Muscle stem cells and model systems for their investigation.

Stem cells are characterized by their clonal ability both to generate differentiated progeny and to undergo self-renewal. Studies of adult mammalian organs have revealed stem cells in practically every tissue. In the adult skeletal muscle, satellite cells are the primary muscle stem cells, responsible for postnatal muscle growth, hypertrophy, and regeneration. In the past decade, several molecular markers have been found that identify satellite cells in quiescent and activated states. However, despite their prime importance, surprisingly little is known about the biology of satellite cells, as their analysis was for a long time hampered by a lack of genetically amenable experimental models where their properties can be dissected. Here, we review how the embryonic origin of satellite cells was discovered using chick and mouse model systems and discuss how cells from other sources can contribute to muscle regeneration. We present evidence for evolutionarily conserved properties of muscle stem cells and their identification in lower vertebrates and in the fruit fly. In Drosophila, muscle stem cells called adult muscle precursors (AMP) can be identified in embryos and in larvae by persistent expression of a myogenic basic helix-loop-helix factor Twist. AMP cells play a crucial role in the Drosophila life cycle, allowing de novo formation and regeneration of adult musculature during metamorphosis. Based on the premise that AMPs represent satellite-like cells of the fruit fly, important insight into the biology of vertebrate muscle stem cells can be gained from genetic analysis in Drosophila.     

When cells tell their neighbors which direction to divide.

Many cells must divide in specific orientations, yet for only a handful of cases do we have some understanding of how cells choose division orientations. We know of only a few cases where division orientations are controlled by specific cell-cell interactions. These cases are of interest, because they tell us something new and seemingly fundamental about how cells can function during development. Here, the evidence that interactions control division orientation in some cells of the early C. elegans embryo is presented, and what is known about how contact can regulate division orientation is discussed. Whether contact-mediated division orientation is a peculiarity of C. elegans or whether it may be more widespread is addressed.     

Hematopoietic stem cells and their precursors: developmental diversity and lineage relationships

Summary:  Within the context of the developing embryo, restrictions in cell lineage potential occur through cell–cell interactions and signaling molecules, leading to changes in genetic programs and to the emergence of disparate tissues containing functionally distinct cell types including somatic stem cells. Tissue maintenance in the adult is thought to occur through specific stem cells, and in the case of the hematopoietic system, through hematopoietic stem cells (HSCs). These cells arise in midgestation within the region of the embryo containing the dorsal aorta, gonads, and mesonephros (AGM) and are thought to maintain a distinct hematopoietic lineage-restricted fate. However, recent transplantation experiments suggest that within the adult, HSCs previously thought to be restricted can, under certain circumstances, display unexpected lineage potentials. With these surprising and controversial results, it is becoming apparent that a better understanding of the developmental processes, molecular programs and lineage relationships leading to the emergence of adult stem cells will provide insight into the incremental steps involved in lineage determination, and perhaps possibilities for the manipulated differentiation of stem cells. The most widely studied, accessible stem cell and cellular differentiation hierarchy is that of the hematopoietic system. With the issue of stem cell potential in the forefront, the focus of this review is on the development of the hematopoietic system: how HSCs arise in the embryo, the lineage relationships of hematopoietic cells as they are generated, and the identification of precursor cells fated to the hematopoietic lineage throughout ontogeny.     

The origin of intestinal stem cells in Drosophila

Renewing tissues in the adult organism such as the gastrointestinal (GI) epithelium depend on stem cells for epithelial maintenance and repair. Yet, little is known about the developmental origins of adult stem cells and their niches. Studies of Drosophila adult midgut precursors (AMPs), a population of endodermal progenitors, demonstrate that adult intestinal stem cells (ISCs) arise from the AMP lineage and provide insight into the stepwise process by which the adult midgut epithelium is established during development. Here, I review the current literature on AMPs, where local, inductive and long-range humoral signals have been found to control progenitor cell behavior. Future studies will be necessary to determine the precise mechanism by which adult intestinal stem cells are established in the endodermal lineage.     

