艾灸对快速老化模型小鼠海马神经干细胞分化的影响

目的:研究艾灸疗法对快速老化模型(SAMP8)小鼠海马神经干细胞(NSCs)分化的影响。方法:24只SAMP8小鼠随机分为模型组与艾灸组,12只抗快速老化型(SAMR1)小鼠作为正常对照组,艾灸组选用"百会"穴进行艾灸治疗。每天治疗1次,7d为1个疗程,共治疗3个疗程。处死前1周开始给予小鼠50mg/kg的溴脱氧尿嘧啶核苷(BrdU)腹腔注射,治疗结束后,取海马组织,用免疫荧光双标方法检测NSCs分化。结果:①各组小鼠海马均有新生神经元免疫荧光双标阳性细胞表达。与模型组比较,艾灸组有促进或诱导NSCs向成熟神经元及未成熟神经元分化的倾向(P0.05)。②模型组成熟星形胶质细胞神经胶质纤维酸性蛋白表达增多,艾灸能使其表达下降(P0.05);模型组未成熟星形胶质细胞表达减少,艾灸能促进其表达(P0.05)。③与对照组比较,模型组少突胶质细胞增多(P0.05);艾灸能减少其表达(P0.05)。结论:经过3个疗程的艾灸治疗,能抑制小鼠海马NSCs向少突胶质细胞分化,促进其向未成熟星形胶质分化,同时其也有向神经元分化倾向。     

Effects of chronic cigarette smoke inhalation on the development of senile lung in senescence-accelerated mouse

The senescence-accelerated mouse (SAM) manifests most of the features of function and morphology in the senile lung with aging. However, little is known about the effects of age and cigarette smoke on alterations of the lung in SAM. In the present study, we examined the effects of chronic cigarette smoke inhalation and age on the function and morphology of lungs in two strains of SAM, SAMP2 (senescence-prone strain) and SAMR1 (senescence-resistant strain), from 6 months of age (young) and 18 months of age (aged). After 4 weeks of cigarette smoke inhalation, a small but significant airspace along with a leftward shift of the pressure-volume (P-V) curve was observed in young SAMP2, but not in SAMR1. However, the airspace size of young SAMP2 with cigarette inhalation was smaller than that in aged SAM with air inhalation, suggesting that the effect of age may be greater than that of the small burder of tobacco smoke on the lung alterations in SAMP2. In the aged SAM, there were no differences in function and structure between tobacco-exposed and air-exposed mice. Because the changes in the lungs of young SAMP2 exposed to cigarette smoke were partly simulated with age-related alterations in human lung, and because age-dependent changes of lungs were clearly investigated in SAMP2, this strain may be an interesting animal model for investigating the effects of age and/or cigarette smoke on alterations in lung structure and function.     

The effect of age on osteogenic,adipogenic and proliferative potential of female adipose‐derived stem cells

Human adipose tissue is an ideal source of autologous cells that is both plentiful and easily obtainable in large quantities through the simple surgical procedure of liposuction. The stromal vascular fraction of adipose tissue contains a stem cell population, adipose‐derived stem cells (ASCs), capable of adipogenic, osteogenic, myogenic and chondrogenic differentiation. These cells have already been recognized to possess great therapeutic potential in tissue engineering and regeneration. In this study, we sought to determine the effect of donor age on the growth kinetics and differentiation potential of ASCs. For this, ASCs were isolated from liposuctioned adipose tissue obtained from female patients in the age range 20–58 years. Population doubling time was calculated over 2 weeks and differentiation potential was determined by assaying for adipogenesis and osteogenesis. ASCs obtained from older donors appeared to have a slower rate of proliferation, but this relationship was not significant. While adipogenic potential was unrelated to donor age, a distinct relationship between donor age and osteogenic potential was observed. The aetiology of this age‐dependent change in osteogenic potential was not due to any changes in the number of precursors with osteogenic capacity in the adipose sample. These findings have important implications for emerging cell‐based therapeutic strategies, such as tissue engineering, in addition to treatment of various metabolic bone disorders including osteoporosis. Copyright © 2009 John Wiley & Sons, Ltd.     

