Long-term follow-up of porcine male germ cells transplanted into mouse testes

Zygote ◽  
2007 ◽  
Vol 15 (4) ◽  
pp. 325-335 ◽  
Author(s):  
Y-J. Choi ◽  
J-K. Park ◽  
M-S. Lee ◽  
J.D. Ahn ◽  
K-C. Hwang ◽  
...  
Keyword(s):  
Germ Cells ◽  
Detection System ◽  
Phylogenetic Distance ◽  
Male Germ Cells ◽  
Reliable Technique ◽  
Donor Cells ◽  
Male Donor ◽  
Long Term Follow Up ◽  
Foreign Donor

SummaryThis study investigated the effect of increased phylogenetic distance on the outcome of spermatogonial transplantation, with porcine donors and mice recipients. It was designed to develop a technique for detecting foreign donor cells in recipient animals. Porcine male germ cells were harvested from postnatal male testes and incubated with the lipophilic membrane dye PKH-26. For transplantation, approximately 106 PKH-26-labelled porcine male germ cells were injected into the efferent ducts of mouse testes. Animals were sacrificed at post-graft days 1, 10, 30, 45, 60 and 150 (n = 5 each). Serial frozen sections of explanted testes were prepared for detecting labelled cells. Transplanted porcine donor cells were easily detected in the recipient tubules for 8 weeks. After transplantation, we could detect both incorporation into the basement membrane and differentiation of grafted porcine donor cells by our double detection system, using PKH staining and slide PCR. However, our RT-PCR and apoptosis results revealed that most of the grafted porcine male donor cells could not differentiate past early-meiotic spermatocytes. We could induce partial differentiation of xenografted porcine donor cells in mouse testes, but not full induction of spermatogenesis. We have developed a very reliable technique for detecting foreign donor cells in recipient animals using a combination of PKH staining and slide PCR methods. Our results provide a valuable experimental model for applying and evaluating this technology in other species.

A Potential Alternative Strategy for Myoblast Transfer Therapy: The use of Sliced Muscle Grafts

1996 ◽  
Vol 5 (3) ◽  
pp. 421-429 ◽  
Author(s):  
Ying Fan ◽  
Manfred W. Beilharz ◽  
Miranda D. Grounds
Keyword(s):  
Specific Probe ◽  
Poor Survival ◽  
Term Survival ◽  
Alternative Approach ◽  
Donor Cells ◽  
Male Donor ◽  
Potential Alternative ◽  
Female Host ◽  
Myoblast Transfer

Excellent long-term survival (up to 1 yr) of donor skeletal muscle cells was demonstrated using a mouse Y-chromosome specific probe, following the transplantation of grafts of whole muscles from male “normal” C57B1/10Sn mice into dystrophic muscles of female host mice. After the transplantation of equivalent sliced muscle grafts there was extensive movement of the male donor cells and fusion with host myofibres. This contrasts with the extremely poor survival of isolated myoblasts after injection into the same mouse model for Duchenne muscular dystrophy. The use of sliced muscle grafts may therefore represent a potential alternative approach to myoblast transfer therapy.

Survival of Donor Leukocyte Subpopulations in Immunocompetent Transfusion Recipients: Frequent Long-Term Microchimerism in Severe Trauma Patients

Blood ◽  
1999 ◽  
Vol 93 (9) ◽  
pp. 3127-3139 ◽  
Author(s):  
Tzong-Hae Lee ◽  
Teresa Paglieroni ◽  
Hitoshi Ohto ◽  
Paul V. Holland ◽  
Michael P. Busch
Keyword(s):  
Magnetic Beads ◽  
Elective Surgery ◽  
Quantitative Polymerase Chain Reaction ◽  
Severe Trauma ◽  
Hla Typing ◽  
Trauma Patients ◽  
Donor Cells ◽  
Male Donor ◽  
Mutual Tolerance

