Heterotopic Autotransplantation Research Articles

Abdominal peritoneal splenosis (clinical case)

Background. Splenosis, or heterotopic autotransplantation of spleen tissue, is a benign disease, occurring in patient after splenectomy. This formation demands correct differential diagnosis because it is often diagnosed as a tumor. Main methods of splenosis diagnosis are ultrasound inspection, CT, MRI, but detailed anamnesis is also important to set the correct diagnosis. The clinical case in this report is aimed to show importance of timely and correct diagnosis which will allow to avoid unnecessary invasive diagnostic procedures, late diagnosis and decreasing of medical assistance quality. Clinical case. patient 61 y. o. is suspected on a tumor in left iliac region. He noted the trauma after falling on a back. The inspection was performed: CT of iliac region showed formation in the small intestines. Splenectomy was carried out 20 years ago. The patient is set to the clinical medical center “Medicinsky gorod” to a surgeon-oncologist. physical examination and laboratory analysis showed no pathological changes. CT of abdominal organs showed formation with regular edges. MRI of pelvic organs with contrast agent showed formation accumulating contrast irregularly. Control CT of abdominal organs showed no increasing of formation size. Conclusion. Abdominal splenosis is a benign disease, occurring after spleen tissue implantation in the abdominal cavity after spleen trauma or splenectomy. Splenosis course is usually asymptomatic and it is often diagnoses as a peritoneal canceromatosis mistakenly. Splenosis has to be included in the differential diagnosis in patients with trauma or splenectomy in anamnesis.

Intrahepatic splenosis as a differential diagnosis for hepatic neoplasia in a dog

AbstractIntrahepatic splenosis is the heterotopic autotransplantation of splenic tissue secondary to splenic trauma or splenectomy. A 12‐year‐old Cairn terrier was referred due to lethargy, collapse and abdominal discomfort. Abdominal ultrasound and a computed tomography scan (CT) revealed an intrahepatic mass in the left medial liver lobe. The dog underwent surgery for liver lobectomy, and subsequent histopathology revealed intrahepatic splenosis. The dog had undergone splenectomy 3 years previously due to splenic rupture following an abdominal trauma.

Radiological study of splenosis: A great mimicker of tumor a literature review with case reports

Splenosis is a harmless condition after a splenic injury or a spleen removal. It is heterotopic autotransplantation of splenic tissue seen inside the abdominal and pelvic holes. On imaging, various conditions can be differential diagnosis with splenosis, including metastatic infection, peritoneal carcinoma, peritoneal mesothelioma, stomach lymphoma, renal disease, hepatic adenomas, or endometriosis, dependent on its transmission. This review aims to increase public knowledge of this condition to avoid needless biopsies, surgeries, or treatments. Methods: We searched the PubMed and Scopus databases for relevant literature from 2002 through 2022 using the Medical Subject Headings Splenosis, Radiological, and Radionuclide diagnostic tools. In total, Thirty-five cases of splenosis were identified and reviewed. Conclusion: Splenosis should always be considered a differential diagnosis for soft tissue nodules in the abdomen and pelvis in patients with splenic surgery or trauma, especially without systemic symptoms. The literature documented only a few examples of splenosis, along with the radiologic features of these lesions. We review instances of splenosis identified by Scintigraphy, magnetic resonance imaging, computed tomography, and ultrasound. Our research expects this review to be considered a unique to compare the diagnosis made by radiologic and Radionuclide testing without any other form of treatment.

Effects of different subcutaneous sites on heterotopic autotransplantation of canine ovarian tissue.