Haemopoietic stem cells: their heterogeneity and regulation

The maintenance of the various blood cell populations in mammals is achieved by the proliferation and differentiation of precursor cells located primarily in the bone marrow. These precursor cells are all derived from a common haemopoietic stem cell population that is established during embryogenesis and functions for the lifetime of the organism. An overview is given of the various assay systems for haemopoietic stem cells, how these assays have contributed to understanding the considerable heterogeneity within the stem cell compartment, the regulation of stem cells and the development of the haemopoietic system.     

Neural stem cells: plasticity and their transdifferentiation potential

The presence of resident stem cells in adult tissues is of fundamental importance for the maintenance of their structural and functional integrity. In fact, throughout life, somatic stem cells attend to the critical function of substituting terminally differentiated cells lost to physiological turnover, injury or disease. Thence, one of the basic dogmata in tissue biology holds that the differentiation potential of an adult stem cell is restricted to the generation of the mature cell lineages found in the tissue to which the stem cell belongs. A plethora of recent evidences from many groups, including ours, is now providing evidence that adult stem cells may possess a broader differentiation repertoire than expected and that their fate potential may not be as tissue specific as once thought. The initial example of an unforeseen, trans-germ layer plasticity - that seems now to emerge as a prototypic functional trait of various somatic stem cells of different origin - has come from the reported awakening of a latent hemopoietic developmental capacity in stem cells isolated from the adult mammalian brain following their transplantation into sub-lethally irradiated mice. More recently, it has been shown that adult neural stem cells can differentiate into a wide array of bodily cells of different origin when injected into the blastocyst and into myogenic cells when transplanted into the adult regenerating skeletal muscle. Moreover, bone marrow stem cells can now give rise to skeletal muscle, hepatic and brain cells, whereas muscle precursors can generate blood cells. In this article, we review some of the basic notions regarding the functional properties of the adult neural stem cells and discuss findings in the expanding area of trans-germ layer conversion, with emphasis on the neural stem cell.     

滤泡辅助性T细胞的主要特性及其活化的信号转导途径

滤泡辅助性T细胞能够迁移到淋巴滤泡,参与促进生发中心的形成、维持,浆细胞(长寿的和短寿的)和记忆B细胞的产生和抗体类别转换,在辅助B淋巴细胞产生抗体中起着重要作用,是近年来发现的一种新CD4+辅助性T细胞亚群。本文将重点围绕目前综述涉及不多的信号通路调控,综述滤泡辅助性T细胞的发育分化、主要特性及与疾病的关系。     

三维微小凹图式的制备及其与C17.2神经干细胞的复合

以紫外光光刻及氢氟酸湿法蚀刻加工硅阳模,采用基于聚二甲基硅氧烷(PDMS)的软光刻技术制备9种不同结构尺寸的聚乳酸-羟基乙酸共聚物(PLGA)和PMDS三维微小凹图式。PLGA及PDMS三维微小凹图式经等离子氧蚀刻和多聚赖氨酸裱衬处理后进行C17.2神经干细胞培养。随着在图式上培养时细胞的增殖,C17.2神经干细胞逐渐在微小凹中聚集,表现出明显的三维生长行为;通过羧基荧光素乙酰乙酸琥珀酰亚胺酯(CFDA-SE)染色后进行激光共聚焦显微扫描与三维重构,显示大部分细胞生长于微小凹中离底面30～90μm的区间内;免疫荧光结果显示C17.2神经干细胞在三维微结构中复合培养2d后呈现均一的巢蛋白(Nestin)阳性。结论:本文设计的微小凹图式适用于C17.2神经干细胞的三维培养及后续的分化研究,细胞于微小凹图式培养过程中可以保持均一的干细胞特性。     