Molecular‐ and microarray‐based analysis of diversity among resting and osteogenically induced porcine mesenchymal stromal cells of several tissue origin

Mesenchymal stromal cells (MSCs) play a pivotal role in modern therapeutic approaches in bone‐healing disorders. Although bone marrow‐derived MSCs are most frequently used, the knowledge that many other adult tissues represent promising sources for potent MSCs has gained acceptance. In the present study, the osteogenic differentiation potential of porcine skin fibroblasts (FBs), as well as bone marrow‐ (BMSCs), adipose tissue‐ (ASCs) and dental pulp‐derived stromal cells (DSCs) were evaluated. However, additional application of BMP‐2 significantly elevated the delayed osteogenic differentiation capacity of ASC and FB cultures, and in DSC cultures the supplementation of platelet‐rich plasma increased osteogenic differentiation potential to a comparable level of the good differentiable BMSCs. Furthermore, microarray gene expression performed in an exemplary manner for ASCs and BMSCs revealed that ASCs and BMSCs use different gene expression patterns for osteogenic differentiation under standard media conditions, as diverse MSCs are imprinted dependent from their tissue niche. However, after increasing the differentiation potential of ASCs to a comparable level as shown in BMSCs, a small subset of identical key molecules was used to differentiate in the osteogenic lineage. Until now, the importance of identified genes seems to be underestimated for osteogenic differentiation. Apparently, the regulation of transmembrane protein 229A, interleukin‐33 and the fibroblast growth factor receptor‐2 in the early phase of osteogenic differentiation is needed for optimum results. Based on these results, bone regeneration strategies of MSCs have to be adjusted, and in vivo studies on the osteogenic capacities of the different types of MCSs are warranted. Copyright © 2016 The Authors Tissue Engineering and Regenerative Medicine published by John Wiley & Sons, Ltd.     

Osteogenic potential of rat stromal cells derived from periodontal ligament

Various mesenchymal stromal cells (MSCs) have been applied to regenerative medicine. MSCs derived from periodontal tissue could also be a useful cell source for alveolar bone regeneration. However, only a few attempts of direct comparisons have been made between MSCs from periodontal tissues and those from other somatic tissues. The purpose of this study was to clarify the osteogenic characteristics of mesenchymal stromal cells derived from bone marrow (BMSCs), adipose tissue (ASCs) and periodontal ligament (PDLSCs). BMSCs, ASCs and PDLSCs were isolated from Fisher 344 rats. After 1 week of primary culture, stromal cells were subjected to cell surface analysis and osteogenic differentiation. The cells were subcultured for 2 weeks with and without osteogenic supplements (OS), followed by biochemical and histological analyses. With regard to cell surface antigens, all MSCs were positive for CD29 and CD90 and negative for CD45. With regard to osteogenic differentiation, BMSCs with OS had the highest ALP activity, calcium uptake and osteocalcin content. Without OS, PDLSCs had the highest levels of these bone differentiation markers. RT-PCR analysis and histological analysis showed similar trends. These results indicate that PDLSCs are an ideal candidate for alveolar bone regeneration.     

Ectopic bone formation by aggregated mesenchymal stem cells from bone marrow and adipose tissue: A comparative study

Tissue engineered constructs (TECs) based on spheroids of bone marrow mesenchymal stromal cells (BM‐MSCs) combined with calcium phosphate microparticles and enveloped in a platelet‐rich plasma hydrogel showed that aggregation of MSCs improves their ectopic bone formation potential. The stromal vascular fraction (SVF) and adipose‐derived MSCs (ASCs) have been recognized as an interesting MSC source for bone tissue engineering, but their ectopic bone formation is limited. We investigated whether aggregation of ASCs could similarly improve ectopic bone formation by ASCs and SVF cells. The formation of aggregates with BM‐MSCs, ASCs and SVF cells was carried out and gene expression was analysed for osteogenic, chondrogenic and vasculogenic genes in vitro. Ectopic bone formation was evaluated after implantation of TECs in immunodeficient mice with six conditions: TECs with ASCs, TECs with BM‐MSC, TECs with SVF cells (with and without rhBMP2), no cells and no cells with rhBMP2. BM‐MSCs showed consistent compact spheroid formation, ASCs to a lesser extent and SVF showed poor spheroid formation. Aggregation of ASCs induced a significant upregulation of the expression of osteogenic markers like alkaline phosphatase and collagen type I, as compared with un‐aggregated ASCs. In vivo, ASC and SVF cells both generated ectopic bone in the absence of added morphogenetic proteins. The highest incidence of bone formation was seen with BM‐MSCs (7/9) followed by SVF + rhBMP2 (4/9) and no cells + rhBMP2 (2/9). Aggregation can improve ectopic bone tissue formation by adipose‐derived cells, but is less efficient than rhBMP2. A combination of both factors should now be tested to investigate an additive effect.     