Ovarian tissue transplantation makes it possible to restore fertility; however, the success of this technique depends on the transplant region used. Therefore, this study aimed to evaluate the effect of two subcutaneous regions on canine ovarian transplantation, pinna (Pi) and neck (Ne), for 7 and 15 days. Ovaries collected by ovariosalpingohysterectomy were fragmented using a punch device. Fresh fragments were fixed, and the others were immediately grafted onto the animal itself in the Pi and Ne regions for 7 and 15 days. Recovered fragments were evaluated for histology (morphology, development and stromal density), picrosirius (collagen fibers), and immunohistochemistry (fibrosis and cell proliferation). The results showed that follicular normality rates were lower in Pi-7 (78%) vs. control (90%) and Pi-15 (86%), similar in Ne-7 (92%) and superior in Ne-15 (97%) compared to the control, with the effect of the region Ne (94%) superior (P < 0.05) to Pi (82%). Stromal density reduced in both regions vs. control but was similar within 15 days. Fragments from both regions showed higher fibronectin labeling and deposition of type I and lower type III collagen fibers (P < 0.05) vs. control. Proliferation rates in Ne-7 were higher (P < 0.05) than in control, and Pi-15 was higher (P < 0.05) than Ne-15. In conclusion, the pinna may be a region with greater potential than the neck after a 15-day autotransplantation of canine ovarian tissue.

Spleen regeneration after subcutaneous heterotopic autotransplantation in a mouse model

BackgroundSplenectomy may lead to severe postoperative complications, including sepsis and cancers. A possible solution to this problem is heterotopic autotransplantation of the spleen. Splenic autografts rapidly restore the regular splenic microanatomy in model animals. However, the functional competence of such regenerated autografts in terms of lympho- and hematopoietic capacity remains uncertain. Therefore, this study aimed to monitor the dynamics of B and T lymphocyte populations, the monocyte-macrophage system, and megakaryocytopoiesis in murine splenic autografts.MethodsThe model of subcutaneous splenic engraftment was implemented in C57Bl male mice. Cell sources of functional recovery were studied using heterotopic transplantations from B10-GFP donors to C57Bl recipients. The cellular composition dynamics were studied by immunohistochemistry and flow cytometry. Expression of regulatory genes at mRNA and protein levels was assessed by real-time PCR and Western blot, respectively.ResultsCharacteristic splenic architecture is restored within 30 days post-transplantation, consistent with other studies. The monocyte-macrophage system, megakaryocytes, and B lymphocytes show the highest rates, whereas the functional recovery of T cells takes longer. Cross-strain splenic engraftments using B10-GFP donors indicate the recipient-derived cell sources of the recovery. Transplantations of scaffolds populated with splenic stromal cells or without them afforded no restoration of the characteristic splenic architecture.ConclusionsAllogeneic subcutaneous transplantation of splenic fragments in a mouse model leads to their structural recovery within 30 days, with full reconstitution of the monocyte-macrophage, megakaryocyte and B lymphocyte populations. The circulating hematopoietic cells provide the likely source for the cell composition recovery.

Xenograft model of heterotopic transplantation of human ovarian cortical tissue and its clinical relevance.

Xenografts of human ovarian cortical tissue provide a tractable model of heterotopic autotransplantation that is used for fertility preservation in patients undergoing ablative chemo/radiotherapy. This study describes the behavior of hundreds of xenografts to establish a framework for the clinical function of ovarian cortex following autotransplantation over short- and long-term intervals. More than 200 live births have been achieved using autotransplantation of cryopreserved ovarian cortical fragments, yet challenges remain to be addressed. Ischemia of grafted tissue undermines viability and longevity, typically requiring transplantation of multiple cortical pieces; and the dynamics of recruitment within a graft and the influence of parameters like size and patient age at the time of cryopreservation are not well-defined. Here, we describe results from a series of experiments in which we xenografted frozen/thawed human ovarian tissue (n = 440) from 28 girls and women (age range 32 weeks gestational age to 46 years, median 24.3 ± 4.6). Xenografts were recovered across a broad range of intervals (1-52 weeks post-transplantation) and examined histologically to quantify follicle density and distribution. The number of antral follicles in xenografted cortical fragments correlated positively with the total follicle number and was significantly reduced with increased patient age. Within xenografts, follicles were distributed in focal clusters, similar to the native ovary, but the presence of a leading antral follicle coincided with increased proliferation of surrounding follicles. These results underscore the importance of transplanting ovarian tissue with a high density of follicles and elucidate a potential paracrine influence of leading antral follicles on neighboring follicles of earlier stages. This temporal framework for interpreting the kinetics of follicle growth/mobilization may be useful in setting expectations and guiding the parameters of clinical autotransplantation.

