induced Pluripotent Stem Cell Core

100ug/mL stock, R&D Systems Cat# 338-AC-01M/CF

Gibco Cat#17504-044

Gibco Cat#A1895601

This service provides thawing, expansion and banking of iPSC lines derived by the Core or external laboratories under feeder-dependent or feeder-free conditions.

Prerequisites:

• Designation of iPSC line to be banked
• iPSC lines new to the facility will need to produce evidence of being mycoplasma-free

This service takes 2-5 weeks to complete and includes the following:

• Thaw cells on mouse feeders or under feeder-free conditions

• Transition cells from feeder-dependent culture to feeder-free conditions if requested
• Expansion of cells
• Cryopreservation of expanded cells into 20 vials per cell line
• Mycoplasma testing
• Transfer of all frozen vials

Additional Notes:

• If thawing fails, PI will be notified immediately

This service provides thawing, expansion and banking of iPSC lines derived by the Core or external laboratories under feeder-dependent or feeder-free conditions.

Prerequisites:

• Designation of iPSC line to be banked
• iPSC lines new to the facility will need to produce evidence of being mycoplasma-free

This service takes 2-5 weeks to complete and includes the following:

• Thaw cells on mouse feeders or under feeder-free conditions

• Transition cells from feeder-dependent culture to feeder-free conditions if requested
• Expansion of cells
• Cryopreservation of expanded cells into 20 vials per cell line
• Mycoplasma testing
• Transfer of all frozen vials

Additional Notes:

• If thawing fails, PI will be notified immediately

This service provides thawing, expansion and banking of iPSC lines derived by the Core or external laboratories under feeder-dependent or feeder-free conditions.

Prerequisites:

• Designation of iPSC line to be banked
• iPSC lines new to the facility will need to produce evidence of being mycoplasma-free

This service takes 2-5 weeks to complete and includes the following:

• Thaw cells on mouse feeders or under feeder-free conditions

• Transition cells from feeder-dependent culture to feeder-free conditions if requested
• Expansion of cells
• Cryopreservation of expanded cells into 20 vials per cell line
• Mycoplasma testing
• Transfer of all frozen vials

Additional Notes:

• If thawing fails, PI will be notified immediately

100ug/mL stock, PeproTech Cat#AF-120-05ET

R&D Systems cat no. 3433-005-01

Current Lot Protein Concentration: 8.32 mg/mL

This Service provides the design and the construction (cloning) of a plasmid template used for targeting iPSCs using homology directed repair (e.g. template for generating a gene reporter)

Prerequisites:

• Selection of a hESC or hiPSC line to be gene edited
• A Service Agreement must be executed for non-Penn users
• MTA must be executed if transferring a cell line to the Core
• All cell lines not generated by the Core must be shown evidence of being mycoplasma- free

This service takes 3-5 weeks to complete and includes the following:

• Consultation with the user to determine targeting strategy
• Design and construction of the donor vector
• Delivery of the donor vector as a DNA maxiprep

This service is normally requested in order to establish fibroblasts for iPSC reprogramming. 

This service takes 3-4 weeks to complete and includes the following:

• Fibroblast isolation from freshly collected skin biopsies
• Establishment of fibroblast culture
• Mycoplasma testing
• Cryopreservation of several vials of early passage cells
• Freezing of cell pellets for future genomic DNA isolation

This service establishes genome modified hESCs/hiPSCs using CRISPR/Cas9 technology

Prerequisites:

• Selection of a hESC or hiPSC line to be gene edited
• A Service Agreement must be executed for non-Penn users
• MTA must be executed if transferring a cell line to the Core
• All cell lines not generated by the Core must be shown evidence of being mycoplasma- free

This service takes 6-9 months to complete and includes the following:

• Consultation with the user to determine targeting strategy
• Design and construction of CRISPR gRNA(s) for targeting
• Validation of gRNA(s) in the targeting cell line
• Targeting of the cell line using validated gRNA(s)
• Screening of targeted clones
• Establishment of at least 2 correctly targeted clones
• Mycoplasma testing of the clones
• qPCR-based analysis for common karyotypic abnormalities of the clones
• Delivery of cryopreserved cells and written report

Prerequisites:

• Students must have at least one year of hands-on experience in mammalian cell culture

This is a hands-on training course in basic techniques in hESC/iPSC culture including cell thawing, expansion, identification by morphology, manual and enzymatic passaging, freezing, and feeder-free culture.  This course will take 2 weeks to complete (1-2 hours per day)

Under construction.  Please contact the Core director.

