Acknowledgments

This study was supported by a Grant-in-aid ( 15390636) for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan. This work was also carried out in courtesy of the Research Center for Molecular Medicine, Graduate School of Biomedical 1 Pittenger, MF Mackay, AM Beck, SC Jaiswal, RK Douglas, R Mosca, JD Moorman, MA Simonetti, DW Craig, S Marshak, DR. Multilineage potential of adult human mesenchymal stem cells. Science, 1999, 284, 143-147. 2 Bell, E Ivarsson, B...

References

Biology and management of acquired severe aplastic anaemia. Current opinion in Oncology, 1998, 10, 95-99. 2 Stohlman, F. Jr. Aplastic Anaemia. Blood, 1972, 4, 282. 3 Ehrlich, P. Uber einen Fall von Anamie mit Bemerkungen uber regenerative Veranderungen des Knochenmarks. Charite-Annalen. 1888, 13, 300. 4 Chauffard, M. Un cas d'anaemie pernicieuse aplastique. Bull. Mem. Soc. Med. Hop. Paris, 1904, 21, 313. 5 Juneja, HS Lee, S Gardner, FH. Human long term bone marrow cultures in...

Summary and Future Perspectives

Various detection methods have been described to study the functional relevance of renal infiltrating BMDC. Provided that the essential prerequisites of enzymatic staining are met, all above mentioned reporters would seem applicable and useful for tracing BMDC in renal disease models. Some have, however, advantages over others. EGFP and luciferase both can be detected in living cells, which is an advantage for functional studies. The advantage of EGFP as a reporter molecule over luciferase is...

Stem Stromal Imbalance

A collosum amount of literature have been accumulated concerning the role of marrow microenvironment for maintaining the proper sternness within the bone marrow 27 . Current studies in our laboratory also furnished that the microenvironmental factors can stimulate the stem cell proliferation and function 28 . These observations simultaneously support the view that in Aplastic Anaemia the microenvironmental support is also disorganized 29 . In experimentally induced Aplastic Anaemia, such...

Statistical Analysis

Experiments were performed at least in triplicate and expressed as mean standard deviations. The student's t test was used for statistical analysis. Table 1. Primer Sequences Used in Chondrogenic Differentiation Table 1. Primer Sequences Used in Chondrogenic Differentiation Table 2. Primer and probe Sequences Used in Osteogenic Differentiation Hs00173720_m1 Applied Biosystem s Hs00740632_gH Applied Biosystems Hs00164004_m1 Applied Biosystem s S'-TGGACGAGGCAAGAGTTTCA-S'...

Transcriptional Factors and Networks Controlling Self Renewal of hES Cells

Several key transcription factors have been implicated to play a pivotal role in maintenance of hESCs in an undifferentiated state. Among them, Oct3 4 and Nanog are perhaps the two most well-known homeobox transcription factors that are not only expressed both in mouse and human ESC, but are essential in maintaining pluripotency Chambers and Smith, 2004 Zaehres et al., 2005 . However, there is limited knowledge about the upstream regulators that mediate the transcription of these genes. Oct3 4...

MicroRNA and Stem Cells Differentiation

Cell Differentiation Factors

MiRNAs regulate cancer cell differentiation 30, 35 , control over lineage determination in hematopoiesis 29 , and neuronal stem cell differentiation 36 . miR-181 was preferentially expressed in the B-lymphoid cells of mouse bone marrow. Its ectopic expression in hematopoietic stem progenitor cells led to an increased differentiation of B-lineage cells. Kanellopoulou et al. 34 recently observed that DCRA A ES cells express similar level of Oct-4 compared with DCR wildtype ES cells. These DCRA A...

Ex Vivo Labeling of bmdc

Besides the use of bone marrow transplantation to study the role of BMDC in renal disease models, local or systemic infusion of BMDC, or a sub-set of BMDC, has been used. This approach does not require total body irradiation and is more relevant in pre-clinical, rather than experimental, studies. However, since the native bone marrow is not replaced by bone marrow transplantation, the infused BMDC will compete with physiologically recruited BMDC, which might hamper the interpretation of BMDC...

