Objective To study the culture and purification of the fetal mouse liver mesenchymal stem cells(MSCs) in vitro and to investigate their differentiation potential and the composite ability with true bone ceramic(TBC). Methods The single cell suspension of MSCs was primarily cultured and passaged, which was prepared from the fetal mouse liver; the flow cytometry was applied to detectCD29, CD34, CD44 and CD45. The osteogenic differentiation was induced in chemical inducing system; the osteogenic induction potency was tested. The purified fetal mouse liver MSCs were compounded with TBC covered with collagen type Ⅰ in vitro and the cell attachment and proliferation to the TBC were observed. Results The primary MSCs of fetal mouse liver were easy to culture in vitro. They proliferated well and were easy to subcultured. The proliferation ability of primary and passaged MSCs was similar. Flow cytometric analysis showed the positive results for CD29, CD44 and the negative results for CD34, CD45. After 7 days of induction, the MSCs expressed collagen type I and alkaline phosphatase(ALP) highly. After 14 days of induction, the fixed quantity of ALP increased significantly. After 28 days of induction, calcium accumulation was observed by Von Kossa’s staining. Many liver MSCs attached to the surface of TBC. Conclusion The MSCs of the fetalmouse liver can be obtained, subcultured and purified easily. After culturing in chemical inducing system, the MSCs of fetal mouse liver can be successfully induced to osteoblast-like cells, attach to the surface of TBC and proliferate well.
ObjectiveTo systematically review the prognosis of fetal isolated hyper echogenic kidneys (IHEK) on prenatal ultrasound examination. MethodsPubMed, EMbase, Web of Science, WanFang Data, and CNKI databases were electronically searched to collect cross-sectional studies on the prognosis of fetal IHEK on prenatal ultrasound examination from January 1990 to January 2021. Two reviewers independently screened literature, extracted data, and assessed the risk of bias of included studies; then, meta-analysis was performed by using R 3.5.2 software. ResultsA total of 9 cross-sectional studies involving 348 fetuses were included. The results of meta-analysis showed that the incidence rate of live births was 79% (95%CI 69% to 88%), termination of pregnancy/neonatal mortality (TOP/NND) was 30% (95%CI 15% to 45%), normal kidneys was 34% (95%CI 15% to 53%), autosomal recessive polycystic kidney disease (ARPKD) was 21% (95%CI 12% to 30%), autosomal dominant polycystic kidney disease (ADPKD) was 13% (95%CI 5% to 21%), and multicystic dysplastic kidney (MCDK) was 4% (95%CI 2% to 7%). Subgroup analysis showed that the prognosis of normal amniotic fluid subgroup was significantly superior to that of reduced amniotic fluid subgroup. ConclusionCurrent evidence shows that the incidence of adverse pregnancy outcomes in patients with IHEK on prenatal ultrasound examination is high, the prognosis is superior when IHEK with normal amniotic fluid volume, and is worse when with small amniotic fluid volume. Due to limited quality and quantity of the included studies, more high-quality studies are required to verify the above conclusion.
Objective To investigate the feasibility of fetal liver cells for liver tissue engineering, the supporting function of poly L lactic acid (PLLA) scaffold for fetal liver cells and the effects of oncostatin M (OSM), nicotinamide (NA) and dimethyl sulfoxide(DMSO) on growth and hepatic differentiation. Methods After three dimensional PLLA scaffolds having a porous structure were prepared by using NH 4HCO 3 particle, fetal liver cells obtained from E14.5 C57BL/6CrSlc murine embryos were inoculated in the scaffolds. Cells were cultured in Williams’E medium with or without OSM, NA and DMSO for 30 days. Changes in cell number, liver-specific function, and cellular morphology were observed. Results When compared with in monolayer culture, cell number and albumin secretion increased obviously in three-dimensional PLLA. Alburmin secretion increased slightly in OSM group of monolayer culture, but increased obviously in OSM groupo of PLLA culture and in OSM/NA/DMSO group of both monlayer and PLLA cultures. Conclusion The three-dimensional PLLA scaffold is a good supporting material for the cultivation of tetal liver cells. OSM, NA and DMSO remarkaly stimulated maturation of hepatic parenchymal cells in vitro in terms of morphology and liver-specific function.