Under construction.  Please contact the Core director.

Prerequisites:

• All required IRB approval must be in place. Please submit IRB approval document here:
• A Service Agreement must be completed for non-Penn users
• Users should request the PBMC isolation from blood service or submit previously isolated PBMCs.
• If submitting PBMCs, evidence showing their mycoplasma free status must be shown before they are accepted.

This service takes 4-6 months to complete and includes the following:

• Culture of PBMCs for enrichment of erythroid progenitors
• Transduction of cultured cells with Sendai viral reprogramming vectors
• iPSC colony picking
• Expansion of iPSC clones in either feeder-dependent or feeder-free conditions
• Mycoplasma testing of expanded clones
• FACS analysis of 2 pluripotency markers
• qPCR-based analysis for common karyotypic abnormalities in iPSCs
• PCR assay for loss of Sendai viral reprogramming vectors
• Cryopreservation of iPSC lines (4 vials/clone)
• Will deliver 3 clones at passage ≥10

 ** 10% Discount for simulatneous reprogramming of 4 or more samples **

Check the box for either feeder-dependent or feeder-free culture for iPSC expansion

Prerequisites:

• All required IRB approval must be in place. 
• A Service Agreement must be completed for non-Penn users
• The Core accepts either live or frozen cell stocks.
• Submitted cells MUST be mycoplasma free before they are accepted.
• Cells should also be at low passage (preferably <5) and are proliferating as non-dividing cells are refractory to reprogramming.

This service takes 4-6 months to complete and includes the following:

• Expansion, mycoplasma testing, cryopreservation of fibroblast culture
• Transduction of fibroblasts with Sendai viral reprogramming vectors
• iPSC colony picking
• Expansion of iPSC clones in either feeder-dependent or feeder-free conditions
• Mycoplasma testing of expanded clones
• FACS analysis of 2 pluripotency markers
• qPCR-based analysis for common karyotypic abnormalities in iPSCs
• PCR assay for loss of Sendai viral reprogramming vectors
• Cryopreservation of iPSC lines (4 vials/clone)
• Will deliver 3 clones at passage ≥10

** 10% Discount for simulatneous reprogramming of 4 or more samples **

Prerequisites:

• iPS Cell line must be generated by the Core
• When submitting a non-iPSC line (i.e. parental cell line for iPSC derivation), user must submit a frozen cell pellet

This service will take 2-3 weeks to complete and includes the following:

• Expansion of iPSC line
• Freezing down of a cell pellet
• Submission of frozen cell pellet to a commercial vendor for STR analysis
• Delivery of STR analysis report

The Core has a number of wild-type control iPSC lines available to users.   Please inquire with the Core director at wenliyan@pennmedicine.upenn.edu.

This service can only be added in addition to iPSC-Derived Cardiomyocytes

This service provides iPSC-derived cardiomyocytes in monolayer cultures.  iPSCs are differentiated using cytokines and small molecules in defined media.  Cultures are then metabolically selected to enrich for cardiomyocytes and re-plated. These cells resemble immature cardiomyocytes and but also display a number of properties of adult cardiomyocytes. 

