Materials and Reagents

1. Human or mouse mesenchymal stem cells (cultured, treated or transfected)
   Note: If using the primary bone marrow stem cells (attached cells from bone marrow culturing), it’s better to use cells in low passages (not over passage 3); if using MSC cell lines, it’s also better to use low passages than high passages.
   
2. Mice [NOD.CB17-Prkdc/J mice (NOD/SCID), around 8 weeks old]
   
3. Hydroxyapatite/tricalcium phosphate (HA/TCP) (Zimmer, catalog number: 60-130)
   
4. 4% paraformaldehyde
   
5. Hematoxylin/Eosin (H&E) staining solution
   
6. Formic acid solution (see Recipes)
   

Equipment

1. 1 ml syringes
   
2. Cotton sticks (Pro-Ophtha)
   
3. Scalpels
   
4. Racks (for fixing the syringes in vertical position in cell incubator)
   
5. Tools for mouse surgery (scalpel, forceps, needle holder, scissors, sutures, sterile swabs, sterile drapes)
   
6. Scan microscopy (Leica Microsystems or any scan microscopy)
   
7. Computer (with image analysis software, such as ImageJ^® or Photoshop CS^®)
   

Software

1. ImageJ^® or Photoshop CS^®

Procedure