【摘要】 目的 观察胎羊宫内心脏介入手术胎羊血气及血浆炎性细胞因子的变化。方法 8只怀孕双胎山羊,双胎之一为实验组,在相同麻醉条件下,实验组进行胎羊心脏介入治疗,并抽取血样标本。监测胎羊的心率、血气、乳酸值,运用ELISA法检测治疗组及对照组胎羊白介素(IL)1、IL6、IL8及肿瘤坏死因子(TNFα)。结果 2只胎羊因手术中发生心包填塞死亡,存活的6只胎羊手术前pH值较手术后有明显下降(Plt;005),手术前后乳酸浓度上升(Plt;005),PCO2、PO2差异无统计学意义(Pgt;005),手术前血浆IL1、IL6、IL8的浓度较手术后高(Plt;005),手术前后TNFα的浓度变化无统计学意义(Pgt;005)。结论 胎羊宫内心脏介入手术可引起胎羊血浆pH值下降,乳酸浓度上升,及细胞因子IL1、IL6、IL8浓度上升。【Abstract】 Objective To observe the change of blood gas and inflammatory cytokines during intrauterine cardiac intervention surgery on the fetal lambs. Methods Eight pregnant goats with two fetal in each goat were included. With the same anesthesia condition, one of the twin fetus was chose to perform the intrauterine cardiac intervention surgery. The fetal heart beating rate was monitored, and blood samples of the fetus were taken to do the blood gas analysis and to detect the concentration of inflammatory cytokines (IL1, IL6, IL8, and TNFα). Results Two of the eight fetal lambs which was died in the operation because of pericardial tapenade. In the other six survived fetus, the PH was lower than after the surgery, and the concentrations of lactic acid, IL1, IL6, and IL8 are higher than after the surgery. There was no significant difference of PCO2,PO2 and TNFα between before and after the surgery. Conclusion The intrauterine cardiac intervention surgery can make the PH of fetal plasma lower and the concentrations of lactic acid and IL1, IL6, IL8 higher.
PURPOSES:To investigate the time of neuronie apoptosis in the retinas of Imman fetuses,and its relations with neuronie proliferation and differentiation, METHODS:The retinas of 27 human fetuses from 8th to 38th week of R,~til- ization age and 3 adults were studied by TdT-mediated dUTP nick end labelling(TUNEL) method. RESULTS:Tbe nuctei of labeled apoptotic cells were charaeterised by nuclear marginization,ehromatln condensation and cleseent shape,and some apoptotie bodies were visible in the specimens. The apoptosis of neuroepithelium of fetal rclina took place during 8th to 18th week, Apoptosis of ganglion cells were observed from 1256 to 18th week. The apoptos[s of pholorec, plors were formd from 14th to 2Ist week ,while thai of bipolar neurones and M~ller cells were found from ldth to 28th week. No apoptosb of ocstones were observed in the retinas after 28th week of fertilization age and within the retinas of adults. CONCLUSION:The proliferating cells of neuroepithelium and Ihe neurones which just differetiated from fetal retina might partly undergo apoptosis. The time of apoptosls of differentiated neurones was consistent with the time of the synapses formation between neurones and their targel cells. （Chin J Ocul Fundus Dis,1997,13:67 -69 ）
Objective To investigate the possibility of culturing human oral keratinocyte using autologous serum in order to provide theoretical and technical foundation for clinical application of tissue engineering oral mucosa epithelium.Methods The human oral keratinocytes were cultured by the medium containing different concentrations of autologous serum(10%,20%,30%)and fetalbovine serum (10%), respectively. The growth conditions for the cell and the mucosa epithelium in the groups were observed, the cell growth curves were drawn, and the population doubling time (PDT) was counted. Results The results showed that the human oral keratinocyte could proliferate well in the medium containing autologous serum or fetal bovine serum. The differences in the 24hour clone rate and PDT were not significant. Both the area and the thickness of the cultured oral epithelium increased with the increase of the autologous serum concentration, and the difference between autologous serum and fetal bovine serum was significant, especially with the medium containing 20% autologous serum( P＜0.05) . The human nature of the cultured epithelium was demonstrated by the immunofluorescent mouse anti-HLA antigen. Conclusion The autologous serum can replace the fetal bovine serum to culture the oral keratinocyte well, and the cultured oral mucosa epithelium can be better differentiated in the autologous serum than in the fetal bovine serum.
Abstract In order to repair the bone defect afteroperation of benign lesion of extremity, the fetal demineralized bone was applied in 10 cases. These cases were followed up for 6 months to 8 years. The results showed that the grafted bone was integrated with the host bone in 6 months. Noadverse effect was found. The demineralized bone did not induce rejection. The advantages of using fetal demineralized bone were as follows: easily obtainable,its preparation and method of storage simple, and low finacial cast.