Prerequisites:

• Designation of iPSC line to be differentiated
• New iPSC lines will need to show evidence of being mycoplasma-free or this will be performed by the Core at extra charge (see Mycoplasma Testing service)

This service takes 5-8 weeks to complete and includes the following:

• Thaw cells and expand for differentiation
• Up to two rounds of pilot studies to determine optimal seeding density and conditions for differentiation 
• Directed differentiation in monolayer cultures
• Transfer of live monolayer cultures of beating cells in two 10-cm plates
• Approximate yield: 12-20 million cells

Additional Notes:

• Additional plates can be purchased at a reduced price.
• If differentiation fails in both pilot studies, PI will be notified immediately and the project will be discontinued.  Service fee still applies.
• More pilot studies can be performed using a different CM differentiation protocol after submitting a new request.

This service provides iPSC-derived cardiomyocytes in monolayer cultures.  iPSCs are differentiated using cytokines and small molecules in defined media.  Cultures are then metabolically selected to enrich for cardiomyocytes and re-plated. These cells resemble immature cardiomyocytes and but also display a number of properties of adult cardiomyocytes.  

Prerequisites:

• Designation of iPSC line to be differentiated

This service takes 5-8 weeks to complete and includes the following: 

• Thaw cells and expand for differentiation
• Directed differentiation in monolayer cultures using SOP established by the Core
• Transfer of live monolayer cultures of beating cells in two 10-cm plates
• Approximate yield: 12-20 million cells

 Additional Notes:

• Additional plates can be purchased at a reduced price

This service provides iPSC-derived endothelial cell in monolayer cultures.  iPSCs undergo directed differentiation using cytokines and small molecules to endothelial progenitors.   Cultures are then purified with endothelial specific cell surface markers to enrich for purity and further passaged.  These cells express CD31(PECAM) & CD144 (VE-cadherin) surface markers, uptake Ac-LDL, and form vessels when cultured with stromal cells in 3D cultures.

Prerequisites:

• Designation of iPS line to be differentiated
• New iPSC lines will need to show evidence of being mycoplasma-free

This service takes 4-6 weeks to complete and includes the following:

• Thaw cells and expand for differentiation
• Directed differentiation in monolayer cultures
• Magnetic activated cell sorting against endothelial markers
• Passaged in endothelial growth media for cell expansion
• Flow cytometry analysis of endothelial markers CD31/CD144
• Passage 3 live culture supplied in at least 2 6-well plates
• Approximate yield: 6 million cells
• Guaranteed purity > 90% CD31⁺/CD144⁺

Additional Notes:

• If differentiation fails, PI will be notified immediately
• Additional plates can be purchased at a reduced price

This service establishes genome modified hESCs/hiPSCs using CRISPR/Cas9 technology

Prerequisites:

• Selection of a hESC or hiPSC line to be gene edited
• A Service Agreement must be executed for non-Penn users
• MTA must be executed if transferring a cell line to the Core
• All cell lines not generated by the Core must be shown evidence of being mycoplasma- free

This service takes 6-9 months to complete and includes the following:

• Consultation with the user to determine targeting strategy
• Design and construction of CRISPR gRNA(s) for targeting
• Validation of gRNA(s) in the targeting cell line
• Targeting of the cell line using validated gRNA(s) and HDR DNA donor template
• Screening of targeted clones
• Establishment of at least 2 correctly targeted clones
• Mycoplasma testing of the clones
• qPCR-based analysis for common karyotypic abnormalities of the clones
• Delivery of cryopreserved cells and written report

This service establishes genome modified hESCs/hiPSCs using CRISPR/Cas9 technology

Prerequisites:

• Selection of a hESC or hiPSC line to be gene edited
• A Service Agreement must be executed for non-Penn users
• MTA must be executed if transferring a cell line to the Core
• All cell lines not generated by the Core must be shown evidence of being mycoplasma-free

This service takes 6-9 months to complete and includes the following:

• Consultation with the user to determine targeting strategy
• Design and construction of CRISPR gRNA(s) for targeting
• Validation of gRNA(s) in the targeting cell line
• Design and synthesis of the ssODN donor template
• Targeting of the cell line using validated gRNA(s) and ssODN donor template
• Screening of targeted clones
• Establishment of at least 2 correctly targeted clones
• Mycoplasma testing of the clones
• qPCR-based analysis for common karyotypic abnormalities of the clones
• Delivery of cryopreserved cells and written report

Gibco Catalog # 10828028

We provide limited tissue culture space to UPenn investigators who elect to perform their experiments in the iPSC core with guidance from the Core staff.   Investigators have full access to our hESC/iPSC dedicated mycoplasma-free cell culture rooms, consumables and some commonly used stem cell culture reagents, such as PBS and DMEM/F12.   All other reagents, such as stem cell medium, coating matrix are charged separately or provided by the user.

