Cell Transplantation Techniques Research Articles

The effectiveness of rigid pericardial endoscopy for minimally invasive minor surgeries: cell transplantation, epicardial pacemaker lead implantation, and epicardial ablation

BackgroundThe efficacy and safety of rigid pericardial endoscopy as the promising minimally invasive approach to the pericardial space was evaluated. Techniques for cell transplantation, epicardial pacemaker lead implantation, and epicardial ablation were developed.MethodsTwo swine and 5 canines were studied to evaluate the safety and efficacy of rigid pericardial endoscopy. After a double pericardiocentesis, a transurethral rigid endoscope was inserted into the pericardial space. The technique to obtain a clear visual field was examined, and acute complications such as hemodynamic changes and the effects on intra-pericardial pressure were evaluated. Using custom-made needles, pacemaker leads, and forceps, the applications for cell transplantation, epicardial pacemaker lead implantation, and epicardial ablation were also evaluated.ResultsThe use of air, the detention of a stiff guide wire in the pericardial space, and the stretching of the pericardium with the rigid endoscope were all useful to obtain a clear visual field. A side-lying position also aided observation of the posterior side of the heart. As a cell transplantation methodology, we developed an ultrasonography-guided needle, which allows for the safe visualization of transplantation without major complications. Pacemaker leads were safely and properly implanted, which provides a better outcome for cardiac resynchronizing therapy. Furthermore, the success of clear visualization of the pulmonary veins enabled us to perform epicardial ablation.ConclusionsRigid pericardial endoscopy holds promise as a safe method for minimally invasive cell transplantation, epicardial pacemaker lead implantation, and epicardial ablation by allowing clear visualization of the pericardial space.

Modified technique for spermatogonial stem cell transplantation into the seminiferous tubules in mouse model

This study aimed to develop a modified technique for spermatogonial stem cell (SSC) transplantation with the aid of an operating microscope in an infertile mouse model. Male neonatal C57BL/6 (B6) mice served as SSC donors. SSCs labeled with the PKH26-GL marker were detected by flow cytometry to verify purity. Adult B6 males were rendered infertile by busulfan treatment as the recipient. One month later the SSC suspension was delivered into recipient seminiferous tubules by manual microinjection under microscope with 100x magnification. This was compared to the conventional mechanical micromanipulator method via efferent ducts, rete testis, and seminiferous tubules, respectively. The volume injected and time required in the different procedures were compared. The recipient accepted manual microinjection via seminiferous tubules was subjected to histology, confocal laser scanning microscopy, and real-time fluorescent PCR at different checkpoints after transplantation. Positive controls received neither busulfan treatment nor transplantation. Negative controls were injected with an equal amount of transplant medium. The results showed that manual microinjection took 10 minutes per testis for the complete delivery of 50 µl of the SSC suspension, which was significantly less time-consuming and delivered a larger volume of SSC suspension than other methods. Transplanted SSCs demonstrated the earliest transference and colonization in recipient testes 7 days after transplantation. The newly generated germ cell layers appeared to be intact during spermatogenesis 90-days post-transplantation. This manual injection technique under microscope provides an alternative method to deliver the SSCs into the recipient seminiferous tubules.

In search of an efficient injection technique for future clinical application of spermatogonial stem cell transplantation: infusion of contrast dyes in isolated cadaveric human testes

To develop an efficient infusion technique for human spermatogonial stem cell transplantation. A mixture with ultrasonic contrast, computerized tomography (CT) contrast, and Chinese ink was injected into isolated human testes through different sites (the rete testis, the head of the epididymis, the deferent duct, and blind testicular infusion). Ultrasound transducer was used to visualize the injection site and to observe the flow of the mixture injected in the testes. Then, micro-CT scan was used to construct three-dimensional images, allowing the calculation of the testicular volume filled by the mixture. Finally the efficiency of the infusion was evaluated on histologic sections. Research laboratory. Cadaver testes obtained from autopsied bodies at the department of pathology. Ultrasound-guided infusion of contrast liquid. Contrast liquid-filled testis volume and presence of ink in seminiferous tubules. Ultrasonography clearly visualized the flow when seminiferous tubules were injected from the rete testis. No flow was observed when infusions were made either blindly, into the deferent duct, or into the head of the epididymis. On micro-CT no significant differences were observed between the different volumes. After rete testis infusion, ink particles were found in the lumen of the rete testis and in tubules, close and distant from the rete testis. A single ultrasound-guided injection of 800 μL in the rete testis may provide a promising method to transplant human spermatogonial stem cells in a clinical setting.

