SKIP000203 | |
SKIP ID | SKIP000203 |
Organism(En) | - |
Organism(Ja) | - |
Cell Type(En) | - |
Cell Type(Ja) | - |
Cell Tissue(En) | - |
Cell Tissue(Ja) | - |
Cell Origin | Diseased |
Cell Name 1(En) | PB2 |
Cell Name 1(Ja) | PB2 |
Cell Name 2(En) | - |
Cell Name 2(Ja) | - |
Disease Name 1(Ja) | 家族性パーキンソン病 |
ICD Code 1 | G20 |
Disease Name 1(En) | Parkinson disease |
OMIM1 | 600116 |
Disease Name 2(Ja) | - |
ICD Code 2 | - |
Disease Name 2(En) | - |
OMIM 2 | - |
Disease Name 3(Ja) | - |
ICD Code 3 | - |
Disease Name 3(En) | - |
OMIM 3 | - |
Age | - |
Age Range | 50-59 |
Sex | Male |
Race(En) | - |
Race(Ja) | - |
Genetic Diagnosis | Yes |
Not Detected | No |
Description(En) | PARK2 iPSC, Exon 6,7 homozygous deletions in parkin |
Description(Ja) | PARK2 ヒト iPS細胞, parkin遺伝子Exon 6,7 両染色体欠失 |
Cell Morphology | human ES-like |
Grade | -- |
Vector | Retrovirus |
Transgene | Retrovirus pMXs Oct3/4, Sox2, Klf4, c-Myc |
Adhesiveness | Yes |
Feeder | Yes |
Feeder Cell | SNL 1.5x10^(6)/10cm dish, Mitomycin C treatment:400microG/ml, 2hr 15min |
Medium | DMEM/F12(SIGMA; D6421) |
Genome Editing | - |
CO2 | - |
Mycoplasma | - |
Detection of Contaminants Mycoplasma | - |
Pluripotent Markers | - |
Pluripotent Markers Assay | - |
in vitro Differentiation | - |
in vitro Differentiation Assay | - |
in vivo Differentiation | - |
in vivo Differentiation Assay | - |
Other 1 Assay | - |
Other 1 Assay Method | - |
Other 2 Assay | - |
Other 2 Assay Method | - |
Other 3 Assay | - |
Other 3 Assay Method | - |
Karyotype | - |
Karyotype Assay | - |
Remaining Vector Detection | - |
Remaining Vector Detection Assay | - |
STR | - |
HLA | - |
Stem Cell Transcriptome analysis | - |
Stem Cell Transcriptome analysis Assay | - |
Author Name(En) | Hideyuki Okano |
Author Name(Ja) | 岡野 栄之 |
Author Organization(En) | Keio University |
Author Organization(Ja) | 慶應義塾大学 |
Author Contact Email | - |
PI Organization(En) | Keio University |
PI Organization(Ja) | 慶應義塾大学 |
PI Name(En) | Hideyuki Okano |
PI Name(Ja) | 岡野 栄之 |
PI Contact Email | - |
Availability | Available |
Provider Organization(En) | Keio University |
Provider Organization(Ja) | 慶應義塾大学医学部 |
Provider Email | akamatsu[at]a7[dot]keio[dot]jp |
Provider URL | - |
Ethical Statement(En) | - |
Ethical Statement(Ja) | - |
Terms of Use(En) | - |
Terms of Use(Ja) | - |
PubMed ID | 23039195 |
DOI | 10.1186/1756-6606-5-35 |
Title | Mitochondrial dysfunction associated with increased oxidative stress and alpha-synuclein accumulation in PARK2 iPSC-derived neurons and postmortem brain tissue. |
Authors | Imaizumi Y, Okada Y, Akamatsu W, Koike M, Kuzumaki N, Hayakawa H, Nihira T, Kobayashi T, Ohyama M, Sato S, Takanashi M, Funayama M, Hirayama A, Soga T, Hishiki T, Suematsu M, Yagi T, Ito D, Kosakai A, Hayashi K, Shouji M, Nakanishi A, Suzuki N, Mizuno Y, Mizushima N, Amagai M, Uchiyama Y, Mochizuki H, Hattori N, Okano H |
Journal | Mol Brain |
Year | 2012 |
Volume | 5 |
Issue | - |
Pages | 35 |
URL | http://www.ncbi.nlm.nih.gov/pubmed/?term=23039195 |
Free input | - |