Prerequisites:

• Must have completed the hESC/hiPSC culture hands-on training course
• Must have approval from Core director following a consultation meeting

Stem Cell Technologies Catalog # 100-0276

Prerequisites:

• Cell line must be generated by the Core

This service takes 1 week to complete and includes the following:

• Isolation of total RNA from iPSC line
• qRT-PCR analysis of 5 stem cell marker genes
• Gene expression of parental cells and a previously established iPSC line included as controls
• Will deliver analysis report

Prerequisites:

• Cell line must be generated by the Core

This service will take 3-4 weeks to complete and includes the following:

• Expansion and plating of iPSC line for tri-lineage differentiation
• Directed differentiation of iPSCs to mesoderm, endoderm and ectoderm lineages separately
• Flow cytometry analysis of 2 marker proteins per cell lineage
• Will deliver analysis report

This service is normally requested in order to isolate PBMCs from blood draws for iPSC reprogramming.  Freshly isolated cells can be reprogrammed immediately or can be revived at a later time for iPSC reprogramming.  This service takes 1 day to complete and includes the following:

• Isolation of PBMCs from blood using SEPMate (Stem Cell Technologies)
• Cryopreservation of several vials of PBMCs (1-3 million per vial) 

Note:  This service does not include storage of cryopreserved vials in the Core facility.

100ug/mL stock, R&D Systems Cat#233-FB-001MG/CF

Miltenyi Biotec Catolog #130-104-368

Prerequisites:

• All required IRB approval must be in place. Please submit IRB approval document here:
• A Service Agreement must be completed for non-Penn users
• Users should request the PBMC isolation from blood service or submit previously isolated PBMCs.
• If submitting PBMCs, evidence showing their mycoplasma free status must be shown before they are accepted.

This service takes 4-6 months to complete and includes the following:

• Culture of PBMCs for enrichment of erythroid progenitors
• Transduction of cultured cells with Sendai viral reprogramming vectors
• iPSC colony picking
• Expansion of iPSC clones in either feeder-dependent or feeder-free conditions
• Mycoplasma testing of expanded clones
• FACS analysis of 2 pluripotency markers
• qPCR-based analysis for common karyotypic abnormalities in iPSCs
• PCR assay for loss of Sendai viral reprogramming vectors
• Cryopreservation of iPSC lines (4 vials/clone)
• Will deliver 3 clones at passage ≥10

 ** 10% Discount for simulatneous reprogramming of 4 or more samples **

Check the box for either feeder-dependent or feeder-free culture for iPSC expansion

Prerequisites:

• All required IRB approval must be in place. 
• A Service Agreement must be completed for non-Penn users
• The Core accepts either live or frozen cell stocks.
• Submitted cells MUST be mycoplasma free before they are accepted.
• Cells should also be at low passage (preferably <5) and are proliferating as non-dividing cells are refractory to reprogramming.