Stem cell transplantation for treating Duchenne muscular dystrophy: A Web of Science-based literature analysis.

OBJECTIVE:To identify global research trends in stem cell transplantation for treating Duchenne muscular dystrophy using a bibliometric analysis of Web of Science.DATA RETRIEVAL:We performed a bibliometric analysis of studies on stem cell transplantation for treating Duchenne muscular dystrophy from 2002 to 2011 retrieved from Web of Science.SELECTION CRITERIA:Inclusion criteria: (a) peer-reviewed published articles on stem cell transplantation for treating Duchenne muscular dystrophy indexed in Web of Science; (b) original research articles, reviews, meeting abstracts, proceedings papers, book chapters, editorial material, and news items; and (c) publication between 2002 and 2011. Exclusion criteria: (a) articles that required manual searching or telephone access; (b) documents that were not published in the public domain; and (c) corrected papers.MAIN OUTCOME MEASURES:(1) Annual publication output; (2) distribution according to subject areas; (3) distribution according to journals; (4) distribution according to country; (5) distribution according to institution; (6) distribution according to institution in China; (7) distribution according to institution that cooperated with Chinese institutions; (8) top-cited articles from 2002 to 2006; (9) top-cited articles from 2007 to 2011.RESULTS:A total of 318 publications on stem cell transplantation for treating Duchenne muscular dystrophy were retrieved from Web of Science from 2002 to 2011, of which almost half derived from American authors and institutes. The number of publications has gradually increased over the past 10 years. Most papers appeared in journals with a focus on gene and molecular research, such as Molecular Therapy, Neuromuscular Disorders, and PLoS One. The 10 most-cited papers from 2002 to 2006 were mostly about different kinds of stem cell transplantation for muscle regeneration, while the 10 most-cited papers from 2007 to 2011 were mostly about new techniques of stem cell transplantation for treating Duchenne muscular dystrophy.CONCLUSION:The publications on stem cell transplantation for treating Duchenne muscular dystrophy were relatively few. It also needs more research to confirm that stem cell therapy is a reliable treatment for Duchenne muscular dystrophy.

Biological Characteristics of Fish Germ Cells and their Application to Developmental Biotechnology

We have revealed several unique characteristics of germ cell development using rainbow trout, including the fact that spermatogonia transplanted into the peritoneal cavity of newly hatched embryos migrate toward recipient gonads, that spermatogonia transplanted into female recipients start oogenesis and produce functional eggs and that diploid germ cells transplanted into triploid trout can complete gametogenesis. By combining these unique features of fish germ cells, we established allogeneic and xenogeneic transplantation systems for spermatogonia in several fish species. Spermatogonia isolated from the mature testes of vasa-green fluorescent protein (Gfp) transgenic rainbow trout were transplanted into the peritoneal cavity of triploid masu salmon newly hatched embryos. These spermatogonia migrated toward recipient salmon genital ridges with extending pseudopodia and were subsequently incorporated into them. We further confirmed that the donor-derived spermatogonia resumed gametogenesis and produced sperm and eggs in male and female salmon recipients, respectively. By inseminating the resulting eggs and sperm, we obtained only rainbow trout offspring in the F1 generation, suggesting that the triploid salmon recipients produced functional gametes derived only from donor trout. We further confirmed that this intra-peritoneal transplantation of germ cells is applicable to several marine fishes, which could be of benefit in the production of bluefin tuna that has a large broodstock (>100 kg) and is difficult to maintain in captivity. Gamete production of bluefin tuna could be more easily achieved by generating a surrogate species, such as mackerel, that can produce tuna gametes.

