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Cloning Stem Cells. 2002;4(1):9-19.
Production of transgenic chimera rabbit fetuses using somatic cell nuclear transfer.

Matsuda J, Takahashi S, Ohkoshi K, Kaminaka K, Kaminaka S, Nozaki C, Maeda H, Tokunaga T.

The Chemo-Sero-Therapeutic Research Institute, Kikuchi Research Center, Kumamoto, Japan. matsuda-jaketsuken.or.jp

We produced aggregate chimeric embryos between blastomeres from the somatic cell nuclear transfer (SCNT) embryos and blastomeres from normal embryos. The SCNT embryos were produced by fusing enucleated oocytes with GFP gene introduced fibroblast cells, which were derived from a day 16 fetus. GFP gene-introduced fibroblast cells were cultured and passaged four to 12 times over a period of 45-79 days before SCNT. After transferring them into pseudopregnant recipient rabbits, the 15-day postcoitus fetuses were collected. We examined the existence of the cells derived from SCNT embryos in the fetus stage of pregnancy to detect the GFP gene. Fetuses that were not collected continued to develop into newborn rabbits. Two hundred and thirty-six chimeric embryos were produced using 39 SCNT morula stage embryos, and these embryos were transferred to 11 recipient rabbits. As a result, 27 normally developed and 16 degenerated concepti were obtained. The GFP gene-positive signals were detected in one of the fetuses, two of the placentae, and two of the degenerated concepti. In this study, we found that the rabbit SCNT embryos have the ability to develop and differentiate in vivo. We also demonstrated a novel method of producing a transgenic rabbit using SCNT.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12006152&dopt=Abstract

FEBS Lett. 2002 Nov 20;531(3):389-96.
Isolation of embryonic stem-like cells from equine blastocysts and their differentiation in vitro.

Saito S, Ugai H, Sawai K, Yamamoto Y, Minamihashi A, Kurosaka K, Kobayashi Y, Murata T, Obata Y, Yokoyama K.

Gene Engineering Division, BioResource Center, RIKEN, Tsukuba, 305-0074, Ibaraki, Japan.

Embryonic stem (ES) cells are pluripotent cells with the potential capacity to generate any type of cell. We describe here the isolation of pluripotent ES-like cells from equine blastocysts that have been frozen and thawed. Our two lines of ES-like cells (E-1 and E-2) appear to maintain a normal diploid karyotype indefinitely in culture in vitro and to express markers that are characteristic of ES cells from mice, namely, alkaline phosphatase, stage-specific embryonic antigen-1, STAT-3 and Oct 4. After culture of equine ES-like cells in vitro for more than 17 passages, some ES-like cells differentiated to neural precursor cells in the presence of basic fibroblast growth factor (bFGF), epidermal growth factor and platelet-derived growth factor. We also developed a protocol that resulted in the differentiation of ES-like cells in vitro to hematopoietic and endothelial cell lineages in response to bFGF, stem cell factor and oncostatin M. Our observations set the stage for future developments that may allow the use of equine ES-like cells for the treatment of neurological and hematopoietic disorders.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12435581&dopt=Abstract

Cytotherapy. 2002;4(2):109-18.
Ex vivo expansion of human UC blood primitive hematopoietic progenitors and transplantable stem cells using human primary BM stromal cells and human AB serum.

Yamaguchi M, Hirayama F, Murahashi H, Azuma H, Sato N, Miyazaki H, Fukazawa K, Sawada K, Koike T, Kuwabara M, Ikeda H, Ikebuchi K.

Hokkaido Red Cross Blood Center, Sapporo, Japan.

BACKGROUND: In vitro maintenance and expansion of human hematopoietic stem cells is crucial for many clinical applications, and investigators have been using xenogeneic, especially murine, stromal cells for stem-cell expansion. In addition, many such culture systems utilize FCS-containing medium or serum-free medium that contains human- or animal-derived proteins. However, the possible transmission of infectious diseases has led to a debate about the safety of the delivery of grafts expanded in culture using cells and proteins of allogeneic or xenogeneic origin. Using primary human BM stromal cells, we have established an AB serum-based co-culture system to expand human primitive progenitors and transplantable stem cells. METHODS: Cord blood CD34+ cells were cultured on a monolayer of human BM-derived primary stromal cells with thrombopoietin (TPO), stem-cell factor (SCF) and flt3/flk2 ligand (FL) in the presence of either FCS or AB serum. One to three weeks later, cells were examined for total cells, CD34+ cells, CD34+ CD38- cells, and clonogenic progenitors. SCID mouse reconstituting cell (SRC) activity was also studied. RESULTS: Three weeks of culture with TPO, SCF, and FL supported more than a 250-fold expansion of CD34+ cells, CD34+ CD38- cells and CFU-C, regardless of the kind of serum used. SRC assay revealed that transplantable stem cells were moderately expanded as well. DISCUSSION: This ex vivo expansion system should prove valuable in clinical settings in which stromal cells and serum are available from recipients or stem-cell donors.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12006206&dopt=Abstract

Cytotherapy. 2002;4(2):127-35.
The storage and re-infusion of autologous blood and BM as back-up following failed primary hematopoietic stem-cell transplantation: a survey of European practice.

