Cell Line



BCRC Number: SC81042
Name: IBMS-iPSC-012-12
Type: Sendai virus reprogrammed human induced pluripotent stem cell
Species: Homo sapiens, Human
Strain Ethnicity: Asian
Age Stage: 35 years old
Gender: Male
Tissue: Peripheral blood mononuclear cell (PBMC)

Morphology: Colonies with smooth surfaces and well defined edges, ES-like
Split Ratio: Depend on culture vessel (10~20 colonies per 6cm dish)
Mycoplasma Test: Negative

Incubation Temp(°C): 37°C
Incubation Condition: 5% CO2 incubator
Culture Medium: 80% DMEM/F12 with 2mM L-glutamine adjusted to contain 1mM sodium pyruvate + 1% NEAA + 1% GlutaMAX + 55uM 2-mercaptoethanol + 1% Penicillin-Streptomycin + 10ng/ml bFGF + 20% Knockout serum replacement (Prepare the iPSC medium without bFGF, and then supplement with fresh bFGF when the medium is used.)
Growth Property: Growth as colony-type
SubCulture Procedure: MEF based culture systems, FIRDI recommends plating density for inactivated MEFs is 1x10^6 cells per 10 cm dish (3.33x10^5 cells 6 cm dish).
Day0
Preparing MEF Culture Dishes
1. Cover the whole surface of each culture vessel with 0.1% gelatin and incubate the vessels for 30 minutes at 37°C or for 2 hours at room temperature.
2. Aspirate the gelatin solution from the gelatin coated culture vessel.
3. Into each of these culture vessels, add the appropriate amount of MEF suspension.
4. Incubate the cells in a 37°C incubator with a humidified atmosphere of 5% CO2.
Day1
There are two methods for iPSCs passaging:
Method A. Passaging iPSCs Using mechanical passage:
1. Pre-warm iPSC Medium and MEF culture vessel Ready.
2. The prepared MEF culture vessel for culture by aspirating the medium from the vessel, and rinsing with sterile DPBS, add complete iPSC Medium to MEF culture vessel.
3. Aspirate the spent medium from the vessel containing iPSCs and add complete iPSC Medium (pre-warm iPSC medium with b-FGF).
4. Transfer the culture dish to a sterile cell culture hood equipped with a stereomicroscope.
5. Using a glass knife, cut the colony to be picked into 5–6 pieces in a grid-like pattern.
6. Using a 200μL pipette, transfer the cut pieces to a freshly prepared MEF culture vessel containing iPSC medium.
7. Incubate the MEF-culture plate containing the picked colonies in a 37°C incubator with a humidified atmosphere of 5% CO2.
8. Change the medium every day.
Method B. Passaging iPSCs Using Chemical passage:
1. Pre-warm iPSC Medium and MEF culture vessel Ready.
2. The prepared MEF culture vessel for culture by aspirating the medium from the vessel, and rinsing with sterile DPBS, add complete iPSC Medium to MEF culture vessel.
3. Aspirate the spent medium from the vessel containing iPSCs and add Accutase for 2min treatment.
4. Remove Accutase, add DPBS wash then aspirate.
5. Add complete iPSC Medium (pre-warm iPSC medium with b-FGF).
6. Use cell scraper cut the iPSCs to be picked into small pieces in a grid-like pattern.
7. Final transfer the iPSC pieces to the mouse embryonic fibroblasts (MEF) culture vessel containing iPSC medium.
8. Incubate the MEF-culture plate containing the picked colonies in a 37°C incubator with a humidified atmosphere of 5% CO2.
9. Change the medium every day.
Freezed Medium: DMEM/F12 + 25% Knockout serum replacement + 10% DMSO
Seeding Density: Depend on culture vessel (10~20 colonies per 6cm dish)
Medium Renewal: Every day

Deposit History: Academia Sinica - IBMS
Description: Disease iPSC : Polycystic Kidney Disease (PKD)
Others: Reference : Huang, C.Y., et al., Generation of induced pluripotent stem cells derived from an autosomal dominant polycystic kidney disease patient with a p.Ser1457fs mutation in PKD1. Stem Cell Res, 2017. 24: p. 139-143.
Category: disease iPS

Image / Description:   細胞解凍後14天_40X
 

Biosafety Level: 2
Use Restriction: After agreement of depositor, research use only
Sponsor: MOST-NCFB
Shipping/Field/Price:
Frozen Vial for academic NTD$ 10000 Order this item
Frozen Vial for industry NTD$ 30000 Order this item

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