Wnt/β-catenin signaling cell-autonomously converts non-hepatic endodermal cells to a liver fate

SummaryWnt/β-catenin signaling plays multiple roles in liver development including hepatoblast proliferation and differentiation, hepatocyte differentiation, and liver zonation. A positive role for Wnt/β-catenin signaling in liver specification was recently identified in zebrafish; however, its underlying cellular mechanisms are unknown. Here, we present two cellular mechanisms by which Wnt/β-catenin signaling regulates liver specification. First, using lineage tracing we show that ectopic hepatoblasts, which form in the endoderm posterior to the liver upon activation of Wnt/β-catenin signaling, are derived from the direct conversion of non-hepatic endodermal cells, but not from the posterior migration of hepatoblasts. We found that endodermal cells at the 4–6th somite levels, which normally give rise to the intestinal bulb or intestine, gave rise to hepatoblasts in Wnt8a-overexpressing embryos, and that the distribution of traced endodermal cells in Wnt8a-overexpressing embryos was similar to that in controls. Second, by using an endoderm-restricted cell-transplantation technique and mosaic analysis with transgenic lines that cell-autonomously suppress or activate Wnt/β-catenin signaling upon heat-shock, we show that Wnt/β-catenin signaling acts cell-autonomously in endodermal cells to induce hepatic conversion. Altogether, these data demonstrate that Wnt/β-catenin signaling can induce the fate-change of non-hepatic endodermal cells into a liver fate in a cell-autonomous manner. These findings have potential application to hepatocyte differentiation protocols for the generation of mature hepatocytes from induced pluripotent stem cells, supplying a sufficient amount of hepatocytes for cell-based therapies to treat patients with severe liver diseases.

Are dental X‐rays causing Alzheimer's? Ten reasons to take a closer look

The speed with which Alzheimer's disease has become a killer worldwide argues against a complex etiology because the more factors required for an outcome, the more difficult it is to meet all of the conditions necessary to cause it. In the United States, Alzheimer's is the sixth leading cause of death for people of any age, with fatalities increasing 66% over the same period of time that deaths due to other leading causes, such as stroke, heart disease and diabetes, decreased. While important genetic causes and associations to Alzheimer's have been made, the fact that these genetic factors do not always cause Alzheimer's and do not explain the majority of cases suggest that something else is directly responsible for Alzheimer's. Likewise, significant discoveries regarding Abeta plaques have indicated that plaques removal is not the key to slowing or stopping Alzheimer's. This presentation builds on ideas set forth in “Dental X-ray exposure and Alzheimer's disease: A hypothetical etiological association,” published online in March of 2011 in the Journal of Medical Hypotheses, which demonstrated that people in countries with the lowest Alzheimer's prevalence also had the least possible exposure to dental X-rays. The article also proposed a biological pathway by which ionizing radiation damaged microglia telomeres, causing microglia to prematurely lose their neuroprotective properties, leading to neuronal death. A unifying explanation will be presented for 10 distinctive Alzheimer's facts that are either not currently understood or are explained by separate means. If Alzheimer's dementia is caused by the collapse of the microglia population that supports neurons, urgent attention should be given to continuing work to perfect self-donated bone marrow or dental pulp stem cell transplant techniques to replenish microglia populations. This intervention has been suggested to be efficacious because it helps clear plaques, yet may be more important as a means of supporting and maintaining neurons while the original microglia population prematurely ages and dies.

How to Treat Osteochondritis Dissecans of the Knee: Surgical Techniques and New Trends