无血清培养人胚胎干细胞的体系研究与进展

背景：人胚胎干细胞有永久自我更新和向外、中和内胚层分化的能力,具有重要的基础研究价值和临床应用前景,可应用于再生医学、药物毒性筛选、早期胚胎发育、细胞移植治疗和基因治疗等领域,但是在限定条件下的大规模细胞培养、控制和定向分化技术的发展仍然具有实质性的挑战。在治疗应用方面,许多研究者试图建立可在无异源限定性和规模化培养体系中维持细胞多能性的培养方法。 目的：综述近几年报道的人胚胎干细胞的无血清培养体系研究进展,重点是在发展适于治疗用途的无血清和无异源培养体系进程中,面临的挑战和进步。 方法：应用计算机检索CNKI和PubMed学术数据库中2008至2015年关于人胚胎干细胞无血清培养的文章,排除观点重复和陈旧的文章,最后对58篇文章进行归纳汇总。 结果与结论：研究者已经在使用人源性饲养层细胞、无饲养层基质及限定性成分培养基上做了许多尝试,旨在选择合适的基质和培养基,使异源性成分达到最小化或者不使用,以求获得临床级别的细胞株。但是目前的细胞制品距离临床应用还有很大的距离,仍有很多问题需要解决,比如细胞制品的规范化、标准化、个性化定制等。相信随着干细胞研究和行业的规范,胚胎干细胞产品将会在临床得到广泛应用。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

Evolution and genomic signatures of spontaneous somatic mutation in Drosophila intestinal stem cells

Spontaneous mutations can alter tissue dynamics and lead to cancer initiation. Although large-scale sequencing projects have illuminated processes that influence somatic mutation and subsequent tumor evolution, the mutational dynamics operating in the very early stages of cancer development are currently not well understood. To explore mutational processes in the early stages of cancer evolution, we exploited neoplasia arising spontaneously in the Drosophila intestine. Analysing whole-genome sequencing data with a dedicated bioinformatic pipeline, we found neoplasia formation to be driven largely through the inactivation of Notch by structural variants, many of which involve highly complex genomic rearrangements. The genome-wide mutational burden in neoplasia was found to be similar to that of several human cancers. Finally, we identified genomic features associated with spontaneous mutation, and defined the evolutionary dynamics and mutational landscape operating within intestinal neoplasia over the short lifespan of the adult fly. Our findings provide unique insight into mutational dynamics operating over a short timescale in the genetic model system, Drosophila melanogaster.

The accumulation of mutations in somatic tissues plays a major role in cancer and is proposed to contribute to aging (Al Zouabi and Bardin 2020). Although the majority of mutations acquired throughout life are harmless, some alter cellular fitness and become subject to the selective forces operative in cells and tissues. Mutations that confer a selective advantage can lead to the formation of a clonal population of mutant cells under positive selection. Such events, termed driver mutations, underscore cancer formation and, as such, have been the subject of extensive investigation (Bailey et al. 2018; Alexandrov et al. 2020; Rheinbay et al. 2020; The ICGC/TCGA Pan-Cancer Analysis of Whole Genomes Consortium 2020). These initiating mutations are thought to arise in normal cells and can therefore provide key insights into the mutational processes at play in precancerous states. Large-scale sequencing projects have detailed the mutational burdens of human cancer genomes and have revealed the repertoire of somatic mutations driving cancer formation, illuminating the biological processes underlying somatic mutation. Cancer genomes, however, represent the end-point of a long evolutionary process that shapes the mutational landscape of tumors. Similarly, the mutations recently described to arise in aged normal cells and early-stage cancers represent the result of many years of selective pressure and mutational dynamics (Martincorena et al. 2015, 2018; Lee-Six et al. 2019; Moore et al. 2020; Yokoyama et al. 2019). Knowledge of mutational processes operative in the very earliest stages of cancer is therefore currently incomplete.Our previous work has established the Drosophila midgut as an excellent model system for understanding somatic mutation in an adult tissue-specific stem cell population (Siudeja et al. 2015). In this tissue, intestinal stem cells (ISCs) self-renew and divide to give rise to two differentiated cell types: absorptive enterocytes (ECs) and secretory enteroendocrine cells (EEs) (Micchelli and Perrimon 2006; Ohlstein and Spradling 2006). We have previously shown that during aging, 12% of wild-type male flies harbor spontaneous mutations that inactivate the X-linked tumor-suppressor gene Notch, driving hyperproliferation of ISCs and EEs and resulting in neoplasm formation (Siudeja et al. 2015).Here, we take advantage of the spontaneous formation of neoplasia in the intestine of the fruit fly to investigate the processes underlying early somatic mutation and evolution within a clonal cell population.     

