Identification of grafted bone marrow cells in the recipient tissues

Konference: 2011 7. Sympozium a workshop molekulární patologie a histo-cyto-chemie

Kategorie: Onkologická diagnostika

Téma: Postery

Číslo abstraktu: 012p

Autoři: Prof.MUDr. Jaroslav Mokrý, Ph.D.; prof. MUDr. Stanislav Filip, Ph.D., DSc.; Ing. Jaroslava Vávrová, Ph.D.; D. Čížková; pplk. doc. MVDr. Zuzana Šinkorová, CSc.; Doc.MUDr. Stanislav Mičuda, Ph.D.

Introduction: Identification of transplanted cells is facilitated when donor cells were tagged with endogenous vectors.

Aim: Comparison of efficacy of two different models based on bone marrow donor cells derived from transgenic ROSA26 and eGFP mice.

Material and methods: Full bone marrow or lin-CD117+ of B6;129S-Gt(ROSA)26Sor mice or C57BL/6-Tg(CAG-EGFP)C14Y01FM131Osb mice were transplanted into B6129SF2/J or C57BL/6J recipients subjected to 9Gy whole body lethal irradiation.

At different time points post transplantation (30, 60 and 180 days), cell settlement in recipient tissues including spleen, thymus, small intestine and liver was assessed histologically and verified by qPCR analysis.

Results: By day 30 transplanted cells infiltrated stromal components of examined organs as connective tissue wandering cells. In the thymus and spleen, grafted bone marrow-derived cells participated in lymphatic infiltration of both organs including thymic cortex and splenic nodules, indicating generation of donor-derived T- and B-lymphocytes. Transplanted animals were allowed to survive up to 6 months, which demonstrated establishment of a stable cellular chimerism. Data from qPCR were in a correlation with histological analysis.

Conclusion: eGFP mice represent a more reliable system over ROSA26 mice since GFP+ cells may be directly visualized in tissues (do not suffer from drawbacks associated with a poor penetration of X-Gal substrate and non-specifity of histochemical staining)and may be also directly analysed for flow cytometry. Histological examination of tissues obtained from transplanted mice is prior to PCR analysis because it allows not only to confirm identity, phenotype, status of transplanted cells and engraftment efficacy but also to specify their number, fate and distribution in recipient tissues.

This work was supported by MSM 0021620820.

Datum přednesení příspěvku: 29. 4. 2011