We recently reported detection of a transient increase in circulating donor leukocytes (WBCs) in immunocompetent recipients 3 to 5 days posttransfusion (tx) (Blood 85:1207, 1995). We have now characterized survival kinetics of specific donor WBC subsets in additional tx populations. Eight female elective surgery patients (pts) were sampled pre-tx and on days 1, 3, 5, 7, and 14 post-tx. Ten female trauma pts transfused with a total of 4 to 18 U of relatively fresh red blood cells were sampled up to 1.5 years post-tx. WBC subsets from frozen whole blood were isolated using CD4, CD8 (T cell), CD15 (myeloid), and CD19 (B cell) antibody-coated magnetic beads. Donor WBCs were counted by quantitative polymerase chain reaction (PCR) of male-specific sex determining region (SRY) sequences. PCR HLA typing and mixed leukocyte reaction (MLR) between recipient and donor WBCs were performed on two of the trauma tx recipients who had long-term chimerism of donor cells post-tx. In 6 of 8 female surgery pts, circulating CD4+ male donor cells peaked at day 3 or 5 (0.01 to 1 cell/μL), followed by clearance by day 14. In 7 of 10 female trauma pts, we observed multilineage persistence of male donor WBCs (CD4, CD8, CD15, CD19) for 6 months to 1.5 years post-tx at concentrations of 10 to 100 cells/μL. In 2 trauma recipients studied, MLR showed no, or very low, response to WBC of the single donor implicated as the source of microchimerism by HLA typing. Establishment of long-term multilineage chimerism in trauma recipients is probably caused by engraftment of donor stem cells and mutual tolerance between recipient and donor leukocytes. A better understanding of factors determining clearance versus chimerism of transfused leukocytes is critical to prevention of alloimmunization and transfusion-induced graft-versus-host disease, and, potentially, to induction of tolerance for transplantation.

Survival of donor cells 25 years after intrauterine transfusion

Blood ◽  
2000 ◽  
Vol 95 (8) ◽  
pp. 2709-2714
Author(s):  
Henk E. Viëtor ◽  
Eric Hallensleben ◽  
Simone P. M. J. van Bree ◽  
Ellen M. W. van der Meer ◽  
Suzanne E. J. Kaal ◽  
...  
Keyword(s):  
Y Chromosome ◽  
T Lymphocyte ◽  
Cytotoxic T Lymphocyte ◽  
Good Health ◽  
Fish Analysis ◽  
Chronic Antigenic Stimulation ◽  
Donor Cells ◽  
Intrauterine Transfusion ◽  
Male Donor

Persistence of donor leukocytes in the circulation of recipients of intrauterine transfusion (IUT) has been observed up to 5 years after birth. The aim of this study was to determine whether transfusions with nonirradiated, nonleukocyte-depleted donor blood during the fetal period resulted in long-term immunomodulation of the recipient. Twenty-four surviving IUT recipients between 1966 and 1976 were tested for autoimmune disease and autoantibodies at follow-up. Ten had sex-mismatched donors and were therefore informative for chimerism studies using fluorescence in situ hybridization (FISH). Seven female recipients could be tested for chimerism using a Y- chromosome–specific polymerase chain reaction (PCR) because they received at least 1 IUT from a male donor. Nine recipients could be studied for cytotoxic T-lymphocyte precursor (CTLp) and helper T-lymphocyte precursor (HTLp) frequencies because the original donors were available for testing. All surviving IUT recipients were in good health at the time of the examination, and routine laboratory testing revealed no abnormalities. None of the IUT recipients were chimeric as determined by FISH analysis, but Y-chromosome–specific sequences were detected by PCR in 6 of the 7 women. However, the CTLp and HTLp frequencies of the IUT recipients against the donors were comparable to those of the controls. The current study provides evidence that IUT can result in the persistence of donor cells in the recipient for a period longer than 20 years but that it is not associated with immunotolerance or with signs of chronic antigenic stimulation.

JMJD1C, a JmjC Domain-Containing Protein, Is Required for Long-Term Maintenance of Male Germ Cells in Mice1

2013 ◽  
Vol 89 (4) ◽  
Author(s):  
Shunsuke Kuroki ◽  
Mika Akiyoshi ◽  
Mikiyo Tokura ◽  
Hitoshi Miyachi ◽  
Yuji Nakai ◽  
...  
Keyword(s):  
Germ Cells ◽  
Male Germ Cells ◽  
Jmjc Domain ◽  
Term Maintenance

Survival of Donor Leukocyte Subpopulations in Immunocompetent Transfusion Recipients: Frequent Long-Term Microchimerism in Severe Trauma Patients

Blood ◽  
1999 ◽  
Vol 93 (9) ◽  
pp. 3127-3139 ◽  
Author(s):  
Tzong-Hae Lee ◽  
Teresa Paglieroni ◽  
Hitoshi Ohto ◽  
Paul V. Holland ◽  
Michael P. Busch
Keyword(s):  
Magnetic Beads ◽  
Elective Surgery ◽  
Quantitative Polymerase Chain Reaction ◽  
Severe Trauma ◽  
Hla Typing ◽  
Trauma Patients ◽  
Donor Cells ◽  
Male Donor ◽  
Mutual Tolerance