This service takes 4-6 months to complete and includes the following:

• Expansion, mycoplasma testing, cryopreservation of fibroblast culture
• Transduction of fibroblasts with Sendai viral reprogramming vectors
• iPSC colony picking
• Expansion of iPSC clones in either feeder-dependent or feeder-free conditions
• Mycoplasma testing of expanded clones
• FACS analysis of 2 pluripotency markers
• qPCR-based analysis for common karyotypic abnormalities in iPSCs
• PCR assay for loss of Sendai viral reprogramming vectors
• Cryopreservation of iPSC lines (4 vials/clone)
• Will deliver 3 clones at passage ≥10

** 10% Discount for simulatneous reprogramming of 4 or more samples **

This service is normally requested in order to isolate PBMCs from blood draws for iPSC reprogramming.  Freshly isolated cells can be reprogrammed immediately or can be revived at a later time for iPSC reprogramming.  This service takes 1 day to complete and includes the following:

• Isolation of PBMCs from blood using SEPMate (Stem Cell Technologies)
• Cryopreservation of several vials of PBMCs (1-3 million per vial) 

Note:  This service does not include storage of cryopreserved vials in the Core facility.

This service is normally requested in order to establish fibroblasts for iPSC reprogramming. 

This service takes 3-4 weeks to complete and includes the following:

• Fibroblast isolation from freshly collected skin biopsies
• Establishment of fibroblast culture
• Mycoplasma testing
• Cryopreservation of several vials of early passage cells
• Freezing of cell pellets for future genomic DNA isolation

Prerequisites:

• Cell line must be generated by the Core

This service takes 1 week to complete and includes the following:

• Isolation of total RNA from iPSC line
• qRT-PCR analysis of 5 stem cell marker genes
• Gene expression of parental cells and a previously established iPSC line included as controls
• Will deliver analysis report

Prerequisites:

• Cell line must be generated by the Core

This service will take 3-4 weeks to complete and includes the following:

• Expansion and plating of iPSC line for tri-lineage differentiation
• Directed differentiation of iPSCs to mesoderm, endoderm and ectoderm lineages separately
• Flow cytometry analysis of 2 marker proteins per cell lineage
• Will deliver analysis report

Prerequisites:

• iPS Cell line must be generated by the Core
• When submitting a non-iPSC line (i.e. parental cell line for iPSC derivation), user must submit a frozen cell pellet

This service will take 2-3 weeks to complete and includes the following:

• Expansion of iPSC line
• Freezing down of a cell pellet
• Submission of frozen cell pellet to a commercial vendor for STR analysis
• Delivery of STR analysis report

This service provides thawing, expansion and banking of iPSC lines derived by the Core or external laboratories under feeder-dependent or feeder-free conditions.

Prerequisites:

• Designation of iPSC line to be banked
• iPSC lines new to the facility will need to produce evidence of being mycoplasma-free

This service takes 2-5 weeks to complete and includes the following:

• Thaw cells on mouse feeders or under feeder-free conditions

• Transition cells from feeder-dependent culture to feeder-free conditions if requested
• Expansion of cells
• Cryopreservation of expanded cells into 20 vials per cell line
• Mycoplasma testing
• Transfer of all frozen vials

Additional Notes:

• If thawing fails, PI will be notified immediately

This service provides thawing, expansion and banking of iPSC lines derived by the Core or external laboratories under feeder-dependent or feeder-free conditions.

Prerequisites:

• Designation of iPSC line to be banked
• iPSC lines new to the facility will need to produce evidence of being mycoplasma-free

This service takes 2-5 weeks to complete and includes the following:

• Thaw cells on mouse feeders or under feeder-free conditions

• Transition cells from feeder-dependent culture to feeder-free conditions if requested
• Expansion of cells
• Cryopreservation of expanded cells into 20 vials per cell line
• Mycoplasma testing
• Transfer of all frozen vials

Additional Notes:

• If thawing fails, PI will be notified immediately

This service provides thawing, expansion and banking of iPSC lines derived by the Core or external laboratories under feeder-dependent or feeder-free conditions.

