OBJECTIVETo observe the homing and differentiation of marrow-derived mesenchymal stem cells (MSC) transplanted intravenously in smoke inhalation injured rabbits.

METHODSThirty-two New Zealand big ear rabbits were divided into normal control group (NC), inhalation injury group (II), normal control + MSC treatment group (NM), and MSC treatment group (MT) according to the random number table, with 8 rabbits in each group. Rabbits in NC group were injected with 10 mL phosphate buffered saline (PBS) via ear marginal vein. Rabbits in NM group were injected with 10 mL PBS containing the third generation MSC labeled by BrdU (1 × 10(7) per 10 mL PBS) via ear marginal vein. Severe smoke inhalation injury model was reproduced in the other two groups, among them rabbits in II group were treated as rabbits in NC group, rabbits in MT group treated as rabbits in NM group. On the 7th and 28th day post treatment (PTD), lung tissue and trachea tissue were harvested from four groups for observation on injury with HE staining. Homing of MSC in injured tissue was observed with immunohistochemistry staining. The differentiation of MSC into functional cells was observed with immunohistochemical double staining of combining nuclear marker BrdU with lung (trachea) membrane-specific marker aquaporin-5 (AQP-5), alkaline phosphatase (AKP), CD34, and cytokeratin respectively.

RESULTS(1) MSC homing in lung and trachea tissue was observed in MT group on PTD 7, which was not observed in NM group. (2) AQP-5, AKP, and CD34 positive MSC were observed in lung tissue in MT group on PTD 28, while cytokeratin positive MSC was not observed in trachea tissue. No positively marked MSC was observed in NM group. (3) Injury in lung and trachea was less severe in MT group than in II group; and the proliferation of fibroblasts was less in MT group.

CONCLUSIONSIntravenous injection of MSC to rabbits with smoke inhalation injury can migrate to lung and trachea tissue at obviously inflammatory site, and differentiate into alveolar epithelial cells typeI and II, and pulmonary vascular endothelial cells, which may participate in the process of tissue repair in smoke inhalation injury.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Endothelial Cells ; cytology ; Epithelial Cells ; cytology ; Lung ; cytology ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Pulmonary Alveoli ; cytology ; Rabbits ; Smoke Inhalation Injury ; pathology ; Trachea ; cytology

OBJECTIVETo follow the fate of murine epidermal stem cells (ESCs) seeded in a syngeneic dermal equivalent implanted in vivo.

METHODSEmbryonic stem (ES) cells were induced in vitro to differentiate into ESCs. After stained with a fluorescent dye Hoechst 33342, these ESCs were seeded into a fibroblast-collagen-gelatin sponge complex, functioning as a dermal equivalent model, and implanted subcutaneously into 129/J mice, which were syngeneic to these stem cells. The fate of these cells was observed with HE staining, immunocytochemical staining or Van Gieson's staining.

RESULTSThese ESCs were clearly visible in the implant by fluorescent microscopy 3 weeks or longer after implantation. These cells remained viable, differentiated into hair follicle-like structure, glandular structure, and gave rise to additional structures displaying features resembling native dermis. A number of markers were expressed in the differentiated structures, including CD29 (integrin beta1 subunit) and cytokeratin 18 (CK18). No apparent rejection or severe side effects were observed at least during the 10 weeks following implantation.

CONCLUSIONNow that ESCs could survive in vivo in this dermal equivalent model, and differentiate into hair follicle-like structures as well as glandular structures, it is feasible to use these cells as seed cells in the study to fabricate dermal equivalent having the potential to develop dermal appendages.

Animals ; Cell Culture Techniques ; Cells, Cultured ; Dermis ; cytology ; transplantation ; Embryonic Stem Cells ; cytology ; Epithelial Cells ; cytology ; Mice ; Mice, Inbred Strains ; Skin Transplantation ; Stem Cell Transplantation ; Stem Cells ; cytology ; Tissue Engineering

BACKGROUNDRe-epithelialization has remained a major obstacle in both tracheal and lung transplantations. This study examines the realization of re-epithelialization by epithelial inoculation in a rat heterotopic tracheal transplantation model.

METHODSThe original epithelia of tracheas from donor Wistar rats were removed and the tracheas were then inoculated with 10(6)/ml in vitro cultured epithelial cells of the Spraque-Dawley (SD) rat phenotype. These allo-tracheas were then heterotopically transplanted into SD rats. After 28 days, the allo-trachea tissues were recovered and assessed for epithelial morphology and cellular differentiation using immunohistochemical analysis. An additional experimental group was used to compare the outcomes of re-epithelialization in immunosuppressed animals.

