SKIP000768 | |
SKIP ID | SKIP000768 |
Organism(En) | - |
Organism(Ja) | - |
Cell Type(En) | - |
Cell Type(Ja) | - |
Cell Tissue(En) | - |
Cell Tissue(Ja) | - |
Cell Origin | Diseased |
Cell Name 1(En) | SLS-1 |
Cell Name 1(Ja) | SLS-1 |
Cell Name 2(En) | - |
Cell Name 2(Ja) | - |
Disease Name 1(Ja) | シェーグレン・ラルソン症候群 |
ICD Code 1 | Q871 |
Disease Name 1(En) | Sjogren-Larsson syndrome |
OMIM1 | 270200 |
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 | -- |
Sex | -- |
Race(En) | - |
Race(Ja) | - |
Genetic Diagnosis | Yes |
Not Detected | No |
Description(En) | iPS cell line derived from a patient with Sjogren-Larsson syndrome(ALDH3A2 1339A>G(K447E) and c.57-132 dup fs 218X) |
Description(Ja) | シェーグレン・ラルソン症候群患者由来iPS細胞株(ALDH3A2遺伝子変異1339A>G(K447E)およびc.57-132 dup fs 218X) |
Cell Morphology | human ES-like |
Grade | Research Grade |
Vector | Retrovirus |
Transgene | Retrovirus pMXs Oct3/4, Sox2, Klf4, c-Myc |
Adhesiveness | Yes |
Feeder | Yes |
Feeder Cell | SNL 1.5x10^(6)/10cm dish |
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 | Information Only |
Provider Organization(En) | - |
Provider Organization(Ja) | - |
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 | - |
DOI | - |
Title | - |
Authors | - |
Journal | - |
Year | - |
Volume | - |
Issue | - |
Pages | - |
URL | - |
Free input | - |
Note | 確定診断方法:ALDH3A2遺伝子変異 1339A>G(K447E)およびc.57-132 dup fs218X 血液サンプルからゲノムDNAを抽出、これを鋳型とし、ALDH3A2 遺伝子 (GenBank accession NoNM_000382)中の翻訳領域をPCR法で増幅し、 増幅産物をダイレクトシークエンス法により分析した。 HIV : 不明 HTLV-1 : 不明 HBV : 不明 HCV : 不明 病歴・治療歴等 : その他の既往歴 : 家族歴 : 飲酒・喫煙の嗜好歴 : 喫煙なし、飲酒なし 身体検査所見 : 臨床検査データ : 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 dish 3〜8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 : 確認済の未分化マーカー Oct3/4 : qPCR法 Nanog : qPCR法 染色体解析 : 未実施 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 : 未実施 in vitro分化能解析 : 未実施 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 その他の特性情報 : |