Prerequisites:

• Designation of iPSC line to be banked
• iPSC lines new to the facility will need to produce evidence of being mycoplasma-free

This service takes 2-5 weeks to complete and includes the following:

• Thaw cells on mouse feeders or under feeder-free conditions

• Transition cells from feeder-dependent culture to feeder-free conditions if requested
• Expansion of cells
• Cryopreservation of expanded cells into 20 vials per cell line
• Mycoplasma testing
• Transfer of all frozen vials

Additional Notes:

• If thawing fails, PI will be notified immediately

This service establishes genome modified hESCs/hiPSCs using CRISPR/Cas9 technology

Prerequisites:

• Selection of a hESC or hiPSC line to be gene edited
• A Service Agreement must be executed for non-Penn users
• MTA must be executed if transferring a cell line to the Core
• All cell lines not generated by the Core must be shown evidence of being mycoplasma- free

This service takes 6-9 months to complete and includes the following:

• Consultation with the user to determine targeting strategy
• Design and construction of CRISPR gRNA(s) for targeting
• Validation of gRNA(s) in the targeting cell line
• Targeting of the cell line using validated gRNA(s)
• Screening of targeted clones
• Establishment of at least 2 correctly targeted clones
• Mycoplasma testing of the clones
• qPCR-based analysis for common karyotypic abnormalities of the clones
• Delivery of cryopreserved cells and written report

This Service provides the design and the construction (cloning) of a plasmid template used for targeting iPSCs using homology directed repair (e.g. template for generating a gene reporter)

Prerequisites:

• Selection of a hESC or hiPSC line to be gene edited
• A Service Agreement must be executed for non-Penn users
• MTA must be executed if transferring a cell line to the Core
• All cell lines not generated by the Core must be shown evidence of being mycoplasma- free

This service takes 3-5 weeks to complete and includes the following:

• Consultation with the user to determine targeting strategy
• Design and construction of the donor vector
• Delivery of the donor vector as a DNA maxiprep

This service establishes genome modified hESCs/hiPSCs using CRISPR/Cas9 technology

Prerequisites:

• Selection of a hESC or hiPSC line to be gene edited
• A Service Agreement must be executed for non-Penn users
• MTA must be executed if transferring a cell line to the Core
• All cell lines not generated by the Core must be shown evidence of being mycoplasma- free

This service takes 6-9 months to complete and includes the following:

• Consultation with the user to determine targeting strategy
• Design and construction of CRISPR gRNA(s) for targeting
• Validation of gRNA(s) in the targeting cell line
• Targeting of the cell line using validated gRNA(s) and HDR DNA donor template
• Screening of targeted clones
• Establishment of at least 2 correctly targeted clones
• Mycoplasma testing of the clones
• qPCR-based analysis for common karyotypic abnormalities of the clones
• Delivery of cryopreserved cells and written report

This service establishes genome modified hESCs/hiPSCs using CRISPR/Cas9 technology

Prerequisites:

• Selection of a hESC or hiPSC line to be gene edited
• A Service Agreement must be executed for non-Penn users
• MTA must be executed if transferring a cell line to the Core
• All cell lines not generated by the Core must be shown evidence of being mycoplasma-free

This service takes 6-9 months to complete and includes the following:

• Consultation with the user to determine targeting strategy
• Design and construction of CRISPR gRNA(s) for targeting
• Validation of gRNA(s) in the targeting cell line
• Design and synthesis of the ssODN donor template
• Targeting of the cell line using validated gRNA(s) and ssODN donor template
• Screening of targeted clones
• Establishment of at least 2 correctly targeted clones
• Mycoplasma testing of the clones
• qPCR-based analysis for common karyotypic abnormalities of the clones
• Delivery of cryopreserved cells and written report

This service provides iPSC-derived cardiomyocytes in monolayer cultures.  iPSCs are differentiated using cytokines and small molecules in defined media.  Cultures are then metabolically selected to enrich for cardiomyocytes and re-plated. These cells resemble immature cardiomyocytes and but also display a number of properties of adult cardiomyocytes.  