Pottinger B, Walker M, Campbell M, Holyoake TL, Franklin IM, Cook G.

Bone Marrow Transplant Unit, Royal Infirmary, Glasgow, UK.

BACKGROUND: Traditionally, autologous BM or PBSC have been stored as a secondary source ('back-up') of hematopoietic stem cells (HSC) prior to allogeneic and autologous HSC transplantation. METHOD: We conducted an audit of a single transplant center practice for providing back-up HSC and compared this practice with other European centers. Laboratory records relating to the collection and re-infusion of consecutive HSC harvests were reviewed for 515 transplants (300 autologous and 215 allogeneic HSC transplants). RESULTS: In our experience, 2.3% (five of 215) of allogeneic HSC transplants required secondary HSC rescue for failure to engraft or graft failure (MUD, n = 2; un-manipulated sibling BMT, n = 1; T-cell depleted sibling BMT, n = 2). For autologous transplants, 4.7% (14 of 300) required rescue due to failure to engraft or late graft failure (ABMT for AML, n = 8; CD34+ cell selection/ex vivo expanded, n = 4; ABMT/PBSCT, n = 2). Among the European centers, 69.7% replied to a postal questionnaire, demonstrating that 81.4% and 45.6% of centers stored a secondary HSC source for manipulated and unmanipulated MUD BMT, respectively; 50% and 11.6% of centers stored a secondary source of HSC for manipulated and unmanipulated matched sibling BMT, respectively; 36.4% and 12.7% of centers stored HSC for manipulated and unmanipulated matched sibling PBSCT, respectively. In the autologous setting, 15.2% and 62.1% of centers stored back-up for unmanipulated and manipulated BMT, respectively and 19.5% and 68.5% stored back-up for unmanipulated and manipulated PBSCT, respectively, when myeloablative conditioning regimens were used. DISCUSSION: These data suggest that a small minority of patients require a secondary source of HSC rescue, most commonly in transplants with higher risk of graft failure. This is reflected in the practice across Europe of storing 'back-up' HSC. Guidelines should accommodate the need for storage of a secondary source of HSC only in those transplants associated with a higher risk of graft failure, especially in relation to graft engineering.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12006208&dopt=Abstract

J Biol Chem. 2002 Aug 2;277(31):27629-35. Epub 2002 May 10.
Variant forms of alpha-fetoprotein transcripts expressed in human hematopoietic progenitors. Implications for their developmental potential towards endoderm.

Kubota H, Storms RW, Reid LM.

Department of Cell and Molecular Physiology and Program in Molecular Biology and Biotechnology, University of North Carolina at Chapel Hill, School of Medicine, Chapel Hill, North Carolina 27599-7038, USA. kubotet.upenn.edu

Hematopoietic stem cells have been identified as multipotent cells that give rise to all adult hematopoietic lineages. Although the hematopoietic lineage is derived from the mesodermal germ layer in the embryo, recent data suggest that bone marrow cells with an antigenic profile consistent with that of hematopoietic stem cells can also differentiate to cell types of the endodermal lineages, such as hepatocytes. However, the molecular mechanisms associated with these events are entirely unknown. For decades, alpha-fetoprotein (AFP) has been used as a differentiation marker for endodermal cells, because it was thought that the transcription of AFP mRNA is tightly regulated in a developmental and tissue-specific process. In this report we describe two new variant forms of AFP transcripts in human hematopoietic progenitors that are not expressed in mature cells. The variant AFP (vAFP) cDNA sequences isolated from a multipotent hematopoietic cell line, K562, revealed that the vAFP differed from the authentic transcript, consisting of 15 exons, by replacing exon 1 of AFP with one or two exons located in the 5'-untranslated region of the AFP gene. In addition to the K562 cell line, vAFP transcripts were detected in normal bone marrow, thymus, and brain but were not detected in normal spleen, intestine, liver, or the hepatocellular carcinoma cell line, HepG2. This suggests expression in normal hematopoietic progenitors. This hypothesis was confirmed by the finding that CD34(+)Lin(-) hematopoietic progenitor cells purified from cord blood by flow cytometric sorting also expressed the variant transcripts. These results suggest that some hematopoietic progenitors are in a state that permits them to express certain types of transcripts that have been considered unique to endoderm.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12006569&dopt=Abstract

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Hair Loss, or alopecia is a concern for increasing number of folks in aging society. Loss of hair is a visible problem, and affects the appearance and changes identity of a person.
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Hair growth is a sophisticated biological process, which has not yet been completely understood. A multitude of therapeutic measures, including drugs, surgery, and suppelements have been made available, and used. However, due to the heterogeneity in the underlying cause, there is no perfect cure for all hair loss cases. Most of chemical drugs and hair transplantation surgeries are not free from varying degrees of undesirable side effects on health.

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