Abstract We recently reported detection of a transient increase in circulating donor leukocytes (WBCs) in immunocompetent recipients 3 to 5 days posttransfusion (tx) (Blood 85:1207, 1995). We have now characterized survival kinetics of specific donor WBC subsets in additional tx populations. Eight female elective surgery patients (pts) were sampled pre-tx and on days 1, 3, 5, 7, and 14 post-tx. Ten female trauma pts transfused with a total of 4 to 18 U of relatively fresh red blood cells were sampled up to 1.5 years post-tx. WBC subsets from frozen whole blood were isolated using CD4, CD8 (T cell), CD15 (myeloid), and CD19 (B cell) antibody-coated magnetic beads. Donor WBCs were counted by quantitative polymerase chain reaction (PCR) of male-specific sex determining region (SRY) sequences. PCR HLA typing and mixed leukocyte reaction (MLR) between recipient and donor WBCs were performed on two of the trauma tx recipients who had long-term chimerism of donor cells post-tx. In 6 of 8 female surgery pts, circulating CD4+ male donor cells peaked at day 3 or 5 (0.01 to 1 cell/μL), followed by clearance by day 14. In 7 of 10 female trauma pts, we observed multilineage persistence of male donor WBCs (CD4, CD8, CD15, CD19) for 6 months to 1.5 years post-tx at concentrations of 10 to 100 cells/μL. In 2 trauma recipients studied, MLR showed no, or very low, response to WBC of the single donor implicated as the source of microchimerism by HLA typing. Establishment of long-term multilineage chimerism in trauma recipients is probably caused by engraftment of donor stem cells and mutual tolerance between recipient and donor leukocytes. A better understanding of factors determining clearance versus chimerism of transfused leukocytes is critical to prevention of alloimmunization and transfusion-induced graft-versus-host disease, and, potentially, to induction of tolerance for transplantation.

Survival of donor cells 25 years after intrauterine transfusion

Blood ◽  
2000 ◽  
Vol 95 (8) ◽  
pp. 2709-2714 ◽  
Author(s):  
Henk E. Viëtor ◽  
Eric Hallensleben ◽  
Simone P. M. J. van Bree ◽  
Ellen M. W. van der Meer ◽  
Suzanne E. J. Kaal ◽  
...  
Keyword(s):  
Y Chromosome ◽  
T Lymphocyte ◽  
Cytotoxic T Lymphocyte ◽  
Good Health ◽  
Fish Analysis ◽  
Chronic Antigenic Stimulation ◽  
Donor Cells ◽  
Intrauterine Transfusion ◽  
Male Donor

Abstract Persistence of donor leukocytes in the circulation of recipients of intrauterine transfusion (IUT) has been observed up to 5 years after birth. The aim of this study was to determine whether transfusions with nonirradiated, nonleukocyte-depleted donor blood during the fetal period resulted in long-term immunomodulation of the recipient. Twenty-four surviving IUT recipients between 1966 and 1976 were tested for autoimmune disease and autoantibodies at follow-up. Ten had sex-mismatched donors and were therefore informative for chimerism studies using fluorescence in situ hybridization (FISH). Seven female recipients could be tested for chimerism using a Y- chromosome–specific polymerase chain reaction (PCR) because they received at least 1 IUT from a male donor. Nine recipients could be studied for cytotoxic T-lymphocyte precursor (CTLp) and helper T-lymphocyte precursor (HTLp) frequencies because the original donors were available for testing. All surviving IUT recipients were in good health at the time of the examination, and routine laboratory testing revealed no abnormalities. None of the IUT recipients were chimeric as determined by FISH analysis, but Y-chromosome–specific sequences were detected by PCR in 6 of the 7 women. However, the CTLp and HTLp frequencies of the IUT recipients against the donors were comparable to those of the controls. The current study provides evidence that IUT can result in the persistence of donor cells in the recipient for a period longer than 20 years but that it is not associated with immunotolerance or with signs of chronic antigenic stimulation.

scholarly journals Male germ cells support long-term propagation of Zika virus

2018 ◽  
Vol 9 (1) ◽  
Author(s):  
Christopher L. Robinson ◽  
Angie C. N. Chong ◽  
Alison W. Ashbrook ◽  
Ginnie Jeng ◽  
Julia Jin ◽  
...  
Keyword(s):  
Germ Cells ◽  
Zika Virus ◽  
Male Germ Cells

scholarly journals Usefulness of real-time PCR in long-term follow-up of follicular lymphoma patients.