Prerequisites:

• Designation of iPSC line to be differentiated

This service takes 5-8 weeks to complete and includes the following: 

• Thaw cells and expand for differentiation
• Directed differentiation in monolayer cultures using SOP established by the Core
• Transfer of live monolayer cultures of beating cells in two 10-cm plates
• Approximate yield: 12-20 million cells

 Additional Notes:

• Additional plates can be purchased at a reduced price

This service can only be added in addition to iPSC-Derived Cardiomyocytes

This service provides iPSC-derived cardiomyocytes in monolayer cultures.  iPSCs are differentiated using cytokines and small molecules in defined media.  Cultures are then metabolically selected to enrich for cardiomyocytes and re-plated. These cells resemble immature cardiomyocytes and but also display a number of properties of adult cardiomyocytes. 

Prerequisites:

• Designation of iPSC line to be differentiated
• New iPSC lines will need to show evidence of being mycoplasma-free or this will be performed by the Core at extra charge (see Mycoplasma Testing service)

This service takes 5-8 weeks to complete and includes the following:

• Thaw cells and expand for differentiation
• Up to two rounds of pilot studies to determine optimal seeding density and conditions for differentiation 
• Directed differentiation in monolayer cultures
• Transfer of live monolayer cultures of beating cells in two 10-cm plates
• Approximate yield: 12-20 million cells

Additional Notes:

• Additional plates can be purchased at a reduced price.
• If differentiation fails in both pilot studies, PI will be notified immediately and the project will be discontinued.  Service fee still applies.
• More pilot studies can be performed using a different CM differentiation protocol after submitting a new request.

This service provides iPSC-derived endothelial cell in monolayer cultures.  iPSCs undergo directed differentiation using cytokines and small molecules to endothelial progenitors.   Cultures are then purified with endothelial specific cell surface markers to enrich for purity and further passaged.  These cells express CD31(PECAM) & CD144 (VE-cadherin) surface markers, uptake Ac-LDL, and form vessels when cultured with stromal cells in 3D cultures.

Prerequisites:

• Designation of iPS line to be differentiated
• New iPSC lines will need to show evidence of being mycoplasma-free

This service takes 4-6 weeks to complete and includes the following:

• Thaw cells and expand for differentiation
• Directed differentiation in monolayer cultures
• Magnetic activated cell sorting against endothelial markers
• Passaged in endothelial growth media for cell expansion
• Flow cytometry analysis of endothelial markers CD31/CD144
• Passage 3 live culture supplied in at least 2 6-well plates
• Approximate yield: 6 million cells
• Guaranteed purity > 90% CD31⁺/CD144⁺

Additional Notes:

• If differentiation fails, PI will be notified immediately
• Additional plates can be purchased at a reduced price

Under construction.  Please contact the Core director.

Under construction.  Please contact the Core director.

Prerequisites:

• Students must have at least one year of hands-on experience in mammalian cell culture

This is a hands-on training course in basic techniques in hESC/iPSC culture including cell thawing, expansion, identification by morphology, manual and enzymatic passaging, freezing, and feeder-free culture.  This course will take 2 weeks to complete (1-2 hours per day)

We provide limited tissue culture space to UPenn investigators who elect to perform their experiments in the iPSC core with guidance from the Core staff.   Investigators have full access to our hESC/iPSC dedicated mycoplasma-free cell culture rooms, consumables and some commonly used stem cell culture reagents, such as PBS and DMEM/F12.   All other reagents, such as stem cell medium, coating matrix are charged separately or provided by the user.

Prerequisites:

• Must have completed the hESC/hiPSC culture hands-on training course
• Must have approval from Core director following a consultation meeting

The Core has a number of wild-type control iPSC lines available to users.   Please inquire with the Core director at wenliyan@pennmedicine.upenn.edu.

R&D Systems cat no. 3433-005-01

Current Lot Protein Concentration: 8.32 mg/mL

Stem Cell Technologies Catalog # 100-0276

Miltenyi Biotec Catolog #130-104-368

Gibco Catalog # 10828028

Gibco Cat#17504-044

Gibco Cat#A1895601

100ug/mL stock, R&D Systems Cat# 338-AC-01M/CF

100ug/mL stock, R&D Systems Cat#233-FB-001MG/CF

100ug/mL stock, PeproTech Cat#AF-120-05ET