Osteochondritis dissecans is a relatively common cause of knee pain. The aim of this study was to describe the outcomes of five different surgical techniques in a series of sixty patients with osteochondritis dissecans. Sixty patients (age 22.4 ± 7.4 years, sixty-two knees) with osteochondritis dissecans of a femoral condyle (forty-five medial and seventeen lateral) were treated with osteochondral autologous transplantation, autologous chondrocyte implantation with bone graft, biomimetic nanostructured osteochondral scaffold (MaioRegen) implantation, bone-cartilage paste graft, or a "one-step" bone-marrow-derived cell transplantation technique. Preoperative and follow-up evaluation included the International Knee Documentation Committee (IKDC) score, the EuroQol visual analog scale (EQ-VAS) score, radiographs, and magnetic resonance imaging. The global mean IKDC score improved from 40.1 ± 14.3 preoperatively to 77.2 ± 21.3 (p < 0.0005) at 5.3 ± 4.7 years of follow-up, and the EQ-VAS improved from 51.7 ± 17.0 to 83.5 ± 18.3 (p < 0.0005). No influence of age, lesion size, duration of follow-up, or previous surgical procedures on the result was found. The only difference among the results of the surgical procedures was a trend toward better results following autologous chondrocyte implantation (p = 0.06). All of the techniques were effective in achieving good clinical and radiographic results in patients with osteochondritis dissecans, and the effectiveness of autologous chondrocyte implantation was confirmed at a mean follow-up of five years. Newer techniques such as MaioRegen implantation and the "one-step" transplantation technique are based on different rationales; the first relies on the characteristics of the scaffold and the second on the regenerative potential of mesenchymal cells. Both of these newer procedures have the advantage of being minimally invasive and requiring a single operation.

0711 細胞移植用肝実質細胞が受けるせん断流による損傷(OS7 生物・生体に閧する流れ,オーガナイズドセッション)

Hepatocyte transplantation is one of the cell transplantation techniques for the certain diseases of the liver.It has considerable potential as an alternative treatment of solid organ transplantation.In particular,it is indispensable treatment to control ammonia metabolism for a newborn baby who has liver dysfunctions.It is important to investigate the mechanical property of the hepatocyte while medical transactions.In this study,collagenase perfusion method was used for cell separation.Damage of the hepatocyte in shear flow were measured using trypan blue solution with a micro channel.As a result,hepatocyte injury situation was observed with optical microscope.The destruction conditions of the hepatocyte were clarified and the optimal infusion condition of the hepatocyte transplantation was estimated.

精原細胞の異種間移植法を用いた水産有用海産魚類における代理親魚技術の確立

精原細胞の異種間移植法を用いた水産有用海産魚類における代理親魚技術の確立

Prevention of Idiopathic Pneumonia Syndrome by Intra-Bone Marrow Injection of Donor Cells

Abstract 1902Several clinical and experimental studies have reported that a recently developed intra-bone marrow (IBM) stem cell transplantation (SCT) technique gives high rates of engraftment and is associated with a low incidence of acute graft-versus-host disease (GVHD). Idiopathic pneumonia syndrome (IPS) is a significant cause of mortality and remains a major obstacle after allogeneic SCT. In the present study, the extent of IPS after IBM-SCT was compared with that after conventional intravenous SCT (IV-SCT) using a lethally irradiated B6(H-2b) into F1 (H-2b/d) mouse IPS model.Compared with IV-SCT, IBM-SCT significantly improved the clinical GVHD score and reduced total and CD3+ T cell numbers in bronchoalveolar lavage fluid (9.6± 3.5 vs. 21.3 ± 0.5 x104/ml; p < 0.05). Histopathological examination of lung tissue at 6 weeks post-SCT showed significantly reduced IPS pathology in mice that underwent IBM-SCT. To explore the mechanisms of the reduction in IPS pathology in mice that underwent IBM-SCT, we monitored the in vivo distributions of infused donor cells and compared them between mice that underwent IBM-SCT versus IV-SCT. Recipient mice were imaged at different time points (1, 2, 3, and 6 h, and 1, 2, 3, and 5 days), using a lethally irradiated luciferase-expressing transgenic FVB/N (FVB/N luc+)(H-2q) into BALB/c (H-2d) mouse model. In vivo bioluminescence imaging (BLI) analysis revealed that the majority of injected donor cells were trapped in the lung 1 h after IV-SCT. In contrast, almost all donor cells were localized in the injected limbs 1 h after IBM-SCT, and significantly fewer cells had reached the lung (3.1± 0.7 vs. 16.7± 1.1 x105 photons/sec/animal, IBM-SCT vs. IV-SCT, p < 0.01; Figure). After syngeneic (FVB/N luc+ into FVB/N) SCT, the majority of the injected cells were also trapped in the lung 1 h after IV-SCT, and a similar difference was observed in donor cell distribution in the lung after IV-SCT versus IBM-SCT (2.4± 0.6 vs. 11.6± 1.3 x105 photons/sec/animal; p < 0.01). These results suggest that initial cell localization to the lung is dependent on the SCT method. At 2 days post-SCT, we examined the profiles of chemokines produced locally in the lung (CCL2, CCL3, CXCL1, CCL5, and CCL8). The mRNA expression of CC chemokines, especially CCL2, was more strongly induced in the lung after allogeneic IV-SCT than after allogeneic IBM-SCT (0.098 ± 0.020 vs. 0.020 ± 0.003 units/GAPDH mRNA; p < 0.05). A similar difference was observed between mice that underwent syngeneic IV-SCT and syngeneic IBM-SCT, suggesting that increases in chemokine levels in the lung early post-SCT are also dependent on the SCT method. At 5 days post-syngeneic SCT, BLI analysis revealed that no difference was observed in donor cell distribution in the lung after IV-SCT versus IBM-SCT (4.8± 1.1 vs. 4.6± 2.5 x107 photons/sec/animal; p =0.94). On the other hand, the BLI signals dramatically increased in the lungs of mice that had undergone allogeneic IV-SCT after day 2 post-SCT and there was a significant difference in the BLI signals between IV-SCT and IBM-SCT mice at 5 days post-SCT (50.9± 6.6 vs. 16.0± 6.2×107 photons/sec/animal; p < 0.05). These results suggest that increases in chemokine levels in the lung at day 2 post-SCT lead to increases in the allogeneic response in the lung.In summary, we have shown that the initial localization of donor cells to the lung and increases in lung chemokine levels are dependent on the SCT method. The targeting of donor cell trafficking to the lung may be a promising strategy for preventing IPS, and IBM-SCT may reduce IPS after allogeneic SCT. [Display omitted] Disclosures:No relevant conflicts of interest to declare.