RESULTSHistological examination showed that allografts with epithelial inoculation maintained patent tracheal lumens, which were obliterated in controls. Recipient immunosuppression facilitated the formation of an integrated ciliated epithelial layer, further demonstrated by the presence of a dense cilia population, a well-developed plasma membrane, and readily recognizable intercellular junctions. Epithelial cellular differentiation markers such as cytokeratin 14 and 18, and cystic fibrosis transmembrane conductance regulator (CFTR) were all positive in allografts under immunosuppression.

CONCLUSIONConcurrent recipient-derived epithelial inoculation with immunosuppression can result in complete re-epithelialization with the recipient phenotype and suppress the luminal obliteration process in heterotopic transplantations.

Allografts ; cytology ; Animals ; Bronchiolitis Obliterans ; surgery ; Epithelial Cells ; cytology ; Female ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Trachea ; cytology ; transplantation ; Transplantation, Heterotopic

Acute respiratory distress syndrome (ARDS) remains a poor prognosis in spite of the recent development of new therapeutic strategies. Cell-based therapy with stem cells has been considered as a promising way for the treatment of vital organ damage. Putative endogenous stem cells have been shown to be located within the adult lung in the basal layer of the upper airways, within or near pulmonary neuroendocrine cell rests, at the bronchoalveolar junction, as well as within the alveolar epithelium. These stem cells are hypothesized to be the source of lung regeneration and repair. But this mechanism seems to be insufficient after lung injury. There is increasing excitement over the last few years with the suggestion that exogenous stem cells may offer new treatment options for ARDS. Exogenous stem cells have the ability to differentiate and function as both airway and lung parenchymal epithelial cells in both in vitro and increasingly in vivo experiments. However, there is great controversy concerning the repair effect of adult stem cells in lung injury. This review evaluates the advances in endogenous respiratory stem cells, and assesses the evidence for the use of stem cells in the repair of lung injury.
Adult Stem Cells ; physiology ; transplantation ; Bone Marrow Transplantation ; Bronchi ; cytology ; Cell Fusion ; Epithelial Cells ; physiology ; Humans ; Pulmonary Alveoli ; cytology ; Respiratory Distress Syndrome, Adult ; therapy

OBJECTIVETo investigate the feasibility of constructing a capsular poly L-lactic acid (PLLA) ureteral stent seeded with autologous urothelial cells using tissue engineering methods.

METHODSThe capsular ureteral stent was constructed by subcutaneously embedding PLLA ureteral stent in the back of beagles for 3 weeks to induce the formation of connective tissue on the surfaces. After decellularization of the stent, the expanded autologous urothelial cells were seeded on the stent. The surface structure and cell adhesion of the stent were observed using HE staining, scanning electron microscope (SEM) and immunocytochemical staining. MTT assay was used to evaluate urothelial cell proliferation on the capsular PLLA ureteral stent and on circumferential small intestinal submucosa graft.

RESULTSHE staining and VIII factor immunohistochemistry revealed numerous capillaries in the connective tissue encapsulating the stent without obvious local inflammatory response. The results of SEM and immunocytochemical staining showed that the capsule contained rich collagenic fibers forming three-dimensional structures, and the seeded autologous urothelial cells could adhere and well aligned on the surface. MTT assay showed normal growth of the cells on the stent as compared with the cells grown on circumferential small intestinal submucosa graft.

CONCLUSIONThe capsular PLLA ureteral stent allows adhesion and proliferation of autologous urothelial cells and shows a potential in applications of constructing tissue-engineered ureter.

Animals ; Bacterial Adhesion ; Cell Proliferation ; Dogs ; Epithelial Cells ; cytology ; transplantation ; Female ; Lactic Acid ; Polyesters ; Polymers ; Stents ; Tissue Engineering ; methods ; Transplantation, Autologous ; Ureter ; surgery ; Urothelium ; cytology

BACKGROUNDHuman amniotic epithelial cells (HAECs), which have several characteristics similar to stem cells, therefore could possibly be used in cell therapy without creating legal or ethical problems. In this study, we transplanted HEACs into the injured spinal cord of rats to investigate if the cells can improve the rats' hindlimb motor function.