Objective To investigate effects of neural retina on development of the structure of outer blood retinal barrier in embryogenesis. Methods The retinal neural epithelium (RNE) and pigment epithelium (RPE) layers of 150, 120 and 90 embryonic chicken eyes incubated for 7,10, and 14 days were peeled off. RNE was used to prepare the culture medium with different conditions (7drcSF3, 10drcSF3, 14drcSF3). RPE cells of 7- and 14-incubated chicken embryos were cultured on laminin-coated transwell filter. The SF3, 7drcSF3, 10drcSF3 , 14drcSF3 medium were used respectively in the apical chamber and SF2 was used in basolateral chamber. After the formation of monolayer, the transepithelial electrical resistance of the RPE was detected. After the fixation of RPE cells, the condition of the tight junction among the cells was observed by immunohis tochemistry and transmission electron microscopy. Results For the RPE cells of 7-and 14-day incubated embryonic eyes, the difference of TER in various medium of SF3/SF2, 7drcSF3/SF2, 10drcSF3/SF2, 14drcSF3/SF2 was statistically significant (P＜0.01). The polarity of RPE cells was induced and the netlike tight junctional strands was urged in the retina-conditioned medium. Conclusion The neural retina may actively promote the formation of the structure of outer blood retinal barrier. (Chin J Ocul Fundus Dis，2004，20：237-240)
OBJECTIVE: To explore the effects of different methods of fetal spinal cord(FSC) tissue transplanted on reversing the axotomy-induced neurons atrophy of adult rats injured spinal cord. METHODS: One hundred and twenty adult rats received lumbar spinal cord hemisection. Experimental rats were divided into five groups, the control group(Group A); spinal cord hemisection only(Group B); spinal cord hemisection plus FSC transplant (Group C); spinal cord hemisection plus FSC transplant plus pedicled paraspinal muscle(Group D); spinal cord hemisection plus FSC transplant plus pedicled omentum (Group E). Combined behavioral scores(CBS), somatosensory evoked potentials (SEP), motor evoked potentials(MEP) were examined to evaluate the recovery of neurological function after operation. Rats were sacrificed after 1, 4 and 12 weeks. Nissl stained section was used for neurons quantitative image analysis. The positive cells were quantitative analysis by computer image analysis system. RESULTS: The different methods of FSC tissue transplantation could prevent the neurons atrophy secondary to axon injury of spinal cord in adult rats. The size of neurons were observed in five groups, they were group E gt; group D gt; group C gt; group B gt; group A (P lt; 0.05). Those increases in size of neurons were paralleled with a significant improvement in neurological function recovery. CONCLUSION: It indicates that the different methods of FSC tissue transplantation can maintain the neurons morphology and improve the neurological function of rats.
Objective To observe the effect of BMSCs transplantation on gene and protein expression of VEGF receptor fetal l iver kinase 1 (Flk-1) after spinal cord injury (SCI), and to investigate the mechanism of repairing the SCI by BMSCs transplantation. Methods BMSCs were isolated and cultured from five 4-week-old male Wistar rats weighing100-120 g. The SCI model was made by using the modified Allen’s impactor device. Eighty-one adult female Wistar rats weighing 220-250 g were randomly divided into 3 groups: sham-operated group (group A, n=21), in which spinous process and vertebral plate of thorax 8-10 spinal cord segment were removed; DMEM group (group B, n=30), in which rats received four injections of DMEM in the peri-lesion area; and BMSCs group (group C, n=30), in which rats received four injections of BMSCs in the peri-lesion area. The changes of Flk-1 mRNA expression in rats’ spinal cord tissues were detected with RT-PCR method 1, 3 and 5 days after transplantation. The expression of Flk-1 protein was observed by using immunohistochemical technology in spinal cord 3, 7 and 14 days after transplantation. Results Morphology of the primary cultured BMSCs was various. Cell morphology tended to be uniform with the accumulation of passages, which appeared flat and spindle-shaped. RT-PCR results showed that there was no significant differences (P gt; 0.05) in Flk-1 mRNA expression between group C and group B at different time points after transplantation. But Flk-1 mRNA levels of group B and group C significantly increased and peaked 1 day after transplantation (P lt; 0.01), and then decreased 3 days after transplantation (P lt; 0.01) compared with that of group A, and were still higher than that of group A 5 days after transplantation (P lt; 0.05). Immunohistochemical staining results revealed that the expression of Flk-1 in group B was enhanced 3 and 7 days after transplantation compared with group A, which was significantly different (P lt; 0.01). There was no significant difference in the expression of Flk-1 between group B and groupp A 14 days after transplantation (P gt; 0.05). There was no significant difference in Flk-1 protein expression between group C and group B 3 days after transplantation (P gt; 0.05). The expression of Flk-1 protein in group C was significantly higher than that in group B 7 and14 days after transplantation (P lt; 0.01). Conclusion BMSCs transplantation after SCI does not have regulatary effect onthe expression of Flk-1 mRNA, but it does upregulate the Flk-1 protein expression, which may be one of the mechanisms of repairing SCI.