Stem Cell Core

The Stem Cell Core houses 3 cell culture suites, 2 lab benches, and cell analysis. Each cell culture suite is outfitted with 3-4 bio-safety cabinets, 6-8 CO2 incubators, a low oxygen incubator, and inverted microscopes and requisite equipment for stem cell culture. The STEM Core is located adjacent to the Flow Cytometry Core Facility enabling efficient cell sorting and cytometry analysis experiments.

Instrumentation

Cell Imaging
2x Dissection microscopes for manual isolation and manipulation of stem cell colonies
SeaHorse 96XFe for cell analysis

Cell Culture
11x Biological safety cabinets
18x C02 Incubators
3x N2/C02 Incubators (hypoxic conditions)
3x Bench top centrifuges
3x 37°C Water baths

Transfection
LifeTech Neon – Tip based electroporation (adjustable parameters)
MaxCyte ExPERT A+X electroporator (Cuvette-based; fixed parameter)

Centrifugation
Optima L-80XP Ultracentrifuge
Allegra X-15R Plate centrifuge
Bench top temperature controlled centrifuge & microcentrifuge

Refrigerators and Freezers
3x -80°C freezers
10x -20°C freezers
6x 4°C refrigerators
2x High capacity deli refrigerators
2x MVE 800 Series Cyrostorage freezers – 11664 vial capacity, auto fill controllers

Services

Biopsy processing (fibroblast derevation)
Reprogramming from fibs or PBMCs
Custom genome editing
iPSC QC (by request)

• Karyotyping
• Mycoplasma testing
• STR
• Undiff
• TriLin
• Sev retention

Fib QC (by request)

• Mycoplasma
• Growth rate
• STR profiling (human only)

Genome editing QC (by request)

• Myco plasma
• Allele sequencing
• STR

Access:
The Stem Cell Core Facility is available to researchers 7 days a week, 24 hours a day. Access to the facility must be arranged in advance and approved by the STEM Core.

Conditions of Use:

1. The Core Director must first approve all new cell lines that will be brought into the facility, including both live and frozen cell cultures, from both commercial and academic sources. All new, approved cell lines will be cultured under quarantine conditions until they have tested clean for mycoplasma in 2 consecutive tests.
2. All new STEM Core users must be approved by, and go through a brief orientation with the Core Director, or Lab Manager.
3. Do not bring previously opened reagents, supplies or materials from other labs into the facility. In some instances this may be necessary, but must first be cleared with the Core Director. Unopened, direct from supplier, cell culture media and materials may be brought into the facility.

SCRO approvals, ethics training, and MTAs:
Any research involving human pluripotent stem cells at the Salk Institute will require:

1. an SCRO approved protocol
and
2. ethics training for the researchers and PI involved with the experiments.

Please consult the SCRO Website for information on fulfilling these requirements. The requirements for approval will vary depending on the research project. The Core Director and staff can assist new researchers with the process of filling SCRO protocols. For training purposes, new investigators can begin working under the existing stem cell culture techniques training protocol. However, as the investigator(s) develop their own research project, separate SCRO approval must be obtained.

Information on ethics training can be found online here»

Salk researchers working in the STEM Core are covered under the existing MTAs for the cell lines provided. Should an investigator want to culture one of the cell lines below in their own lab, a separate MTA would need to be executed.

Cell Lines Available

All cell lines are karyotypically stable, pathogen free, and cultured without antibiotics. Additional cell lines available upon request.

Human Embryonic Stem Cell Lines Available:

Cell Line	Passage # on MEFS	Passage # on mtg/TeSR
H1	p32	p39
H7	p26	p28
H9	p33	p35
H13	p30	p32
HUES-3	p29	p30
HUES-6	p21	p24
HUES-8	p32	–
HUES-9	–	p33
LSJ-1	–	p24

Human Induced Pluripotent Stem Cell Lines Available:
Disease Specific iPSCs: Are you looking for a specific disease-linked iPSC line? It might already exist and we might be able to help you find it.

Cell Line	Passage # on MEFS	Passage # on mtg/TeSR
BJ-iPSC	p10-28	p10-12
WiCell-iPSC	–	p33

Other cell types available:
Keratinocytes-Adult
Keratinocytes-Neo Natal
BJ Fibroblasts
IMR90s
OP9s
293Ts
HFFs
MEFs
HUVEC

MTA Considerations
Salk researchers working in the STEM Core are covered under the existing MTAs for the cell lines provided. Should an investigator want to culture one of these cell lines in their own lab, a separate MTA would need to be executed.