METHODSHAECs were obtained from a piece of fresh amnion, labeled with Hoechst33342, and transplanted into the site of complete midthoracic spinal transections in adult rats. The rats (n = 21) were randomly divided into three groups: Sham-operation group (n = 7), cells-graft group (n = 7), and PBS group (n = 7). One rat of each group was killed for histological analysis at the second week after the transplantation. The other six rats of each group were killed for histological analysis after an 8-week behavioral testing. Hindlimb motor function was assessed by using the open-field BBB scoring system. Survival rate of the graft cells was observed at second and eighth weeks after the transplantation. We also detected the myelin sheath fibers around the lesions and the size of the axotomized red nucleus. A one-way ANOVA was used to compare the means among the groups. The significance level was set at P < 0.05.

RESULTSThe graft HAECs survived for a long time (8 weeks) and integrated into the host spinal cord without immune rejection. Compared with the control group, HAECs can promote the regeneration and sprouting of the axons, improve the hindlimb motor function of the rats (BBB score: cells-graft group 9.0 +/- 0.89 vs PBS group 3.7 +/- 1.03, P < 0.01), and inhibit the atrophy of axotomized red nucleus [cells-graft group (526.47 +/- 148.42) microm(2) vs PBS group (473.69 +/- 164.73) microm(2), P < 0.01].

CONCLUSIONTransplantation of HAECs can improve the hindlimb motor function of rats with spinal cord injury.

Amnion ; cytology ; transplantation ; Animals ; Cell Survival ; Epithelial Cells ; transplantation ; Female ; Hindlimb ; physiopathology ; Humans ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; pathology ; physiopathology ; therapy ; Stem Cell Transplantation ; methods

We report our experience with corneal epithelium, grown in vivo, transplantation in three patients with persistent epithelial defect (PED). The three patients had ocular surface disease unresponsive to standard treatments and were therefore chosen for transplantation. They underwent transplantation of epithelial sheets, grown in vivo, to the most affected eye. In vivo cultivation was carried out in the cornea of a living related donor. After epithelialization was completed, the epithelium grown on an amniotic membrane was harvested gently; it was then transplanted into the patient's eye after debridement of fibrovascular tissue. The cultivated epithelium was completely epithelialized by 2 weeks; it was well-differentiated with well-formed hemidesmosome. On immunohistochemical staining, p63, connexin 43, and Integrin beta4 were expressed in the cells on the epithelial sheet. The PED was covered completely and maintained for 4 weeks in all cases. However, corneal erosion recurred after 5 weeks in two cases. This novel technique demonstrates the corneal epithelial cells can be expanded in vivo successfully on denuded amniotic membrane of a healthy cornea and harvested safely. A corneal epithelial sheet, grown in vivo, can be transplanted to treat eye with a severe ocular surface disease, such as total limbal deficiency.
Adult ; Cell Culture Techniques ; Cells, Cultured ; Corneal Diseases/etiology/pathology/*surgery ; Corneal Transplantation/*methods ; Epithelial Cells/cytology/*transplantation ; Epithelium, Corneal/cytology/*transplantation ; Eye Burns/complications ; Humans ; Limbus Corneae/*pathology ; Male ; Middle Aged ; Stem Cells/*pathology ; Stevens-Johnson Syndrome/complications

OBJECTIVETo evaluate the efficacy and risksof the treatment of facial acne scar with dermabrasion combined with Recell® technology.

METHODS30 patents with II or III degree facial acne scars were treated with dermabrasion and Recell® (skin active cell transplantation) technique in our department from October 2010 to October 2011. The affected area in the face was dermabraded with micro motor system. Then a small piece of razor-thickness skin graft was obtained and processed with Recell® kit. Several milliliterof autologous uncultured epidermal cell suspension was applied to the facial wound and covered with appropriate dressings.

RESULTSTheeffectiveness and risks of this treatment was evaluated with regard to wound healing time, postoperative complication rate, erythema period, etc. Wound healing time was shortened to 5-7 days. The erythema period was also observed shortened with this technique. Within the follow up period, no hyperpigmentation was reported in this case serial.

CONCLUSIONDermabrasion combined with Recell® (skin active cell transplantation) technology can provide a safety and effective treatment approach for patients with facial acne scars.

Acne Vulgaris ; complications ; Cell Transplantation ; methods ; Cicatrix ; etiology ; surgery ; Combined Modality Therapy ; methods ; Dermabrasion ; methods ; Epithelial Cells ; transplantation ; Face ; Humans ; Skin ; cytology ; Treatment Outcome ; Wound Healing

OBJECTIVEThe effect of human bone marrow mesenchymal stem cells (hMSCs) on tumor neovascularization were studied.