Linkage of decreased bone mass with impaired osteoblastogenesis in a murine model of accelerated senescence.

Bone marrow is the principal site for osteoclastogenesis and osteoblastogenesis; and an increase in the former has been linked with bone loss caused by acute loss of gonadal steroids. We have now used an established murine model of accelerated senescence and osteopenia (SAMP6) to test the hypothesis that reduced osteoblastogenesis is linked with decreased bone mass. At 1 mo of age, the number of osteoblast progenitors in SAMP6 marrow was indistinguishable from controls; however a threefold decrease was found at 3-4 mo of age. Impaired osteoblast formation was temporally associated with decreased bone formation and decreased bone mineral density, as determined by histomorphometric analysis of tetracycline-labeled cancellous bone and dual-energy x-ray absorptiometry, respectively. Osteoclastogenesis determined in ex vivo bone marrow cultures was also decreased in these mice, as was the number of osteoclasts in histologic sections. Moreover, unlike controls, senescence-accelerated mice failed to increase osteoclast development after gonadectomy. The osteoclastogenesis defeat was secondary to impaired osteoblast formation as evidenced by the fact that osteoclastogenesis could be restored by addition of osteoblastic cells from normal mice. These findings provide the first demonstration of a link between low bone mineral density and decreased osteoblastogenesis in the bone marrow and validate the senescence-accelerated mouse as a model of involutional osteopenia.     

Osteogenic differentiation of two distinct subpopulations of human adipose-derived stem cells: an in vitro and in vivo study

The first stem cells considered for the reconstruction of bone were bone marrow mesenchymal stem cells (BMSCs). Subsequently, cells with similar marker expression panel and differentiation potential were found in new sources of cells, such as adipose tissue. This source of stem cells has a promising future in tissue-engineering applications, considering the abundance of this tissue in the human body, the easy harvesting and the high number of stem cells that are available from such a small amount of tissue. The isolation of the adipose stem cells is generally performed by means of enzymatic digestion of the tissues, followed by a natural selection of the stem cells based on their capacity to adhere to the culture flasks, leading to a quite heterogeneous population. This constitutes a major drawback for the use of these cells, since the heterogeneity of the cell culture obtained can compromise their proliferation and differentiation potential. In the present study we have analysed the in vitro and in vivo behaviour of two selected subpopulations with high osteogenic potential. For this purpose, ASCs(CD29+) and ASCs (STRO-1+)subpopulations were isolated and in vitro cultured onto a biodegradable polymeric scaffold, using osteogenic medium, before implantation in a nude mice model. The biodegradable polymeric scaffold used is a fibre-mesh structure based on a blend of starch and polycaprolatone (SPCL) that has been successfully used in several bone tissue-engineering studies. The implanted ASCs-scaffold constructs promoted the formation of new bone tissue in nude mice. However, the results obtained show differences in the behaviour of the two ASCs subpopulations under study, particularly regarding their potential to differentiate into the osteogenic lineage, and allowed the indentification of ASCs (STRO-1+) as the best subpopulation for bone tissue-engineering applications.     