Extensive intrathoracic and intraperitoneal splenosis mimicking mesothelioma: a case report

BackgroundSplenosis is the heterotopic autotransplantation of splenic tissue after severe splenic trauma and/or splenectomy. The epidemiology is elusive, but splenosis is frequently misdiagnosed as malignant tumors of gastrointestinal, gynecological, or hematological origin before the correct diagnosis is ultimately found. We herein report a rare case of combined, extensive intraabdominal and intrathoracic splenosis initially presenting as pleural mesothelioma.Case presentationA 63-year-old Caucasian male presented with dyspnea and recurring thoracic pain. Initial X-ray and computed tomography scans showed disseminated intrathoracic and intraabdominal lesions. Consequently, thoracoabdominal mesothelioma or a polytopically metastasized cancer of unknown origin was suspected. A thorough examination of the patient’s medical history and contrast-enhanced ultrasound by a skilled examiner revealed the diagnosis of extensive abdominal and thoracic splenosis as a consequence of an abdominal gunshot wound with a ruptured diaphragm several decades earlier. Timely diagnosis by noninvasive measures prevented the patient from potential complications of harmful diagnostic procedures, including nuclear imaging and biopsies. The patient is currently treated for hepatitis C and chronic obstructive lung disease, whereas no specific treatment for splenosis is required.ConclusionsWe present a case of rare intrathoracic and intraperitoneal splenosis mimicking mesothelioma. Contrast-enhanced ultrasound and thorough patient history were used for diagnosis and prevented this patient from having to undergo potentially harmful diagnostics. Splenosis can occur after splenic trauma and, consequently, needs to be considered as a rare differential diagnosis to malignant tumors of various origins when a matching patient history is obtained.

L-Carnitine improves follicular survival and function in ovarian grafts in the mouse.

Ovarian tissue transplantation is performed to preserve fertility in patients undergoing chemotherapy and radiotherapy. However, the ischemia-reperfusion injury which occurs after the ovarian tissue transplantation causes follicular depletion and apoptosis. l -Carnitine has antioxidant and anti-inflammation properties. Therefore, we aimed to investigate the beneficial effect of l -carnitine on mouse ovaries following heterotopic autotransplantation. Mice were randomly divided into three groups (six mice per group): control, autografted and autografted+l -carnitine (200mg/kg daily intraperitoneal injections). Seven days after ovary autografting, the serum levels of malondialdehyde (MDA), total antioxidant capacity, tumor necrosis factor alpha (TNF-α), interleukin (IL)-6 and IL-10 were measured. Ovary histology, serum concentrations of progesterone and estradiol were also measured 28days after autotransplantation. Data were analysed using one-way analysis of variance (ANOVA) and Tukey test, and the means were considered significantly different at P Key results: In the autografted+l -carnitine group, the total volume of the ovary, the volume of the cortex, the number of follicles, the serum concentrations of IL-10, estradiol and progesterone significantly increased compared to the autografted group. In the autografted+l -carnitine group, serum concentrations of IL-6, TNF-α and MDA were significantly decreased compared to the autografted group. Our results indicated that l -carnitine can ameliorate the consequences of ischemia-reperfusion on the mice ovarian tissue following autotransplantation. l -carnitine improves the structure and function of transplanted ovaries.