METHODShMSCs were isolated from human bone marrow by density gradient fractionation and adherence to plastic flasks. hMSCs-EGFP were obtained by pLEGFP-N1 retroviral vector. Flow cytometry was used to detect the cell surface antigen and the differentiation potential of hMSCs-EGFP was investigated under conditioned media. The effect of hMSCs on tumor neovascularization were observed by establishing solid tumor models in BALB/C nude mice. In addition, effect of the conditioned medium used for tumor cells and endothelial cells (EC) cultivation was collected, to detect its effect on the growth and migration rates of hMSC. hMSCs were induced to differentiate into EC in vitro and the migratory effect on HUVEC was also evaluated.

RESULTShMSCs-EGFP, like hMSC, exhibited a fibroblast-like morphological feature, and both had the similar cell surface antigens. They could be induced into osteocytes or adipocytes under the conditioned media. The results not only suggested that hMSCs contributed to tumor neovascularization, but also indicated that most of vessels were host-derived angiogenesis mediated by hMSCs. The mean vascular density (MVD) in suspension group (13.67 ± 1.53) was strikely higher than that in MCF-7 group (5.33 ± 1.42), which showed statistical significance (P < 0.05). Only very few vessels were attributed to hMSCs transdifferentiation into ECs. Tumor cells and ECs can promote hMSCs proliferation and migration through paracrine action. Furthermore, hMSCs were positive for CD31 after 2 weeks induction and HUVEC migration can be facilitated by hMSCs.

CONCLUSIONMSCs have the effect of promoting tumor neovascularization.

Animals ; Bone Marrow Cells ; cytology ; Breast Neoplasms ; pathology ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Epithelial Cells ; cytology ; Humans ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; physiology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microvessels ; pathology ; Neoplasm Transplantation ; Neovascularization, Pathologic ; pathology

BACKGROUNDHuman amniotic epithelial cells (HAECs) are able to secrete biologically active neurotrophins such as brain-derived neurotrophic factor and neurotrophin-3, both of which exhibit trophic activities on dopamine neurons. Previous study showed that when human amniotic epithelial cells were transplanted into the striatum of 6-hydroxydopamine (6-OHDA)-induced Parkinson disease rats, the cells could survive and exert functional effects. The purpose of this study was to investigate the survival and the differentiation of human amniotic epithelial cells after being transplanted into the lateral ventricle of Parkinson's disease (PD) rats, and to investigate the effects of grafts on healing PD in models.

METHODSThe Parkinson's model was made with stereotactic microinjection of 6-hydroxydopamine (6-OHDA) into the striatum of a rat. The PD models were divided into two groups: the HAECs group and the normal saline (NS) group. Some untreated rats were taken as the control. The rotational asymmetry induced by apomorphine of the HAECs group and the NS group were measured post cell transplantation. The expression of nestin and vimentin in grafts were determined by immunohistology. Ten weeks after transplantation the density of tyrosine hydroxylase positive cells in the substantia nigra of the HAECs group, NS group and the untreated group was determined. The differentiation of grafts was determined by TH immunohistology. High performance liquid chromatography (HPLC) was used to determine monoamine neurotransmitter levels in the striatum.

RESULTSThe rotational asymmetry induced by apomorphine of the HAECs group was ameliorated significantly compared to the NS group two weeks after transplantation (P < 0.01). The grafts expressed nestin and vimentin five weeks after transplantation. TH immunohistochemistry indicated that the TH positive cells in the substantia nigra of the HAECs group increased significantly compared to the NS group (P < 0.01). Tyrosine hydroxylase (TH) positive cells in the substantia nigra of the HAEC group and the NS group were decreased compared to the untreated group (P < 0.01). Dopamine and DOPAC levels in the striatum of the HAECs group increased significantly compared to the NS group (P < 0.05). Homovanillic acid (HVA) levels in the striatum of the HAECs group increased significantly compared to the NS group (P < 0.01). In addition dopamine, DOPAC, and HVA levels in the striatum and dopamine levels in the cerebrospinal fluid of the HAECs group and the NS group were decreased compared to the untreated group (P < 0.05).

CONCLUSIONSHuman amniotic epithelial cells could be used to ameliorate the rotational asymmetry induced by apomorphine of the PD models. This could have been due to the increased content of dopamine and its metabolic products, DOPAC and HVA, in the striatum in the PD models.

Amnion ; cytology ; Animals ; Apomorphine ; pharmacology ; Chromatography, High Pressure Liquid ; Epithelial Cells ; cytology ; drug effects ; transplantation ; Female ; Homovanillic Acid ; metabolism ; Humans ; Immunohistochemistry ; Oxidopamine ; toxicity ; Parkinsonian Disorders ; chemically induced ; metabolism ; therapy ; Rats ; Rats, Sprague-Dawley