骨髓基质干细胞治疗骨不连的新进展

背景：骨髓基质干细胞体外培养增殖力强、易于向成骨细胞及软骨细胞方向分化且成骨性能稳定等特点,成为骨组织工程中合适的种子细胞.目的：总结分析采用骨髓基质干细胞作为种子细胞,分析其直接移植于骨不连部位或复合支架或转基因治疗骨不连所具有的优劣势.方法：检索1992/2011西文生物医学期刊文献数据及CNKI 数据库有关骨不连研究,骨髓基质干细胞分离、培养,在骨不连方面的应用,骨组织工程细胞支架方面的文献,英文检索词为＂bone marrow stromal stem cells,nonunions,repairing,tissue engineering＂,中文检索词为＂骨髓基质干细胞,骨修复,骨不连,组织工程＂.排除重复性研究,保留23篇进一步归纳总结.结果与结论：利用骨髓基质干细胞作为种子细胞,直接植入骨不连部位,或与适当的支架材料结合,或用骨髓基质干细胞作为靶细胞,导入外源目的基因诱导成骨的基因治疗来修复骨缺损的方法,给骨缺损的治疗带来光明的前景.但同时也存在骨髓基质干细胞增殖、分化合适条件难以准确确定,经皮移植自体骨髓基质干细胞植入体内后容易流失,不能在植入部位形成有效的细胞浓度,支架材料尚不能完全符合临床要求,以及如何将骨组织工程与基因治疗的方法结合起来等问题,需要进一步的研究.     

Industrial approach in developing an advanced therapy product for bone repair

Mesenchymal stem cells (MSCs) are multipotent cells with therapeutic applications. The aim of our work was to develop an advanced therapy product for bone repair, associating autologous human adipose‐derived MSCs (ASCs) with human bone allograft (TBF; Phoenix^®). We drew up specifications that studied: (a) the influence of tissue collection procedures (elective liposuction or non‐invasive resection) and patient age on cell number and function; (b) monolayer cell culture conditions and osteodifferentiation and particularly the possibility of reducing stages of culture; and (c) the bone construct preparation and especially the comparison between two types of cells seeded on bone allograft (number of cultured processed lipoaspirate (PLA) cells and monolayer‐expanded ASCs) and cultured for 1, 2 and 3 weeks. The results showed that tissue harvesting techniques and patient age did not affect PLA cell number and ASC cloning efficiency. PLA cells can be directly osteodifferentiated (instead of culturing them in expansion medium first and then differentiating them) and these cells were able to mineralize when they were cultured in an osteogenic medium containing calcium chloride. PLA cells directly seeded on bone allograft for a minimum of 3 weeks of culture in this osteogenic medium expressed osteocalcin and colonized the matrix better than monolayer‐expanded ASCs. This work detailed the specifications of a pharmaceutical laboratory to develop an advanced therapy product and this current approach is promising for bone repair. Copyright © 2009 John Wiley & Sons, Ltd.     

人与家兔脂肪来源干细胞体外培养特性的比较

背景：不同种属来源的脂肪来源干细胞在体外培养时特性是否存在差异目前尚未定论。目的：观察在相同培养条件下,人与家兔脂肪来源干细胞体外培养特性的异同。方法：体外分离人腹部取皮植皮术来源的脂肪来源干细胞、家兔背部皮下脂肪来源的脂肪来源干细胞,体外培养并传代,观察各自生长形态,取第3代脂肪来源干细胞,比较二者生长及增殖能力、表面CD分子鉴定情况及成脂、成骨分化能力。结果与结论：人和家兔皮下脂肪均能在体外分离出＂成纤维细胞样＂贴壁生长呈长梭形的细胞;人脂肪来源干细胞一般6～8d可传代,兔脂肪来源干细胞则需要四五天传代。四唑盐结果显示兔、人脂肪来源干细胞分别在第4,6天达到生长高峰。表面标记流式鉴定二者均显示CD29＋CD31-。体外分离培养的人脂肪来源干细胞和兔脂肪来源干细胞均具有干细胞的培养特性。与人脂肪来源干细胞相比,兔脂肪来源干细胞具有更强的增殖和诱导成脂能力,但诱导成骨能力较差,家兔是做脂肪移植研究实验动物不错的选择。     

Osteogenic potential of bone marrow stromal cells

Bone marrow is the point of origin of several cell types, including stromal cells. Adherent bone marrow stromal cells can differentiate into chondrocytes, adipocytes and osteoblasts. Several substances which modulate the dynamics and differentiation of bone marrow stromal cells have been identified. Recently it has been discovered that those bone marrow cells which are non-adherent in tissue culture for 3 days have osteogenic potential comparable to that of whole bone marrow cells. In the future, implantation of non-adherent bone marrow stromal cells may be of use as an aid in bone fracture healing, similar to whole marrow or adherent stromal cell grafting at present.     

