Staff
Zack Wang, PhD
Core Director
207-885-8313
wangz@mmc.org

Melanie Arzigian, BS
Core Manager
207-885-8314
arzigm@mmc.org

Overview:
The cell separation branch of this core provides instrumentation and expertise for cell sorting, using FACS and MACS technologies. The ES cell branch of this core provides an institutional resource for the study of embryonic stem (ES) cells, having a dedicated ES cell culture facility equipped with tissue culture microscopes and a fluorescence microscope, tissue-culture hoods, CO2 incubators, LN2 storage, and ES cell specific reagents.

For cell separation interests, this core provides two instrumentation resources: the FACSAria flow cytometer from BD Biosciences and the autoMACS separator from Miltenyi Biotec.  The FACSAria is the first fixed alignment benchtop high-speed sorter. With its digital electronics and various pressures, it can process 70,000 cells per second. Other features include its fixed-optics, multi-well sorting and the capacity of analyzing 15 parameters simultaneously.
The autoMACS from Miltenyi Biotec uses MAgnetic Cell Separation technology. Magnetic Cell Separation utilizes highly specific isolation of magnetically labeled cells. The cells of interest are labeled, passed through a separation column and are separated from the unlabeled cells. The system is fast, efficient and completely compatible with the flow cytometers.

For ES cell interests, this core provides technical support, training, and services for stem cell research.  ES cell lines will be maintained with appropriate quality assurance in the facility.  The technical support provided by the facility will include teaching of methods for the maintenance, transduction, and differentiation of the cells.  The facility will provide short-term culture and manipulation of the cells for individual investigators.  In addition, the core will provide ready access of ES cell reagents (RNAs, Genomic DNAs, and proteins) to individual investigators without either the need for specialized space, or the need to develop specialized technical skills required for ES cell work.  The ES cell core will also provide technical support and training for human ES cell culture (NIH approved human ES cell lines: H1 and H9).  A variety of mouse embryonic feeder (MEF) cells, inactivated and plated, is also available through this core.

 
Services Available:

The Core facility provides the following:

Cell Separation:

• Service contracted core instruments, and the instrument reagents such as sheath fluid and running buffers
• Technician performed FACSAria cell sorting upon request
• Individual use of the autoMACS
• FACSAria overview and autoMACS training for interested individuals
• Technical assistance in cell preparation for cell sorting

ES cell:

• Plated, mitomycin C (or irradiation) inactivated feeder cells (mouse embryonic fibroblasts: MEF’s)
• Murine ES cells for in vitro and in vivo study
• RNA and protein from undifferentiated murine or human ES cells, or from differentiated murine or human embryonic bodies (EB’s)
• Quality reagents for ES cell culture, including mediums containing FBS and LIF.
• Freezing, thawing, and expansion of ES cell lines
• Murine and human ES cell culture counseling

Service Requests:
A Service Request Form for MMCRI Users  and a Service Request Form for non-MMCRI Users can be downloaded from the website.  The form must be filled out completely with as much detailed information as possible.  The completed form must include the signature and account number of the Principal Investigator of the lab requesting services.  Completed forms should be submitted to Melanie Arzigian.