2007 ◽  
Vol 54 (1) ◽  
pp. 135-142 ◽  
Author(s):  
Andrzej Tysarowski ◽  
Anna Fabisiewicz ◽  
Ewa Paszkiewicz-Kozik ◽  
Jadwiga Kulik ◽  
Jan Walewski ◽  
...  
Keyword(s):  
Follicular Lymphoma ◽  
Real Time ◽  
Real Time Pcr ◽  
Residual Disease ◽  
Fusion Gene ◽  
Detection System ◽  
Breakpoint Region ◽  
Long Term Follow Up

The aim of this study was to evaluate the usefulness of quantitative real-time PCR (RQ-PCR) for the monitoring of molecular remission in follicular lymphoma (FL) patients during long-term follow-up. RQ-PCR by the use of TaqMan detection system is a sensitive tool to monitor minimal residual disease (MRD) in FL through amplification of the t(14;18) fusion gene during and post-therapy. In most cases the breakpoint region occurs within the major breakpoint region (MBR). Among 75 patients diagnosed with FL, cells harboring the fusion gene BCL2/JH were found in peripheral blood of 31 patients (41%). We further monitored 30 of these patients in a period varying from 6 months to 5 years by RQ-PCR. In our study the level indicating the possibility of the presence of MRD was established at more than five t(14;18)-positive cells in the background of 83,000 normal cells. The results of this work also confirmed that the presence of MRD detected by RQ-PCR is an indication for careful observation of patients because of a higher risk of disease recurrence.

scholarly journals Are zygomatic implants a simple and reliable technique for the stabilization of obturator prostheses? Case report and review of the literature

2020 ◽  
Vol 26 (2) ◽  
pp. 12
Author(s):  
Benjamin Sender ◽  
Thibault Lacroix ◽  
Philippe Jaby ◽  
Anne-Gaelle Chaux-Bodard
Keyword(s):  
Reliable Technique ◽  
Surgical Closure ◽  
Implant Placement ◽  
Zygomatic Implants ◽  
Significant Difference ◽  
Oncologic Patients ◽  
Long Term Follow Up ◽  
Obturator Prosthesis

Zygomatic implants have been used for several years for the treatment of extremely resorbed maxilla. Indications were extended for oral rehabilitations after maxillectomy in oncologic patients. A 24-year old patient with a triple A syndrome who underwent a left maxillectomy due to a spinocellular tumour was addressed for prosthetic rehabilitation. As his obturator prosthesis failed, surgical closure of the defect combined with 2 zygomatic implants to support the prosthesis was proposed. Despite a small persistent oro-antral fistula, the new obturator prosthesis restored the patient's functions and esthetics and improved his quality of life. The literature reports less than 40 cases of maxillectomy patients rehabilitated with zygomatic implants (with or without flap closure of the defect). Regardless of implant placement, there is no significant difference between reconstructive surgery and obturator prosthesis. Thus, zygomatic implants seem to be a reliable method for the stabilization of obturator prosthesis, without complex surgical procedure. Nonetheless, reservations should be expressed given the lack of data in terms of long-term follow up.

scholarly journals Surgical extrusion: A reliable technique for saving compromised teeth. A 5-years follow-up case report

2018 ◽  
Vol 32 (1) ◽  
pp. 25-30
Author(s):  
Jenner Argueta ◽  
Ana Orellana ◽  
Gianluca Plotino
Keyword(s):  
Case Report ◽  
Root Resorption ◽  
Anterior Tooth ◽  
Current Case ◽  
Reliable Technique ◽  
Anterior Teeth ◽  
Long Term Follow Up ◽  
Crown Lengthening

Aim: To present a long term follow up clinical case in which a compromised anterior tooth was saved by a surgical extrusion procedure. Summary: Although different techniques have been suggested for clinical crown lengthening in the anterior zone, some of them have limitations in terms of aesthetics and procedural requirements. The current case report demonstrates how a simplified surgical extrusion procedure was successfully performed for saving a severely damaged anterior tooth; furthermore, it is possible to apply the technique described in this case using minimum and simple armamentarium like a scalpel, elevators, forceps and splinting flexible cord. Key-learning points: Saving severely compromised anterior teeth is possible by applying surgical extrusion techniques when crown-root ratio allows it. Risk of root resorption or ankylosis is minimum.

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