Streamlining the generation of an osteogenic graft by 3D culture of unprocessed bone marrow on ceramic scaffolds

Mesenchymal stromal cells are present in very low numbers in the bone marrow, necessitating their selective expansion on tissue culture plastic prior to their use in tissue-engineering applications. MSC expansion is laborious, time consuming, unphysiological and not economical, thus calling for automated bioreactor-based strategies. We and others have shown that osteogenic grafts can be cultured in bioreactors by seeding either 2D-expanded cells or by direct seeding of the mononuclear fraction of bone marrow. To further streamline this protocol, we assessed in this study the possibility of seeding the cells onto porous calcium phosphate ceramics directly from unprocessed bone marrow. Using predetermined volumes of bone marrow from multiple human donors with different nucleated cell counts, we were able to grow a confluent cell sheath on the scaffold surface in 3 weeks. Cells of stromal, endothelial and haematopoietic origin were detected, in contrast to grafts grown from 2D expanded cells, where only stromal cells could be seen. Upon implantation in nude mice, similar quantities of bone tissue were generated as compared to that obtained by using the conventional number of culture expanded cells from the same donor. We conclude that human osteogenic grafts can be efficiently prepared by direct seeding of cells from unprocessed bone marrow.     

Comparison of infrapatellar and subcutaneous adipose tissue stromal vascular fraction and stromal/stem cells in osteoarthritic subjects

Since inflammatory mechanisms have been postulated to link obesity to osteoarthritis, the current study evaluated the ratio of immune cells to multipotent stromal cells within the infrapatellar fat pad (IPFP) and subcutaneous adipose tissue (SQ) of the knee; each depot has potential as a source of regenerative cells. The immunophenotypes of stromal vascular fraction (SVF) and adipose‐derived stem cells (ASCs) of the IPFP and SQ were determined in tissues from osteoarthritic subjects (n = 7) undergoing total knee replacement. Based on a subset of surface antigens, the immunophenotype of ASCs from SQ of OA subjects was not significantly different from that of relatively healthy and leaner subjects undergoing elective liposuction surgery. Flow‐cytometry comparison of SVF cell populations in the IPFP of OA subjects resembled those within the subject's own matched SQ, with the exception of the endothelial marker CD31⁺, which was significantly greater in cells from SQ. In the OA subjects, lower numbers of capillary‐like structures and higher numbers of stromal and alkaline phosphatase colony‐forming units in the IPFP vs SQ were consistent with this finding; however, ASCs from both depots in OA subjects exhibited comparable adipogenic and osteogenic differentiation potential. Thus, the IPFP contains an ASC and immune cell population similar to that of donor‐matched SQ, making it an alternative ASC source for tissue regeneration. Further studies will be needed to determine whether IPFP immune cell infiltrates play an aetiological role in osteoarthritis equivalent to that shown in diabetes associated with obesity. Copyright © 2012 John Wiley & Sons, Ltd.     

不同幅度持续张应力干预下骨髓基质干细胞的成骨分化

背景：研究证实力学刺激是影响骨改建的重要因素,可促进骨髓基质干细胞骨向分化;但不同幅度力学刺激对骨髓基质干细胞分化的影响尚不明确。目的：观察持续张应力对大鼠骨髓基质干细胞成骨分化的影响。方法：全血贴壁培养法秋取大鼠骨髓基质干细胞。采用Flexercell-4000细胞体外应力加载系统对骨髓基质干细胞施加5％,10％,15％幅度的持续张应力,对照组则不加力培养,频率1Hz,持续时问48h。分刖在加力后1,6,12,24,48h检测成骨标记物碱性磷酸酶、I型胶原、骨钙素mRNA及成骨特异性转录因子Runx2的mRNA及蛋白表达。结果与结论：5％和10％持续张应力作用下,骨髓基质干细胞的成骨标记基因碱性磷酸酶、I型胶原、骨钙素mRNA的表达较对照组升高（P〈0．05）,10％张力组升高的时间均较5％张力组早、幅度较高。15％持续在加力6h时可促进骨髓牲质干细胞碱性磷酸嗨、I魁胶原mRNA的表达（P〈0．05）、随后表达下降,加力48h后上述指标均低于对照组（P〈0．05）,骨钙素mRNA的表达加力6h后均低于对照组（P〈O．05）。5％张力组仅加力24h后骨髓基质干细胞Run×2蛋白表达高于对照组（P〈0．05）,10％,15％张力组加力6h后Run×2蛋白表达均高于对照组（P〈0．05）。结果证实,5％,10％,15％持续张应力均可更有效地促进骨髓基质干细胞的骨向分化,10％持续张力的促进效应更显著。     

