Figure 2

Immunological survival of hMSC-EGFP cell transplants in rat spinal cord. (A) Histological assessment of hMSC-EGFP cell transplant survival in rat spinal cord without systemic immune suppression at five time points post transplantation (day 1 and week 1–4). First row: hematoxylin-eosin (HE) staining indicating localisation and general appearance of transplantation site. Second row: direct EGFP fluorescence indicating the presence or absence of EGFP positive hMSC-EGFP cell transplants. Third row: immuno-histochemical staining for EGFP indicating the presence or absence of EGFP positive hMSC-EGFP cell transplants. Fourth row: immuno-histochemical staining for CD68 indicating macrophage infiltration into the transplantation site. All slides were examined using a conventional light/fluorescence microscope and digital pictures were taken under magnification as indicated. Representative pictures were chosen from multiple hMSC-EGFP cell transplanted spinal cords analysed for day 1 (n = 2), week 1 (n = 2), week 2 (n = 2), week 3 (n = 2), and week 4 (n = 6) post-transplantation. (B) Molecular and histological assessment of hMSC-EGFP cell transplant survival in rat spinal cord under systemic immune suppression (subcutaneous 10 mg/kg/day cyclosporin A) at four time points post transplantation (day 1 and week 1–3). DAY 1: Real-time RT-PCR analysis on mRNA isolated from hMSC-EGFP cell transplanted spinal cords on day 1 post-transplantation (n = 2) indicating the presence of EGFP mRNA transcripts in vivo in spinal cord following hMSC-EGFP cell transplantation. WEEK 1: direct EGFP fluorescence and immuno-histochemical staining for EGFP indicating the presence of EGFP positive hMSC-EGFP cell transplants on week 1 post-transplantation. Representative pictures were chosen from multiple hMSC-EGFP cell transplanted spinal cords analysed 1 week post-transplantation (n = 5). WEEK 2: HE staining, direct EGFP fluorescence and immuno-histochemical staining for EGFP indicating the presence of EGFP positive hMSC-EGFP cell transplants on week 2 post-transplantation. Representative pictures were chosen from multiple hMSC-EGFP cell transplanted spinal cords analysed 2 weeks post-transplantation (n = 3). WEEK 3: Standard PCR and RT-PCR analysis on DNA and mRNA isolated from hMSC-EGFP cell-transplanted spinal cords 3 weeks post-transplantation indicating the presence of EGFP DNA sequences and EGFP mRNA transcripts in vivo in spinal cord following hMSC-EGFP cell transplantation.