Conclusion

The intriguing fact that resounds again and again is that the condition of stem stromal balance is essential in the formation and the maintenance of a healthy niche which inter alia serves as the complete machinery for haematopoietic generation, wherein different cytokines and growth factors serve as the fuel of the machinery. Thus we emphasize that supplementation of only fuel i.e. cytokines and growth factors may not activate the intricate damages in the machinery as we find in experimental...

Stem Cell Research Mexico Df

Natural Killer Cell Receptor NKG2A HLA-E Interaction Dependent Differential Thymopoiesis of Hematopoietic Progenitor Cells Influences the Outcome of HIV Edmond J. Yunis a, Viviana Romeroa, Felipe Diaz-Gifferoa, Joaquin Zu igaa'b and Prasad Kokab aDepartment of Cancer Immunology and AIDS, Dana-Farber Cancer Institute and Department of Pathology, Harvard Medical School, Boston MA b Laboratory of Immunobiology and Genetics, Instituto Nacional de Enfermedades Respiratorias, Mexico D.F. , Mexico. c...

Cytokines Involved in Stemness Regulation in HSCs

A number of cytokines regulate growth, differentiation, and survival of HSCs both positively and negatively. Among these, stem cell factor SCF , Flt3 ligand FL , thrombopoietin TPO , interleukin-3 IL-3 , and IL-6 are known to promote the growth of HSCs in vitro 18-20 . In fact, Sl Sl and W W mice each having homozygous defect in the SCF gene and its receptor c-kit gene reveal severe anemia 21 . Also, total number of HSCs was reduced in the BM of c-mpl TPO receptor -null mice 22 . In addition,...

Aplastic Anaemia

Stem Cell Research and Application Unit Department of Biochemistry and Medical Biotechnology, School of Tropical Medicine Aplastic Anaemia is correlated with Stem Cell disorder, the detail mechanism of which still awaits extensive evaluation. Apart from Inherited Aplastic Anaemia, the Acquired Aplastic Anaemia, has also been considered to involve stem cell deficiency or disorder or more currently considered disorganized stem cell niche. The manifestations in such types of Aplastic Anaemia are...

Immune Functions in hiv Infection

Cognate immunity has been extensively studied involving primarily CD4 and CD8 T cells interacting with MHC alleles presenting HIV-1 peptides on antigen presenting cells 612 . Such cells may mediate protection by production of cytotoxic cells or cytokines such as IFN gamma and IL-12 13 . Macrophages 14 , DC and plasmocytoid DCs are type 1 IFN secretion inducers of IL-12. However, several viruses escape MHC class I restricted cytotoxic T lymphocytes CTL responses by down-regulating their...

Transplantation

The most commonly used technique to track BMDC and study their role of BMDC after renal injury is transplantation of bone marrow from rodents that are genetically distinguishable from the recipient, into bone marrow-ablated recipients. After bone marrow transplantation, the transplanted bone marrow cells stably reconstitute the recipient's bone marrow compartment. In practice, the efficiency of bone marrow ablation and reconstitution with donor bone marrow is never 100 and a certain degree of...

Preface

The two broad categories of mammalian stem cells exist embryonic stem cells, derived from blastocysts, and adult stem cells, which are found in adult tissues. In a developing embryo, stem cells are able to differentiate into all of the specialised embryonic tissues. In adult organisms, stem cells and progenitor cells act as a repair system for the body, replenishing specialised cells. As stem cells can be readily grown and transformed into specialised tissues such as muscles or nerves through...

Discussion

In the present report, a purified autologous MSC population was transplanted to an AMI patient. The cell preparation and administration protocol were different from recently reported clinical trials 7-9 . Bone marrow consists of a nonhomogeneous population of cells and contains a stem cell subpopulation. The stem cell fraction of bone marrow is mainly composed of hematopoietic stem cell HSC . MSCs are non-HSC fraction and represent only 0.01-0.0001 of the nucleated cells in adult human bone...

