iPS Cells (iPSC from Fibroblasts, Episomal, Male, Control Lines for Genome Editing Services) *Academic Price

SKU :
ASE-9211
Catalog # :
ASE-9211
Size :
0.5 x 10^6 cells/vial
$2,500.00
Description

MTA required

Applied StemCell, Inc. provides Control Human Induced Pluripotent Stem (iPS) cells at low passages (p15). These pluripotent cells were generated from normal human skin fibroblasts using episomal reprogramming methods.  This method allows the transient expression of human transcription factors (OCT4, SOX2, KLF4, and c-MYC) that initiate the reprogramming process. The resulting human iPS cells (hiPSCs) were selected using morphological criteria without the use of either fluorescent markers or drug selection. These iPS cells have been tested for the expression of the pluripotency markers, including OCT4, SOX2, SSEA4, TRA-1-60, TRA-1-81, and alkaline phosphatase (AP) activity, and normal male karyotype. The ASE-9211 control human iPSC line can be used for CRISPR/Cas9 genome editing and differentiation to somatic lineages in vitro. Detailed protocols for thawing, culturing under feeder-free conditions, and cryopreservation of these iPS cells are provided.

For donor information, please refer to datasheet or contact us.


Characterization

Pluripotency Marker Analysis

ase-9211-control-ipsc-male-episomal-2 

Figure 1. Expression of pluripotency markers. ASE-9211 iPS cell line expresses common iPSC biomarkers (OCT4, SOX2, SSEA4, TRA-1-60, and TRA-1-81).  The corresponding DAPI staining is below each marker staining image.  All images were taken at 10x magnification.

Alkaline Phosphatase (AP) Staining

ase-9211-control-ipsc-episomal-male-3 

Figure 2. ASE-9211 iPSCs stain positive for Alkaline Phosphatase: (a) A typical unstained colony (a) was used to gauge the extent of the AP staining (b). Both images were taken at 5x magnification.

Karyotype Analysis

 landingpage-ase-9211-karyotype-5   landingpage-ase-9211-karyotype-results

Figure 3. Cytogenic analysis was performed on G-banded metaphase cells from human iPSC line, ASE-9211 at passage 15. This iPSC line demonstrates a normal male karyotype.

Karyotype Analysis

    landingpage-ase-9211-genetic-analysislandingpage-ase-9211-genetic-analysis-1

Figure 4. Quantitative PCR (qPCR) analysis to detect karyotypic abnormalities in control human iPSC line, ASE-9211 shows a normal male karyotype in all loci tested.


CRISPR Genome Engineering Capability

 

landingpage-ase-9211-crispr-genome-editing

Figure 5. CRISPR/Cas9 was used to generate 6 knockout cell lines from the control human iPSC line, ASE-9211.


Differentiation Capability

Control iPSC line, ASE-9211 was used for differentiation into motor neurons using proprietary, integration-free protocols. 

appnote-service-stemcell-nscdiff-motorneuron-1


Figure 6. Immunocytochemical staining images of the motor neuron derived from control "master" iPSC line, ASE-9211 at 2 days post thaw. The iPSC-derived motor neurons were stained with antibodies for motor neuron biomarker, HB9 (green), and neuronal biomarker, Tuj1 (red). DAPI (blue) was used as nucleus stain.

appnote-service-stemcell-nscdiff-motorneuron-2

Figure 7. Immunocytochemical staining for motor neuron marker, ChAT (green) and neuronal marker, MAP2 i(blue) n motor neurons differentiated from control "master" iPSC line, ASE-9211 at 7 days post thaw. DAPI (blue) was used for nucleus staining.