Nonrandom sister chromatid segregation of sex chromosomes in Drosophila male germline stem cells

Sister chromatids are the product of DNA replication, which is assumed to be a very precise process. Therefore, sister chromatids should be exact copies of each other. However, reports have indicated that sister chromatids are segregated nonrandomly during cell division, suggesting that sister chromatids are not the same, although their DNA sequences are the same. Researchers have speculated that stem cells may retain template strands to avoid replication-induced mutations. An alternative proposal is that cells may segregate distinct epigenetic information carried on sister chromatids. Recently, we found that Drosophila male germline stem cells segregate sister chromatids of X and Y chromosomes with a strong bias. We discuss this finding in relation to existing models for nonrandom sister chromatid segregation.     

From pathogens to microbiota: How Drosophila intestinal stem cells react to gut microbes

The intestine acts as one of the interfaces between an organism and its external environment. As the primary digestive organ, it is constantly exposed to a multitude of stresses as it processes and absorbs nutrients. Among these is the recurring damage induced by ingested pathogenic and commensal microorganisms. Both the bacterial activity and immune response itself can result in the loss of epithelial cells, which subsequently requires replacement. In the Drosophila midgut, this regenerative role is fulfilled by intestinal stem cells (ISCs). Microbes not only trigger cell loss and replacement, but also modify intestinal and whole organism physiology, thus modulating ISC activity. Regulation of ISCs is integrated through a complex network of signaling pathways initiated by other gut cell populations, including enterocytes, enteroblasts, enteroendocrine and visceral muscles cells. The gut also receives signals from circulating immune cells, the hemocytes, to properly respond against infection. This review summarizes the types of gut microbes found in Drosophila, mechanisms for their elimination, and provides an integrated view of the signaling pathways that regulate tissue renewal in the midgut.     

Clonogenic analysis reveals reserve stem cells in postnatal mammals: I. Pluripotent mesenchymal stem cells

Clonal populations of lineage-uncommitted pluripotent mesenchymal stem cells have been identified in prenatal avians and rodents. These cells reside in the connective tissue matrices of many organs and tissues. They demonstrate extended capabilities for self-renewal and the ability to differentiate into multiple separate tissues within the mesodermal germ line. This study was designed to determine whether such cells are present in the connective tissues of postnatal mammals. This report describes a cell clone derived by isolation from postnatal rat connective tissues, cryopreservation, extended propagation, and serial dilution clonogenic analysis. In the undifferentiated state, this clone demonstrates a high nuclear-to-cytoplasmic ratio and extended capacity for self-renewal. Subsequent morphological, histochemical, and immunochemical analysis after the induction of differentiation revealed phenotypic markers characteristic of multiple cell types of mesodermal origin, such as skeletal muscle, smooth muscle, fat cells, cartilage, and bone. These results indicate that this clone consists of pluripotent mesenchymal stem cells. This report demonstrates that clonal populations of reserve stem cells are present in mammals after birth. Potential roles for such cells in the maintenance, repair, and regeneration of mesodermal tissues are discussed.     

Breast cancer,stem cells and sex hormones. Part 2: The impact of the reproductive years and pregnancy

The primitive breast develops in utero and during infancy breast growth largely parallels the growth of the child. At puberty, the GnRH pulse generator starts up, initially with just 1–2 pulses daily. This results in very small amounts of unopposed estrogen being secreted by the ovary. As the GnRH pulse generator matures, ovarian secretion of estrogen increases. The pubertal breast responds to this increasing estrogen drive. Breast glandular increase in size is mostly due to growth and division of the primary ducts. Eventually, the terminal buds give rise alveolar buds which tend to cluster around a terminal duct. Lobule formation begins in the first 2 years that follow menarche. With the onset of ovulation, breast mitotic activity increases and is usually maximal in the luteal phase. The final stage of breast maturation occurs during the first full-term pregnancy. The breast undergoes marked changes in preparation for breast feeding. There is evidence that breast SC number decreases during that first pregnancy. Also, the remaining SC undergo significant change which seems to render them less likely to undergo malignant change. These alterations to breast SC number and function may explain, at least in part, why early first pregnancy reduces the risk of breast cancer later in life.