Association Between Genetic Variants in Immune Response Genes and Outcomes After Hematopoietic Cell Transplantation

Abstract 3049 Background:Despite rapid advances in allogeneic hematopoietic cell transplant (HCT) techniques, the procedure continues to be associated with a high morbidity and mortality with transplant related mortality (TRM) ranging from 15% to as high as 50%. Acute graft versus host disease (GvHD) remains a major cause of morbidity and an important cause of early TRM in patients undergoing allogeneic HCT. Though variations in major histocompatibiliy (MHC) genes are well known determinants of acute GvHD, there is increasing evidence that genetic variation in immune/cytokine response pathways also contributes significantly to the pathogenesis of acute GvHD. Methods:We evaluated the association of single nucleotide polymorphisms (SNPs) in non-MHC dependent immune/cytokine response pathways (n= 77 SNPs in recipients and donors) with acute GvHD and TRM at one year in a cohort of 425 recipient-donor pairs who underwent allogeneic HCT for treatment of hematologic malignancies at the University of Minnesota between 1998 and 2007. We used a Fine and Gray proportional hazards model adjusted for competing risk to evaluate the association between the genetic variants and time to acute GvHD and time to TRM. Results:After adjustment for recipient age at transplant, race, diagnosis, disease status at transplant, gender mismatch, CMV serostatus of recipient and donor, recipient and donor sex, donor type, conditioning regimen (myeloablative or reduced intensity) and year of transplant, one recipient SNP, rs646005 in the TIRAP gene and one donor SNP, rs2232596 in the LBP gene were marginally associated with increased risk for grade II-IV acute GvHD (Hazard ratio (HR) = 1.33 (95% Confidence interval (CI): 1.07 – 1.62) and 1.32 (1.07 – 1.66) respectively; p =0.01). Similar analyses for TRM showed recipient SNP rs5498 in the ICAM1 gene was associated with a decreased risk of TRM (HR: 0.67(95% CI: 0.50 – 0.89); p=0.0006) while recipient SNPs rs1739654 in the LBP gene and rs3804100 in the TLR2 gene were associated with an increased TRM risk (HR: 1.86 (95% CI: 1.16 – 2.97) and 1.66 (95% CI: 1.11 – 2.48) respectively; p =0.01). Conclusions:These findings are consistent with previous reports that support a role for genetic variants in the innate immune response pathways among both recipients and donors can influence the risk of acute GvHD and TRM among patients undergoing allogeneic HCT. These findings indicate that a systematic search for genetic variants in the innate immune response pathways may identify novel biomarkers that can be used to identify patients at high risk for acute GvHD and TRM. Disclosures:Weisdorf:Genzyme: Consultancy, Research Funding.

