The Stem Cell Engineering facility at CRTD (formerly known as ES/iPS Facility) provides the following services:

1. Generation and characterization of induced pluripotent stem cells

The facility generates intergation/transgene free human induced pluripotent stem (hiPS) cells from diverse kinds of somatic cells including, but not limited to, cells from cord and peripheral blood, various fibroblasts, brain endothelial cells.

The generated iPSCs are characterized for pluripotency using the following markers: 

• FLOW analysis of pluripotency markers

• Quantitative real time PCR analysis for pluripotency markers

• Quantitative real time PCR analysis for spontaneous and/or cytokine/growth factor assisted in vitro three germ layer differentiations

• Short Tandem Repeat (STR) analysis

• Karyotyping

• Immunochemistry of pluripotency markers

• Immunochemistry of three germ layer markers

• Mycoplasma testing

• Post Thaw viability

The customer of the facility chooses the number/level of characterization.

2. Engineering of hES/hiPS cell lines

This service includes the following:

• Generation of isogenic hES/hiPS cell lines
   
• Gene knock in/out in hES/hiPS cell lines
   
• Gene/tissue specific/double hES/hiPS cell reporters (fluorescent and/or drug resistant)
   
• Seamless gene corrections of point or short mutations in ES/iPS cell lines

3. Generation of cortical neural precursor cells (NPCs) from hES/hiPS cells

The facility helps optimize protocols and generates NPCs from hES/hiPS cells and provides them as frozen stocks to investigators.
These NPCs can be plated in neuronal differentiation media giving rise to cortical neurons of upper and lower cortical layers.

4. Culture and adoption of hES/hiPS cells

• The facility can develop feeder independent cultures of hES/hiPS cells from feeder dependent cultures to substrates like Matrigel, Vitronectin, Geltrex and Synthamax.

  The adaption will include not only substrate adaption but also changing/adapting of media for support and maintenance of these lines on the adapted substrates.

• hES/hiPS cell lines can be made to adapt to single cell passaging (TrypLe) on feeder or feeder independent cultures on Matrigel, Vitronectin, Geltrax and Sythamax substrates.