Cell selection by selective matrix adhesion is not sufficiently efficient for complete malignant cell depletion from contaminated human testicular cell suspensions.
OBJECTIVE
To investigate whether a selective matrix adhesion-based protocol can enrich germ cells and deplete cancer cells from contaminated human testicular cell suspensions.


METHODS
Artificially contaminated and control testicular cell suspensions were selectively cultured. After each experimental step, the presence of B cells was evaluated by fluorescence-activated cell sorting (FACS) and/or polymerase chain reaction (PCR).


METHODS
Academic medical center.


METHODS
Five vasectomy reversal patients willing to donate testicular tissue for research after informed consent.


METHODS
In vitro culture, magnetic-activated cell sorting (MACS), matrix adhesion, FACS, and PCR.


METHODS
FACS analysis for CD49f and human leukocyte antigen (HLA) class I expression; PCR analysis for a 120-bp fragment of the B-cell receptor.


RESULTS
With the use of a matrix adhesion-based protocol, CD49f(+) HLA class I(-) cells could be highly enriched, but HLA class I(+) CD49f(-) cells could still be detected. PCR proved that the cell suspensions contained malignant cells originating from the cell line used to contaminate them.


CONCLUSIONS
Even though an efficient enrichment of germ cells was achieved, the resulting population was not pure. Malignant cells were detected in selected cell suspensions of all five patients, demonstrating that the efficacy of this protocol is insufficient for clinical application.
Biology of the Testis Research Group, Department of Embryology and Genetics, Medical School and Universitair Ziekenhuis Brussel, Vrije Universiteit Brussel, Brussels, Belgium. mieke.geens@uzbrussel.be
Mieke
M
Geens
Ellen
E
Goossens
Herman
H
Tournaye
D000465:Algorithms; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D005109:Extracellular Matrix / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods; D005854:Germ Cells / Q000378:metabolism / Q000473:pathology / Q000502:physiology; D006801:Humans; D008297:Male; D013379:Substrate Specificity; D013535:Suspensions; D013736:Testicular Neoplasms / Q000473:pathology*; D013737:Testis / Q000737:chemistry / Q000473:pathology*
D013535:Suspensions