Selective retrograde coronary venous perfusion with over-the-wire balloon: a new technique for cell transplantation

ObjectiveEffective cell homing to ischemic myocardium remains a practical challenge. This study evaluated the efficiency of cell delivery via coronary venous system using over-the-wire balloon in a canine model of...

Molecular pathology of tumor‐initiating cells: Lessons from Philadelphia chromosome‐positive leukemia

Recent improvements in cell purification and transplantation techniques have contributed to the identification of cell populations known as tumor-initiating cells (TIC). This discovery has led to the 'cancer stem cell hierarchy' concept, which holds that tumors are organized as a hierarchy of malignant tissues sustained by such TIC. However, this concept remains controversial. In this review, we examine recent advances in cancer stem cell research that have been generated from studies of Philadelphia (Ph) chromosome-positive leukemia. The abnormal Ph chromosome, which arises from a translocation creating the BCR-ABL1 fusion gene, is most commonly associated with chronic myelogenous leukemia (CML) and precursor B cell acute lymphoblastic leukemia (B-ALL). Examination of the pathophysiology of these diseases has provided interesting insights into not only the hierarchy of leukemia stem cells but also their clonal evolution. Both shared and unique regulatory mechanisms affecting normal and CML stem cell behavior have been identified. On the other hand, genetic diversity in specific clones of Ph(+) B-ALL that drive clonal evolution has recently come to light. Our expanding knowledge of the biology of TIC contributes to the progress of cancer research, and may open the door to new concepts in cancer therapy.

Incidence, risk factors, and clinical outcomes of cataracts following hematopoietic stem cell transplantation

To evaluate the characteristics of cataracts following hematopoietic stem cell transplantation (HSCT), 561 patients without any ocular complications before HSCT were reviewed. Ocular complications occurred in 196 patients (34.9%). Cataract was diagnosed in 45 of 561 patients. The 10-year estimated cataract incidence was 14%. The median onset time was 498 days after transplant (range, 38-4,535 days). The onset of cataracts was later than that of other ocular complications. Phacoemulsification was performed in 13 patients (29%) at a median of 1,028 days after HSCT (range, 375-4,549 days). In 6 of 13 patients (46%) who underwent cataract surgery, visual acuities gradually deteriorated because of the development of posterior capsule opacification. YAG (yttrium-aluminum-garnet) laser posterior capsulotomy successfully recovered their sight without any sequelae. A multivariate analysis revealed that age, acute and chronic graft versus host disease (GVHD), and systemic infection were significant risk factors for cataract, whereas neither steroid nor total body irradiation (TBI)-containing regimens affected the development of cataract. Our results showed that the clinical outcomes of cataract after HSCT are favorable and comparable to those in the general population. Hyperfractionated TBI with eye shielding may be effective for the prevention of cataract.