New Experimental Model of Heterotopic Spleen Auto Transplantation

The aim of the study was to develop a new experimental model of heterotopic autotransplantation of the spleen by implanting splenic fragments into subcutaneous fat. Research Material: Experimental studies were conducted on 8 white mongrel rats weighing 300-400 grams at the age of 3 to 4 months. Research Results: Splenectomy is accompanied by significant violations of hemostasis and the immune system of the body, which justifies the feasibility of performing heterotopic autotransplantation of the spleen (GATS). In cases of infection or necrosis of the implant when performing traditional GATS methods, repeated surgical intervention under general anesthesia by laparotomy is required. The developed new method of GATS, which consists in implanting fragments of the spleen into subcutaneous adipose tissue, enveloped by an omentum "on the leg", in cases of necrosis or infection of the allograft, allows you to limit yourself to minimal surgical intervention under local anesthesia.

Heterotopic autotransplantation of equine ovarian tissue using intramuscular versus subvulvar grafting sites: Preliminary results

Ovarian tissue transplantation (OTT) is a technique well established and successfully applied in humans using mainly orthotopic or heterotopic transplantation sites. In livestock, OTT is still in its infancy and, therefore, different aspects of the technique, including the efficiency of different heterotopic OTT sites as well as the potential effect of age (i.e., young vs. old mares) in the ovarian graft quality, need to be investigated. The present study investigated the efficacy of the intramuscular (IM) or the novel subvulvar mucosa (SV) heterotopic autotransplantation sites to maintain the survivability of the grafts for 3 and 7 days post-OTT. Ovarian biopsy fragments were obtained in vivo and distributed to the following treatments: Fresh control group (ovarian fragments immediately fixed), SV-3, IM-3, SV-7, and IM-7. During and after graft harvesting, the macroscopic characteristics of the grafts (i.e., adherence, morphology, and bleeding) were scored, and the percentages of morphologically normal and developing preantral follicles as well as the follicular and stromal cell densities of the grafts were evaluated. The results were that similar (P > 0.05) macroscopic scores were observed between both transplantation sites 7 days post-OTT, with positive correlations (P < 0.01) found among adherence, morphology, and bleeding of the grafts. A lower (P < 0.05) percentage of morphologically normal follicles was found 7 days post-OTT in the SV site (82%) compared with the Fresh control group (99%) and IM site (95%); however, the percentages of developing follicles were similar (P > 0.05) between both transplantation sites 7 days post-OTT (30–43%). Although similar (P > 0.05) follicular densities were found in both transplantation sites in young and old mares at 3 and 7 days post-OTT, large individual variation in the follicular depletion rate was observed regardless of transplantation site. The Fresh control group and SV-7 treatments had higher (P < 0.05) stromal cell densities in young and old mares compared with both IM-7 treatments. When comparing transplant sites between young and old mares, the follicular density in old mares and the stromal cell density in young mares were greater (P < 0.05) in the SV than in the IM site. In conclusion, even though the transplantation sites differentially affected some end points, overall comparable findings of the OTT technique using both heterotopic autotransplantation sites (i.e., IM and SV) for equine ovarian tissue were observed.

Technique for Spleen Resection Using Radiofrequency Ablation

From modern positions, removal of the spleen is undesirable due to the risk of post-splenectomy syndrome development, where first of all, the clinical manifestations of the syndrome of suppressive post-splenectomy infections are taken into account, which justifies either performing organ-preserving surgical interventions on the spleen, or heterotopic autotransplantation of spleen tissue. Spleen resection is accompanied by significant blood loss, both during the intervention and in the postoperative period. The various methods of prevention of bleeding (clips, coagulation instruments, local hemostatics, etc.) are not always effective. Currently, there have been reports of the possibility of using radiofrequency ablation (RFA) in interventions on the spleen. The literature data and own 10 observations of spleen resection with the use of RFA in patients with focal organ lesions are presented. All described in literature technologies use the Rita® device for RF destruction and the corresponding attachment, which consists of 4 needle active electrodes. The spleen parenchyma is "burned" blindly, and the organ parenchyma is divided in the middle, between the treated injections. A new technique for performing spleen resection using radiofrequency ablation has been proposed at A.V. Vishnevsky National Medical Research Center of Surgery. The essence of our proposed method for minimizing blood loss during spleen resection is as follows: RFA-exposure is carried out along the zone of organ ischaemia under the control of ultrasound. For RFA, we used a Radionics Cool-Tip® Ablation System and a set of water-cooled electrodes (MEDTRONIC, USA). The features of the surgical intervention are noted. No bleeding was noted either intraoperatively or in the postoperative period in any case. A comparative analysis of the results of various methods of organ-saving surgical treatment is presented.