Note | 確定診断方法:parkin遺伝子のexon毎に設計したprimerを用いてgenomic PCR解析を行った。その結果、 PB1,2,18,20iPSにおいてExon 6, 7のhomozygous deletionを確認。 HIV : 未実施 HTLV-1 : 未実施 HBV : 実施 陰性 HCV : 実施 陰性 病歴・治療歴等 : 28歳発症 家族歴 : 両親に血族結婚あり 飲酒・喫煙の嗜好歴 : 維持培養方法 1.基本培地 DMEM/F12(SIGMA; D6421) 2.血清: 使用しない 3.添加物: 使用する KockOutTM Serum Replecement(GIBCO; 10828-028) 20% Non Essential Amino Acid Solution(SIGMA; M7145) 10% b-mercaptoethanol(SIGMA; M7522) 0.1% 200mM L-Glutamine(Nacalai tesque; 16948-04) 1% human-FGF basic(PEPRO TECH; 100-18B)4ng/mL 4.抗生物質(含抗真菌剤) : 使用する Penicillin/Streptomycin(Nacalai tesque; 26253-84) 0.5% 5.培養温度 : 37℃ 6.CO2濃度 : 3.0% 7.培養容器 : 10 cm dish(日本ジェネティクス; FG-2090) コーティングの有無 : コーティングする Gelatin from porcine skin Type A(SIGMA; G2625) 8.Feeder Cellの使用: 使用する 細胞名 : SNL細胞 使用時の細胞密度 : 1.5 x 10^(6)/10cm dish 使用前処理方法 : Mitomycin C処理 : 400microG/mL, 2hr 15min 9.継代方法 [1]Aspirate medium. [2]Wash once with PBS. [3]Add Dissociation solution 1mL. [4]Aspirate Dissociation solution. [5]Incubate 7min at 37 degree. [6]Add 6ml Medium and collect colonies by mild pipetting. [7]Spin down at 160g 5min. [8]Aspirate supernatant and add appropriate volume of hiPS medium. [9]Dissociate colonies by Pipetting 5-6 times with 10ml pipette.(dissociate into small cell clamps) [10]Passage the cells 1:3-1:8. Culture at 37 degree, 3% CO2. *10mL plastic disposable pipette(greiner; 607160) *Dissociation solution 2.5% Trypsin 5ml(GIBCO; 15090-046) 1mg/mL Coll IV 5ml(GIBCO; 17104-019) 1M CaCl2 0.05ml(Nacalai tesque; 06729-55) KSR 10ml(GIBCO; 10828-028) Total 50ml Aliquot and store at -20 degree 10.パッセージの際の細胞播種時の細胞密度 Confluentの10cm dish 1枚から、10cm dish3〜8枚にパッセージ 11.継代頻度 : コロニーが分化する直前 コロニーの中心および辺縁が茶色くなり始めた時点 4-6日に一回 12.その他 凍結保存方法 ・ガラス化法 [1]Wash cells once with PBS, add and aspirate dissociation solution, incubate 7min at 37 degree, and add 6mL of hiPS medium per 10cm dish. [2]Carefully detach colonies with cell scraper(keep original colony size). [3]Collect medium + colonies using 5mL pipette(grainer-important point). [4]Divide 6mL colony suspension into 2mL x 3 tupes(15mL falcons) and spin at 160 g for 5min at 4 degree. [5]Aspirate medium and put tubes with peletts on ice. Freezing procedure: [6]Take 200 microL of freezing solution using a P1000 pipette, and reset the dial to around 300 microL. [7]Resuspend pellet and pipette once or twice and immediately transfer to cryotube. [8]Immediately transfer cryotube directly to flask containing liquid N2 for 15s abd drio the tupe into liquid N2. [9]Steps 8-9 must be performed within 15 seconds. ・Freezing solution(DAP213) [1]0.59g 1M acetamide to 6mL of hiPS medium to 15ml tube [2]Add 1.42mL 2M DMSO & 2.2mL Propylene glycol [3]Measure up to the 10mL line by adding hiPS medium Sterile filter through 0.2microM filters, Store at -80℃ Acetamide(SIGMA;A0500) Dimethyl Sulfoxide(SIGMA;D5879) Propylene glycol(SIGMA;398039) 0.2microM filter(Millipore;SLGV033RB) 継代数 : PB1(P10), PB2(P9), PB18(P9), PB20(P6) 確認済の未分化マーカー Oct3/4 : qPCR法、ICC法 Nanog : ICC法 SSEA-4 : ICC法 Tra-1-60 : ICC法 Tra-1-81 : ICC法 染色体解析 : 実施 CGH array解析済み。PARK2遺伝子の欠損以外、変異は見当たらなかった。 胚葉体形成実験 : 実施 胚葉体形成確認 テラトーマ形成実験 : 実施 テラトーマ形成確認 in vitro分化能解析 : 実施 神経への分化確認 マイコプラズマ汚染検査 : 未実施 細胞同定検査(STR多型解析) : 未実施 クローニング : 未実施 |