amano N, Watanabe A, Sakurai H, Yamamoto T, Yamanaka S, Hotta A.

Stem Cell Reports. 2015 Jan 13;4(1):143-154.

エレクトロポレーション

ドラッグデリバリー・遺伝子導入

電気式細胞融合

蛍光組織染色・in situ HCR

細胞分離

1細胞回収・マイクロダイセクション

細胞凍結

細胞･微生物培養 (解析/計数/伸展/灌流)

In vivo 超音波イメージング

卵振動培養