INTRAPERITONELA MULTINODULAR SPLENOSIS - AN INCIDENTAL EMERGENT LAPAROTOMY FINDING: A RARE CASE REPORT.

Splenosis is a rare benign condition that arises from the heterotopic autotransplantation of splenic tissue following iatrogenic or accidental splenic trauma. The aim of this publication is to present a rare case of multinodular intraperitoneal splenosis as an incidental finding at the time of emergent laparotomy, with a gap of more than 51 years after previous abdominal trauma.

Heterotopic autotransplantation of ovarian tissue in a large animal model: Effects of cooling and VEGF.

Heterotopic and orthotopic ovarian tissue autotransplantation techniques, currently used in humans, will become promising alternative methods for fertility preservation in domestic and wild animals. Thus, this study describes for the first time the efficiency of a heterotopic ovarian tissue autotransplantation technique in a large livestock species (i.e., horses) after ovarian fragments were exposed or not to a cooling process (4°C/24 h) and/or VEGF before grafting. Ovarian fragments were collected in vivo via an ultrasound-guided biopsy pick-up method and surgically autografted in a subcutaneous site in both sides of the neck in each mare. The blood flow perfusion at the transplantation site was monitored at days 2, 4, 6, and 7 post-grafting using color-Doppler ultrasonography. Ovarian grafts were recovered 7 days post-transplantation and subjected to histological analyses. The exposure of the ovarian fragments to VEGF before grafting was not beneficial to the quality of the tissue; however, the cooling process of the fragments reduced the acute hyperemia post-grafting. Cooled grafts compared with non-cooled grafts contained similar values for normal and developing preantral follicles, vessel density, and stromal cell apoptosis; lower collagen type III fibers and follicular density; and higher stromal cell density, AgNOR, and collagen type I fibers. In conclusion, VEGF exposure before autotransplantation did not improve the quality of grafted tissues. However, cooling ovarian tissue for at least 24 h before grafting can be beneficial because satisfactory rates of follicle survival and development, stromal cell survival and proliferation, as well as vessel density, were obtained.

S2932 Ectopic Spleen Tissue Mimicking Gastrointestinal Stromal Tumor, a Case Report of a Gastric Subepithelial Lesion

