Résumé
OBJECTIVE@#To explore the effect of hypoxia-supported umbilical cord mesenchymal stem cell (UC-MSC) on the expansion of cord blood mononuclear cell (MNC) in vitro.@*METHODS@#The isolated cord blood mononuclear cells were inoculated on the preestablished umbilical cord mesenchymal stem cell layer and cultured under hypoxic conditions (3% O2) and the experimental groups were normoxia (MNCs were cultured under normoxic conditions), hypoxia (MNCs were cultured under hypoxic conditions), UC-MSC (MNCs were cultured with UC-MSC under normoxic conditions), and UC-MSC+hypoxia (MNCs were cultured with UC-MSC under hypoxic conditions). To further investigate the combinational effect of 3 factors of SCF+FL+TPO (SFT) on expansion of cord blood MNCs in vitro in hypoxia-supported UC-MSC culture system, the experiments were further divided into group A (MNCs were cultured with UC-MSC and SFT under normoxic conditions), group B (MNCs were cultured with UC-MSC under hypoxic conditions), group C (MNCs were cultured with UC-MSC and SFT under hypoxic conditions). The number of nucleated cells (TNC), CD34+ cell, CFU and CD34+CXCR4+, CD34+CD49d+, CD34+CD62L+ cells of each groups were detected at 0, 7, 10 and 14 days, respectively.@*RESULTS@#Compared with group hypoxia and UC-MSC, group UC-MSC+hypoxia effectively promoted the expansion of TNC, CD34+ cell and CFU, and upregulated the expression level of adhesion molecule and CxCR4 of the cord blood CD34+ cell(P<0.05). After culturing for 14 days, compared with group A and group B, group C effectively promoted the expansion of cord blood MNC at different time points(P<0.05), and the effect of group A was better than that of group B at 7 and 10 days(P<0.05).@*CONCLUSION@#Hypoxia-supported UC-MSC efficiently promoted the expansion and expression of adhesion molecule and CXCR4 of cord blood CD34+ cell, and the effect of expansion could be enhanced when SFT 3 factors were added.
Sujets)
Humains , Cellules cultivées , Sang foetal , Prolifération cellulaire , Cordon ombilical/métabolisme , Cellules souches mésenchymateuses , Antigènes CD34/métabolisme , Hypoxie/métabolismeRésumé
Impaired immunomodulatory capacity and oxidative stress are the key factors limiting the effectiveness of mesenchymal stem cell transplantation therapy. The present study was aimed to investigate the effects of jujuboside A (JuA) on the protective effect and immunomodulatory capacity of human umbilical cord mesenchymal stem cells (hUC-MSCs). Hydrogen peroxide was used to establish an oxidative damage model of hUC-MSCs, while PBMCs isolated from rats were used to evaluate the effect of JuA pre-treatment on the immunomodulatory capacity of hUC-MSCs. Furthermore, Hoechst 33258 staining, lactate dehydrogenase test, measurement of malondialdehyde, Western blot, high-performance liquid chromatography; and flow cytometry were performed. Our results indicated that JuA (25 μmol·L-1) promoted the proliferation of hUC-MSCs, but did not affect the differentiating capability of these cells. JuA pre-treatment inhibited apoptosis, prevented oxidative damage, and up-regulated the protein expression of nuclear factor-erythroid factor 2-related factor 2 and heme oxygenase 1 in hUC-MSCs in which oxidative stress was induced with H2O2. In addition, JuA pre-treatment enhanced the inhibitory effect of hUC-MSCs against abnormally activated PBMCs, which was related to stimulation of the expression and activity of indoleamine 2,3-dioxygenase. In conclusion, our results demonstrate that JuA pre-treatment can enhance the survival and immunomodulatory ability through pathways related to oxidative stress, providing a new option for the improvement of hUC-MSCs in the clinical setting.
Sujets)
Animaux , Humains , Rats , Différenciation cellulaire , Peroxyde d'hydrogène/métabolisme , Cellules souches mésenchymateuses , Stress oxydatif , Saponines , Cordon ombilical/métabolismeRésumé
The purpose of this study is to examine the changes in the umbilical cord in women diagnosed with gestational diabetes mellitus In this study, as a control group human placental tissues from normotensive pregnancies was collected from diabetic women at 2835 weeks of gestation. Gestational diabetes (n= 20) and normal umbilical cord (n= 20) for a total of 40 units were received.GDM groups compared to the control group was significantly higher values was detected (p<0.01). In GDM group, light microscopy showed erosion of the endothelium and complete rupture of theumbilicalvessels resulting in extravasation of blood within Wharton's jelly. it was observed that the cytoplasmic fragments and cell infiltration of the spill to the subepithelial layer of apoptotic cell PECAM-1 positive reaction showed. E-Cadherin in endothelial side surface of diabetes group showed weak expression in the nucleus and showed positive reaction in smooth muscle.