Management of Mustard Gas-Induced Limbal Stem Cell Deficiency and Keratitis

To report the clinical findings and compare outcomes of different surgical techniques evolved over time in a large series of patients with delayed-onset mustard gas keratitis (MGK). Retrospective, comparative, interventional case series. Ninety Iranian male survivors (175 eyes) of Iraqi chemical warfare with chronic or delayed-onset MGK. The symptoms and clinical findings of patients are presented, and medical and surgical interventions to address dry eye, limbal ischemia and limbal stem cell deficiency (LSCD), and corneal involvements are explained. The results of limbal stem cell transplantation techniques (living-related conjunctival-limbal allograft [lrCLAL] versus keratolimbal allograft [KLAL]) as well as corneal transplantation techniques (penetrating keratoplasty [PK] versus lamellar keratoplasty [LK]) are compared in terms of clinical outcomes and graft survival rates. Ocular findings and appropriate surgical approach for LSCD and corneal involvements. A total of 175 eyes of 90 cases (all male) between 34 and 68 years of age were followed up for 101±30.3 months (range, 36-198 months). The most common ocular involvements were chronic blepharitis and dry eye. Conjunctival vascular abnormalities and limbal ischemia were observed in 27.4% and 29.7% of eyes, respectively. Limbal stem cell deficiency necessitating stem cell transplantation developed in 41.1% of eyes. The most common corneal sign was central and peripheral anterior stromal opacity (58.9%), followed by corneal stromal thinning (36.0%) and neovascularization (27.4%). Living-related conjunctival-limbal allograft was performed in 32 eyes, and KLAL was performed in 40 eyes. The rejection-free graft survival rate was 39.1% in the lrCLAL group and 80.7% in the KLAL group at month 40, with a mean length of 24.9 and 68.8 months, respectively (P = 0.02). Thirty eyes underwent PK and 51 underwent LK. Corneal graft failure was observed in 9 PK eyes and in 6 LK eyes. The rejection-free graft survival rate was 39.0% in the PK group and 90.3% in the LK group at month 28, with a mean length of 29.6 and 85.0 months, respectively (P<0.001). Chemical warfare victims who initially have mild symptoms ultimately may experience significant ocular involvements requiring surgical intervention. Limbal and corneal abnormalities can be managed best by KLAL and LK, respectively.

Graft-versus-Host-Disease (GvHD) – ein Update

GvHD remains associated with significant morbidity and mortality despite new techniques for allogeneic stem cell transplantation (SCT), such as optimized conditioning regimens. Within the past ten years, the incidence of acute GvHD has remained unchanged and the incidence of chronic GvHD has even increased. The traditional classification of GvHD according to the time of clinical manifestation is now out-dated. Acute GvHD symptoms may even occur after 100 days; vice versa, primary chronic GvHD may already be observed one month after stem cell transplantation. The current classification introduced by the National Institutes of Health includes classic acute GvHD (up to 100 days), late-onset acute GvHD (after 100 days), as well as an overlap syndrome showing features of acute and chronic GvHD and classic chronic GvHD without any time limit. Diagnosis of GvHD of the skin remains difficult because of histological similarities to drug eruptions and viral exanthems. In this first part of the article the pathophysiology, classification, skin manifestations of acute and chronic GvHD and the histopathology will be presented. In a second part the prognosis, prophylaxis and therapy of GvHD will be discussed.

Everything is on the Head

Everything is on the Head

Osteoanagenesis after transplantation of bone marrow-derived mesenchymal stem cells using polyvinylidene chloride film as a scaffold

The aim of this study was to develop a new cell transplantation technique for osteoanagenesis at bone defect sites. Polyvinylidene chloride (PVDC) film was evaluated because of its good biocompatibility and flexibility. We used this film as both a cell scaffold and a barrier membrane. Initially, the cell compatibility of the PVDC film for fibroblast-like cells and osteoblast-like cells was confirmed. Subsequently, bone marrow cells were obtained from rats and cultured on PVDC films in two kinds of medium. The PVDC films with bone marrow-derived mesenchymal stem cells (MSCs) were then applied to critical-sized bone defects in the calvarial bone of rats. After the transplantation, the surgical sites were dissected out and evaluated by soft X-ray radiography, micro-CT analysis and histological examinations. The bone marrow-derived MSC-transplanted rats showed greater bone regeneration than the control rats. Therefore, PVDC film is considered to be useful as a scaffold for bone regeneration.

Restoration of spermatogenesis after transplantation of c-Kit positive testicular cells in the fowl

Transplantation of male germ line cells into sterilized recipients has been used in mammals for conventional breeding as well as for transgenesis. We have previously adapted this approach for the domestic chicken and we present now an improvement of the germ cell transplantation technique by using an enriched subpopulation of c-Kit-positive spermatogonia as donor cells. Dispersed c-Kit positive testicular cells from 16 to 17 week-old pubertal donors were transplanted by injection directly into the testes of recipient males sterilized by repeated gamma irradiation. We describe the repopulation of the recipient's testes with c-Kit positive donor testicular cells, which resulted in the production of functional heterologous spermatozoa. Using manual semen collection, the first sperm production in the recipient males was observed about nine weeks after the transplantation. The full reproduction cycle was accomplished by artificial insemination of hens and hatching of chickens.