INTRODUCTION: Gastric subepithelial lesions (SEL) are found as often as 1 in 300 endoscopies. Indolent growing gastric splenosis mimicking a gastrointestinal stromal tumor (GIST) is rare. We present a case of a patient with a gastric SEL with growth concerning for a GIST that ultimately was diagnosed as ectopic splenic tissue (splenosis). This case highlights the importance of considering splenosis as a possible etiology for enlarging gastric SEL in patients with prior splenectomy. CASE DESCRIPTION/METHODS: A 52-year-old female with a remote history of solid pseudopapillary neoplasm (SPN) of the pancreas who received distal pancreatectomy and splenectomy 15 years ago presented with dyspepsia. 10 years ago, an esophagogastroduodenoscopy (EGD) revealed a 10-mm SEL in the gastric fundus. On endoscopic ultrasound (EUS) the lesion was described as a hypoechoic mass in the muscularis propria. Fine needle aspiration (FNA) was not performed given its challenging location. The patient opted for 4 years of annually computed tomography surveillance with stability of the lesion which was thought to be an accessory spleen. However, one year ago she presented to our clinic with dyspepsia. EUS revealed growth of the mass to 22.5 × 13.5 mm in the fourth layer with minimal vascularity suggestive of a GIST. Again, FNA was unsuccessful given its location in the greater curvature of the gastric fundus. Exploratory laparotomy was performed, the mass was excised, and pathology showed benign splenic tissue, consistent with splenosis. DISCUSSION: Splenosis and accessory spleen were considered initially as differential diagnoses in our patient given her splenectomy history. However, the indolent growth of the SEL and EUS appearance raised the concern of GIST. It is imperative to distinguish a GIST from other types of tumors to assess its malignant potential and treatment. Surgical excision was ultimately performed in our patient and the diagnosis of splenosis was made. Splenosis is a subtype of ectopic splenic tissue differentiated from accessory spleen, in that splenosis is acquired heterotopic autotransplantation and implantation of splenic tissue (usually after splenic trauma or splenectomy) while accessory spleen is a congenital anomaly with splenic arterial supply. Clinicians should be aware of the growth potential of splenosis as a mimicker of GISTs and other gastric SELs and consider ectopic splenic tissue a cause of SEL in patients without spleen.Figure 1.: EGD findings: Subepithelial nodule in the gastric fundus (year 0 vs year 9).Figure 2.: EUS findings (year 9): 22.5 × 13.5 mm well-defined hypoechoic and homogeneous mass arising from the fourth layer.Figure 3.: Histopathologic findings of subepithelial lesion: Benign splenic tissue, consistent with splenosis. (Hematoxylin-eosin x 100).

MITOCHONDRIAL METABOLISM IS PRESERVED FOLLOWING NORMOTHERMIC EX-VIVO KIDNEY PERFUSION OF GRAFTS PROCURED FOLLOWING CARDIAC DEATH

Background: Normothermic ex-vivo kidney perfusion (NEVKP) preservation has demonstrated superior graft outcomes for kidneys procured following donation-after-cardiac death (DCD) over static cold storage (SCS). To determine the mechanisms for this advantage, we compared the transcriptome from both groups through an unbiased genome-wide microarray analysis. Methods: Kidneys from 30kg Yorkshire pigs were subjected to 30min of warm ischemia and then 8hrs of pressure-controlled NEVKP or SCS prior to heterotopic autotransplantation. Renal biopsies were collected on POD3 and RNA transcript expression was determined utilizing the Affymetrix GeneChip® Porcine Gene 1.0 ST Array platform examining over 23,000 transcripts. Gene set enrichment analysis (GSEA) was completed using the Hallmark gene sets and Gene Ontology (GO) pathways. Results: NEVKP-stored grafts demonstrated parameters associated with improved post-transplant graft function including lactate clearance (0hr:10.29+/-0.48mmol/L vs 8hr:1.67+/-0.67mmol/L,n=5,p<0.01), decreasing intra-renal resistance (0hr:1.63+/-0.20mmHg/mL/min vs 8hr:0.41+/-0.13 mmHg/mL/min,n=5,p<0.01), and continuous urine production. Graft function was significantly improved with NEVKP compared to SCS following transplantation with lower peak serum creatinine (POD1:4.0+/-1.15mg/dL vs POD3:12.0+/-0.78mg/dL,n=5,p<0.01) and higher creatinine clearance on POD3 (39.6+/-11.8mL/min vs 2.6+/-0.9ml/min,n=5,p<0.01). GSEA demonstrated 11 Hallmark Gene Sets enriched in NEVKP compared to SCS including sets associated with fatty-acid metabolism and oxidative phosphorylation, while 7 Gene Sets were enriched in SCS compared to NEVKP (FDR-value<0.25,p<0.05). GO analysis demonstrated pathways associated with lipid oxidation/metabolism, the Krebs cycle, and pyruvate metabolism enriched in NEVKP vs SCS (FDR-value<0.05) Conclusions: NEVKP maintained or enriched transcripts of key mitochondrial metabolic pathways compared to SCS in grafts procured following DCD, likely accounting for the improved post-transplant graft function.