HESCs Respond to the hBMSC Transwell CoCulture System and Express Osteogenic Specific Markers

Osteoblastic lineage commitment can be observed by the expression of bone specific transcription factors runx2 and osterix, and with collagen type I production. Runx2 is a homolog of the Drosophila Runt protein that acts as a transcriptional regulator of osteoblast differentiation, and osterix is a zinc finger containing transcription factor required for osteoblastic differentiation that acts downstream of runx2 17, 18 . Collagen type I is the most abundant protein found in bone ECM 19 ....

HESChBMSC Transwell CoCulture

The hBMSCs were isolated from patients at the University of Michigan using IRB approved protocols. The hBMSCs were plated at 2,000 cm2 onto transwell inserts 0.4 m pore, Corning, Corning, NY and allowed to differentiate for 7-24 days in OS with complete medium changes every 48 hours. Two days prior to adding the hBMSC transwell inserts, the hESCs were either made into EBs or manually passaged directly from MEFs omitting EB formation and then seeded onto gelatin-coated 6-well plates. After 7, 14...

Materials and Methods

Human blood samples and fetal tissue collections for research purpose were approved by the Research Ethics Committees of Institute of Zoology, Chinese Academy of Sciences. National guidelines were complied with the regulation in relation to the use of fetal tissues only for research. Antibodies against human antigens including CD29, CD34, CD44, CD45, CD31, CD58, HLA-DR, and the isotype antibodies were purchased from Becton Dickinson except for CD105, which is purchased from RandD systems....

Introduction

Hematopoietic stem cells HSCs are rare, pluripotent cells that give rise to all hematopoietic cell types and possess extensive proliferative and self-renewal capabilities 1, 2 . The regulation of self-renewal and differentiation of HSCs requires a specific microenvironment of surrounding cells known as the stem cell niche 3, 4, 5, 6, 7 , which contains hematopoietic-supporting stromal cells and extracellular matrix 8, 9 . During fetal development when hematopoiesis is transiently localized to...

Semi Quantitative Real Time Reverse Transcription Polymerase Chain Reaction Real Time Rtpcr Analysis

Total RNA was isolated from the gel cultures using a guanidine thiocyanate method 21 . A single strand cDNA was synthesized from 1 g of total RNA using Oligo dT 20 primer Toyobo, Osaka, Japan and a Rever Tra Ace-a first strand cDNA synthesis kit Toyobo . The mRNA levels were determined by semi-quantitative real time RT-PCR analysis, using a SYBR Green PCR master mix Applied Biosystems, Foster City, CA or a TaqMan Universal PCR master mix Applied Biosystems with an automated fluorometer ABI...

Contents

Human Embryonic Stem Cells Undergo Osteogenic Differentiation in Human Bone Marrow Stromal Cell Microenvironments Wilbur Tong, Shelley E. Brown and Paul H. Krebsbach Hyaluronic Acid Promotes Osteochondrogenic Differentiation from Mesenchymal Stem Cells in a Three-Dimensional Culture System Kiyoshi Yoneno, Eiji Tanaka, Shigeru Ohno, Kotaro Tanimoto, Kobun Honda, Nobuaki Tanaka, Yu-Yu Lin, Yuki Tanne, Satoru Okuma and Kazuo Tanne Cell Cycle Regulation in Maintaining Immortality of Embryonic Stem...

Mineralization Assays by Alizarin Red S and von Kossa Staining

Cell cultures were fixed in 4 paraformaldehyde for 30 minutes, washed twice with PBS and then stained. Alizarin Red S Sigma staining was used to determine the presence of mineralized nodules. Fixed cells were incubated for 1 hour in 1 Alizarin Red S solution and then washed twice with water. Von Kossa staining was used to determine the presence of phosphate. Fixed cells were incubated for 1 hour in 5 AgNO3 solution and exposed to bright light for at least 30 minutes. For mineralization...