title: Isolation of germ cells from leukemia and lymphoma cells in a human in vitro model: potential clinical application for restoring human fertility after anticancer therapy.
abstract: More than 70% of patients survive childhood cancer, but chemotherapy and radiation therapy may cause irreversible impairment of spermatogenesis. To treat infertility secondary to anticancer treatment for childhood cancer, we have developed a procedure to isolate germ cells from leukemic mice by fluorescence-activated cell sorting with two surface markers, and transplantation of isolated germ cells successfully restored fertility without inducing leukemia. In the present study, we analyzed human germ cells and human malignant cells, including five leukemia cell lines and three lymphoma cell lines, by fluorescence-activated cell sorting with antibodies against MHC class I and CD45. Testicular specimens were obtained from a patient who underwent surgery for testicular rupture. In the high forward scatter and low side scatter region, no malignant cells were found in the MHC class I-negative and CD45-negative fraction (the germ cell fraction), with the exception of K562 cells. A total of 39.2% of the germ cells were found in the germ cell fraction. A total of 1.45% of K562 cells were found in the germ cell fraction. Treatment with IFNgamma induced the expression of MHC class I on K562 cells but not on germ cells and made it possible to isolate germ cells from K562 cells. In conclusion, we isolated human germ cells from malignant cells with two surface markers after treatment with IFNgamma. Immunophenotyping for each patient will be necessary before isolation and induction of surface marker will be clinically applicable.
authors.affiliation: Department of Urology, Osaka University Graduate School of Medicine, Suita Osaka, Japan.
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Fujita
authors.forename: Akira
authors.initials: A
authors.lastname: Tsujimura
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Miyagawa
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kiuchi
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Matsuoka
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Takao
authors.forename: Shingo
authors.initials: S
authors.lastname: Takada
authors.forename: Norio
authors.initials: N
authors.lastname: Nonomura
authors.forename: Akihiko
authors.initials: A
authors.lastname: Okuyama
mesh_terms: D045744:Cell Line, Tumor; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D064420:Drug-Related Side Effects and Adverse Reactions; D005298:Fertility / Q000187:drug effects / Q000528:radiation effects; D005434:Flow Cytometry / Q000379:methods*; D005854:Germ Cells / Q000166:cytology* / Q000378:metabolism; D018922:HL-60 Cells; D015234:HLA-A Antigens / Q000235:genetics / Q000378:metabolism; D015235:HLA-B Antigens / Q000235:genetics / Q000378:metabolism; D015236:HLA-C Antigens / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007246:Infertility / Q000209:etiology / Q000628:therapy; D019169:Jurkat Cells; D020014:K562 Cells; D007938:Leukemia / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D017493:Leukocyte Common Antigens / Q000235:genetics / Q000378:metabolism; D008223:Lymphoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008297:Male; D011878:Radiotherapy / Q000009:adverse effects; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013737:Testis / Q000166:cytology / Q000378:metabolism; D020298:U937 Cells
chemical_list: D015234:HLA-A Antigens; D015235:HLA-B Antigens; D015236:HLA-C Antigens; D017493:Leukocyte Common Antigens