Short‐term evaluation of electromagnetic field pretreatment of adipose‐derived stem cells to improve bone healing

An electromagnetic field is an effective stimulation tool because it promotes bone defect healing, albeit in an unknown way. Although electromagnetic fields are used for treatment after surgery, many patients prefer cell‐based tissue regeneration procedures that do not require daily treatments. This study addressed the effects of an electromagnetic field on adipose‐derived stem cells (ASCs) to investigate the feasibility of pretreatment to accelerate bone regeneration. After identifying a uniform electromagnetic field inside a solenoid coil, we observed that a 45 Hz electromagnetic field induced osteogenic marker expression via bone morphogenetic protein, transforming growth factor β, and Wnt signalling pathways based on microarray analyses. This electromagnetic field increased osteogenic gene expression, alkaline phosphate activity and nodule formation in vitro within 2 weeks, indicating that this pretreatment may provide osteogenic potential to ASCs on three‐dimensional (3D) ceramic scaffolds. This pretreatment effect of an electromagnetic field resulted in significantly better bone regeneration in a mouse calvarial defect model over 4 weeks compared to that in the untreated group. This short‐term evaluation showed that the electromagnetic field pretreatment may be a future therapeutic option for bone defect treatment. Copyright © 2012 John Wiley & Sons, Ltd.     

Modulation of infection-induced inflammation and locomotive deficit and longevity in senescence-accelerated mice-prone (SAMP8) model by the oligomerized polyphenol Oligonol.

Oligonol is produced from the oligomerization of polyphenols (typically proanthocyanidin from a variety of fruits such as lychees, grapes, apples, persimmons, etc.) and contains catechin-type monomers and oligomers of proanthocyanidins. The ability of Oligonol to affect infection-dependent eye inflammation, locomotion and longevity in senescence-accelerated prone mice (SAMP8) (a model of senescence acceleration and geriatric disorders with increased oxidative stress and neuronal deficit) was investigated. Oligonol (60mg/kg) significantly modulated the extent of inflammation scores in the eye of SAMP8 mice. Examination of the mice indicated infection with mouse hepatitis virus and pinworm (Syphacia obvelata) in both males and females and with the intestinal protozoa (trichomonad) in males. A comparison of the two groups (using log-rank test) and the difference in the mean life span between groups (using Student's t-test) indicated significant differences in survival (p=0.043) and the mean life span (p=0.033) in male SAMP8 mice. Oligonol increased the mean life span and this was statistically significant. In the open-field locomotive test, the 7-week-old SAMP8 mice crossed more than 40 partitioned lines in 1min. At 48-week-old control untreated male SAMP8 crossed 2 lines. The Oligonol-treated 48-week-old male SAMP8 mice crossed 17 lines however. The improved locomotive activity was statistically significant even after 36weeks in the Oligonol-treated male SAMP8 but this was not the case throughout the time course of the study in the Oligonol-treated female SAMP8. Thus Oligonol treatment to SAMP8 mice modulated the severity of infection-dependent inflammation, prolonged life-span and significantly improved locomotive activity indicating potential benefit to aging-associated diseases such as Alzheimer's or Parkinson's diseases. This presents potential for further research to define infection-dependent inflammation associated with degenerative conditions and the molecular mechanism of dietary antioxidant protection.     