Significant Dysfunction of Kidney Grafts Exposed to Prolonged Warm Ischemia Is Minimized Through Normothermic Ex Vivo Kidney Perfusion.

Kidneys from 30-kg Yorkshire pigs were removed following 120 minutes of warm ischemia (WI). These grafts were then preserved in static cold storage (SCS, n = 6) or subjected to NEVKP (n = 5) for 8 hours before heterotopic autotransplantation. SRD was defined as postoperative day (POD) 4 oliguria <500 mL/24 h with serum K +> 6.0 mmol/L. All 4 surviving animals with 120 minutes WI grafts stored with SCS developed SRD, compared with 1/5 in the NEVKP group (P = 0.02). The NEVKP group, when compared with SCS, also demonstrated significantly decreased serum creatinine peak values (1118.51 ± 206.90 µmol/L versus 1675.56 ± 98.15 µmol/L; P = 0.002) and higher creatinine clearance (POD4: 9.05 ± 6.97 mL/min versus 0.89 ± 0.56 mL/min; P = 0.05). By POD7, serum creatinine was not significantly different than baseline in the NEVKP (431.49 ± 492.50 µmol/L versus 90.19 ± 14.15 µmol/L, respectively; P = 0.20) but remained elevated following SCS (1189.25 ± 309.47 µmol/L versus 97.26 ± 29.18 µmol/L, respectively; P < 0.01). Histology demonstrated significantly decreased tubular injury scores compared with SCS grafts (P = 0.03). Kidney grafts subjected to 120 minutes WI before retrieval showed significant improvement in function, prevention of SRD, and decreased injury following 8 hours of NEVKP.

Fibrin Scaffold Incorporating Platelet Lysate Enhance Follicle Survival and Angiogenesis in Cryopreserved Preantral Follicle Transplantation.

Background: Transplantation of cryopreserved follicles can be regarded as a promising strategy for preserving fertility in cancer patients under chemotherapy and radiotherapy by reducing the risk of cancer recurrence. The present study aimed to evaluate whether fibrin hydrogel supplemented with platelet lysate (PL) could be applied to enhance follicular survival, growth, and angiogenesis in cryopreserved preantral follicle grafts. Materials and Methods: Preantral follicles were extracted from 15 four-week-old NMRI mice, cryopreserved by cryotop method, and encapsulated in fibrin-platelet lysate for subsequent heterotopic (subcutaneous) auto-transplantation into the neck. Transplants were assessed in three groups including fresh follicles in fibrin-15%PL, cryopreserved follicles in fibrin-15%PL, and cryopreserved follicles in fibrin-0% PL. Two weeks after transplantation, histological, and immunohistochemistry (CD31) analysis were applied to evaluate follicle morphology, survival rate, and vascular formation, respectively. Results: Based on the results, fibrin-15% PL significantly increased neovascularization and survival rate (SR) both in cryopreserved (SR=66.96%) and fresh follicle (SR=90.8%) grafts, compared to PL-less fibrin cryopreserved transplants (SR=28.46%). The grafts supplemented with PL included a significantly higher percentage of preantral and antral follicles. Also, no significant difference was observed in the percentage of preantral follicles between cryopreserved and fresh grafts of fibrin-15% PL. However, a significantly lower (P=0.03) percentage of follicles (23.37%) increased to the antral stage in cryopreserved grafts of fibrin-15%PL, compared to fresh grafts (35.01%). Conclusion: The findings demonstrated that fibrin-PL matrix could be a promising strategy to improve cryopreserved follicle transplantation and preserve fertility in cancer patients at the risk of ovarian failure.