El objetivo fue examinar los cambios que presenta el cordón umbilical de mujeres con diagnóstico de diabetes mellitus gestacional (DMG). Se incluyeron en el grupo control muestras de tejidos placentarios humanos de embarazos normotensos y de mujeres diabéticas de entre 2835 semanas de gestación. Las muestras se divieron en cordones umbilicales con cambios de DMG (n= 20) y cordones umbilicales normales (n= 20), constituyendo un total de 40 muestras. El grupo de DMG, en comparación con el grupo control, presentó valores significativamente más elevados (p<0,01). En el grupo de DMG, la microscopía óptica demostró la erosión del endotelio y la ruptura completa de los vasos umbilicales, resultando en la extravasación de sangre dentro de la gelatina . Se observaron fragmentos citoplasmáticos e infiltración celular de la capa subepitelial de células apoptóticas mostró una reacción positiva a PECAM-1. En el grupo de DMG, la E-cadherina de la superficie lateral endotelial mostró una expresión débil en el núcleo y una reacción positiva en el músculo liso.
Sujets)
Humains , Femelle , Grossesse , Cadhérines/métabolisme , Diabète gestationnel , Antigènes CD31/métabolisme , Cordon ombilical/métabolisme , Cordon ombilical/ultrastructure , Immunohistochimie , MicroscopieRésumé
Canine mesenchymal cells (MSCs) derived from Wharton's jelly were co-cultured, then supplemented or not supplemented with platelet rich plasma (PRP) and demineralized bone matrix (DBM) to verify osteogenic differentiation. Osteoblastic differentiation followed by mineralized bone matrix production was found to be significantly higher (p < 0.05) when MSCs were associated with PRP/DBM in culture after 14-21-days of induction. Osteopontin and osteocalcin gene expression were significantly superior (p < 0.05) under the same culture conditions after 21 days of observation. In conclusion, addition of PRP to DBM co-cultured with MSCs successfully induced osteogenesis in vitro.
Sujets)
Animaux , Chiens , Technique de déminéralisation de l'os/médecine vétérinaire , Trame osseuse/métabolisme , Différenciation cellulaire , Cellules cultivées , Techniques de coculture/médecine vétérinaire , Cellules souches mésenchymateuses/métabolisme , Ostéogenèse , Plasma riche en plaquettes/métabolisme , Cordon ombilical/métabolismeRésumé
A importância do estudo do sangue do cordão umbilical (SCU) tem sido verificada pela presença de células-tronco hematopoéticas no SCU humano. O modelo experimental em cão tem propiciado informações importantes nos transplantes em humanos. Entretanto, apesar da importância do SCU e do modelo experimental em cães, não existem estudos sobre a eletroforese de proteínas séricas do cordão umbilical canino. No presente protocolo experimental, foi feita a colheita de SCU de 20 neonatos caninos, o qual foi utilizado para o fracionamento eletroforético de suas proteínas. Os valores médios absolutos obtidos para proteínas séricas totais, albumina, alfa, beta e gamaglobulinas no sangue do cordão umbilical de cães ao nascimento foram 3,09±0,59; 1,51±0,38; 0,87±0,17; 0,68±0,12 e 0,03±0,01, respectivamente. Todos os resultados apresentaram-se abaixo dos níveis reportados para animais adultos, devido à passagem das proteínas e suas frações ocorrerem principalmente através do colostro e pela imaturidade hepática. Em todos os traçados eletroforéticos do SCU de cães, foi observada uma pequena curva entre alfaglobulina 2 e betaglobulina 1, não relatada no soro de cães adultos. Portanto, neste experimento, foram observadas diferenças quantitativas no traçado eletroforético das proteínas séricas do SCU de cães, ao nascimento, diferindo-os, neste particular, de cães em diferentes fases da vida pós-colostral.
Sujets)
Sang , Cordon ombilical/cytologie , Cordon ombilical/métabolisme , Chiens , ÉlectrophorèseRésumé
Fez-se cuidadosa revisäo bibliográfica, procurando mostrar a importância do estudo histoquímico da placenta humana nas diversas idades gestacionais. Vislumbram-se novos passos que poderäo ser dados quando do funcionamento do Laboratório de Histoenzimologia, a qual será instalado em futuro próximo no Departamento de Patologia da Faculdade de Medicina da Universidade Federal do Ceará