异基因骨髓移植小鼠联合输注IL-3/IL-6双基因转导的骨髓基质细胞促进造血恢复

探讨用逆转录病毒载体转入IL 3和IL 6双基因的骨髓基质细胞系QXMSC1IL 3/IL 6对异基因骨髓移植小鼠造血功能的促进作用。用逆转录病毒载体 (含小鼠IL 3cDNA ,人IL 6cDNA)分别转导到骨髓基质细胞系QXMSC1(H 2 d) ,构建骨髓基质细胞系QXMSC1IL 3/IL 6。供体小鼠BALB/c(H 2 d)骨髓去除骨髓中T细胞 ,受体小鼠C57BL/6(H 2 b)经γ射线致死量照射后 ,输入去除T细胞的供体骨髓 (1× 1 0 7/鼠 )的同时输入骨髓基质细胞QXMSC1IL 3/IL 6(5× 1 0 5/鼠 )。在骨髓移植后 2 0和 40天 ,分别检测骨髓移植小鼠外周血RBC ,WBC和骨髓有核细胞数及骨髓中CFU S ,CFU GM ,CFU E和CFU GEMM数。结果 :异基因骨髓移植共输入QXMSC1IL 3/IL 6基质细胞系可使异基因骨髓移植小鼠外周血RBC ,WBC数明显恢复 ,骨髓中有核细胞数 ,CFU S ,CFU GM ,CFU E和CFU GEMM明显增加。基质细胞系QXMSC1可作为有效的基因载体促进骨髓移植后造血功能重建。结论 :基质细胞转入细胞因子IL 3或IL 6基因可以进一步促进骨髓移植后造血功能重建 ,联合转导IL 3和IL 6有协同作用     

骨髓基质干细胞与骨缺损的修复

背景：骨髓基质干细胞来源广泛,分离方便,扩增迅速,具有多向分化潜能,适合自体移植的特点,同时是骨组织工程中合适的种子细胞。目的：总结分析采用骨髓基质干细胞作为种子细胞,并利用转基因或其复合支架修复骨缺损所具有的优势。方法：检索1990/2009西文生物医学期刊文献数据及CNKI数据库有关骨髓基质干细胞分离、培养,在骨缺损方面的应用,骨组织工程方面的文献,英文检索词为＂Bone marrow stem cells,Repairing,Bone defect＂,中文检索词为＂骨髓基质干细胞,修复,骨缺损＂。排除重复性研究,保留28篇进一步归纳总结。结果与结论：文章从骨髓基质干细胞作为种子细胞的优势,骨髓基质干细胞的分离和培养,骨髓基质干细胞的成骨诱导及骨髓基质干细胞修复骨缺损等方面进行了总结。利用骨髓基质干细胞作为种子细胞,与适当的支架材料结合,或用骨髓基质干细胞作为靶细胞,导入外源目的基因诱导成骨的基因治疗来修复骨缺损的方法,给骨缺损的治疗带来光明的前景。但同时也存在骨髓基质干细胞存化,骨髓基质干细胞的增殖、分化合适的条件,哪些因子能有效促进新骨形成,载体材料及体内植入方式,以及如何将骨组织工程与基因治疗的方法结合起来等问题需要进一步的研究。     

Development of large-scale manufacturing of adipose-derived stromal cells for clinical applications using bioreactors and human platelet lysate

In vitro expanded adipose-derived stromal cells (ASCs) are a useful resource for tissue regeneration. Translation of small-scale autologous cell production into a large-scale, allogeneic production process for clinical applications necessitates well-chosen raw materials and cell culture platform. We compare the use of clinical-grade human platelet lysate (hPL) and fetal bovine serum (FBS) as growth supplements for ASC expansion in the automated, closed hollow fibre quantum cell expansion system (bioreactor). Stromal vascular fractions were isolated from human subcutaneous abdominal fat. In average, 95?×?10⁶ cells were suspended in 10% FBS or 5% hPL medium, and loaded into a bioreactor coated with cryoprecipitate. ASCs (P0) were harvested, and 30?×?10⁶ ASCs were reloaded for continued expansion (P1). Feeding rate and time of harvest was guided by metabolic monitoring. Viability, sterility, purity, differentiation capacity, and genomic stability of ASCs P1 were determined. Cultivation of SVF in hPL medium for in average nine days, yielded 546?×?10⁶ ASCs compared to 111?×?10⁶ ASCs, after 17?days in FBS medium. ASCs P1 yields were in average 605?×?10⁶ ASCs (PD [population doublings]: 4.65) after six days in hPL medium, compared to 119?×?10⁶ ASCs (PD: 2.45) in FBS medium, after 21?days. ASCs fulfilled ISCT criteria and demonstrated genomic stability and sterility. The use of hPL as a growth supplement for ASCs expansion in the quantum cell expansion system provides an efficient expansion process compared to the use of FBS, while maintaining cell quality appropriate for clinical use. The described process is an obvious choice for manufacturing of large-scale allogeneic ASC products.