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Isolation of decidual lymphocytes from chorionic villus samples: phenotypic analysis and growth in vitro
M K Haynes1, M T Flanagan, B Perussia
1Department of Medicine, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Insights
Chorionic villus samples (CVS) contain decidual natural killer (NK) cells and T cells. These cells proliferate in response to interleukin-2 (IL-2) and can be cultured for further study.
Area of Science:
- Immunology
- Reproductive Biology
- Cell Biology
Background:
- Chorionic villus samples (CVS) contain granulated cells resembling large granular lymphocytes.
- Decidual lymphocytes play a crucial role in pregnancy maintenance.
Purpose of the Study:
- To determine if CVS can serve as a source of decidual lymphocytes.
- To characterize the phenotype and in vitro behavior of lymphocytes derived from CVS.
Main Methods:
- Phenotypic characterization using two-color immunofluorescence.
- In vitro culture of CVS-derived cells with recombinant interleukin-2 (rIL-2).
- Selective culture techniques involving co-culture with an irradiated B lymphoblastoid cell line.
Main Results:
- CVS-derived lymphocytes include a major fraction of decidual NK cells (CD56+bright, CD16-).
- T cell populations (CD4+ and CD8+) are present in CVS, with a reduced ratio compared to peripheral blood.
- Both T cells and CD56+bright cells proliferate in response to rIL-2; preferential growth of CD56+bright cells is achieved with selective culture.
Conclusions:
- CVS is a source of decidual NK cells and T cells that respond to rIL-2 and stimulator cells.
- Established culture techniques enable further investigation into decidual NK cell and T cell maturation and activation.
Problem:
Giemsa stained cell isolates prepared from chorionic villus samples (CVS) contain granulated cells morphologically similar to large granular lymphocytes.
Method:
Phenotypic characterization of these cellular isolates by two-color immunofluorescence and subsequent in vitro culture in the presence of recombinant interleukin-2 (rIL-2) were done in order to determine whether CVS could serve as a source of decidual lymphocytes.
Results:
A major fraction of the CVS-derived lymphocytes were characterized as decidual NK cells, exhibiting high levels of CD56 expression (CD56+bright), without concomitant expression of CD16. The T cell population present in CVS-derived lymphocytes contained both CD4+ and CD8+ cells in a ratio somewhat reduced compared to that found in peripheral blood. While both T cells and CD56+bright cells from CVS proliferate in vitro in response to rIL-2 alone, preferential growth of CD56+bright cells was accomplished using a selective culture technique wherein co-culture with an irradiated, B lymphoblastoid cell line promoted the growth of CD56+ cells.
Conclusion:
CVS contains decidual NK cells and T cells that proliferate in response to rIL-2 and/or third party stimulator cells. These culture techniques will allow investigations into the maturation and/or activation of decidual NK cells and T cells.