Fibrin Scaffold Incorporating Platelet Lysate Enhance Follicle Survival and Angiogenesis in Cryopreserved Preantral Follicle Transplantation

Background: Transplantation of cryopreserved follicles can be regarded as a promising strategy for preserving fertility in cancer patients under chemotherapy and radiotherapy by reducing the risk of cancer recurrence. The present study aimed to evaluate whether fibrin hydrogel supplemented with platelet lysate (PL) could be applied to enhance follicular survival, growth, and angiogenesis in cryopreserved preantral follicle grafts. Materials and Methods: Preantral follicles were extracted from 15 four-week-old NMRI mice, cryopreserved by cryotop method, and encapsulated in fibrin-platelet lysate for subsequent heterotopic (subcutaneous) auto-transplantation into the neck. Transplants were assessed in three groups including fresh follicles in fibrin-15%PL, cryopreserved follicles in fibrin-15%PL, and cryopreserved follicles in fibrin-0% PL. Two weeks after transplantation, histological, and immunohistochemistry (CD31) analysis were applied to evaluate follicle morphology, survival rate, and vascular formation, respectively. Results: Based on the results, fibrin-15% PL significantly increased neovascularization and survival rate (SR) both in cryopreserved (SR=66.96%) and fresh follicle (SR=90.8%) grafts, compared to PL-less fibrin cryopreserved transplants (SR=28.46%). The grafts supplemented with PL included a significantly higher percentage of preantral and antral follicles. Also, no significant difference was observed in the percentage of preantral follicles between cryopreserved and fresh grafts of fibrin-15% PL. However, a significantly lower (P=0.03) percentage of follicles (23.37%) increased to the antral stage in cryopreserved grafts of fibrin-15%PL, compared to fresh grafts (35.01%). Conclusion: The findings demonstrated that fibrin-PL matrix could be a promising strategy to improve cryopreserved follicle transplantation and preserve fertility in cancer patients at the risk of ovarian failure. [GMJ.2020;9:e1558]

Radionuclide Liver-Spleen Scan to Detect Splenosis

Splenosis is defined as heterotopic auto-transplantation of splenic tissue at ectopic location following splenic injury or surgery. It can be seen in any location in the abdomen and thorax as well as rare sites such as the brain and subcutaneous tissue and derives its blood supply from adjoining structures. Accessory spleen is a congenital condition and draws its blood supply from accessory splenic artery. It is located in gastrosplenic or lienorenal ligament. They are fewer in number and without preceding trauma. Colloid liver-spleen scan objectively localizes splenosis. Presented here is a case of hemolytic anemia who underwent splenectomy in 2007 for hereditary spherocytosis. Patient had vague abdominal pain 12 years post-surgery. Sonography and CT scan showed soft tissue lesions in abdomen. Colloid liver-spleen scan showed evidence of reticuloendothelial cells confirming the diagnosis of splenosis.

A Systematic Study on Reproductive Endocrine Function Recovery From Subcutaneous Ovarian Autotransplantation in Mice After 2 Weeks

BackgroundHeterotopic ovarian transplantation can not only restore the reproductive and endocrine function of animals but can also be studied with a specialized animal model. The aim of the study was to determine whether the reproductive endocrine function of the subcutaneously transplanted ovary was restored after 2 weeks. MethodsThe ovaries of 7-week-old mice were autologously transplanted into the back muscle. Fourteen days later, the ovarian structure was examined by hematoxylin and eosin staining. We continuously observed vaginal smears for changes in the estrous cycle. Estrogen and androgen concentrations were detected on the 14th day. The oocytes were collected and then used for in vitro maturation (IVM) and in vitro fertilization (IVF). ResultsThe cyclical estrous cycle changes were similar to those of the control group. There were no differences in the serum androgen and estrogen levels between the graft and control groups. The oocytes were able to develop into blastocysts after IVM and IVF. These results indicated that ovarian endocrine and reproductive function were restored within 2 weeks. ConclusionOur studies have shown that this ovarian heterotopic autotransplantation technique is able to restore steroidogenic and gametogenic functions at day 14 after transplantation. So far, the 14th day after transplantation is a landmark during the recovery from autologous heterotopic ovarian transplantation in the back of the mouse. This time point is the appropriate window to study heterotopic ovarian transplantation in mice.