Regeneration of Hematopoietic Microenvironment (Bone Marrow) Using Adipose-Derived Stem Cells - The 2nd Report.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4304-4304
Author(s):  
Rei Ogawa ◽  
Juri Fujimura ◽  
Hiroshi Mizuno ◽  
Hiko Hyakusoku ◽  
Takashi Shimada
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
Transgenic Mice ◽  
Bone Matrix ◽  
Hematopoietic Cells ◽  
High Porosity ◽  
Adipose Derived Stem Cells ◽  
Bone Marrow Diseases ◽  
Bone Marrow Regeneration

Abstract Backgrounds: We have reported bone marrow regeneration using adipose-derived stem cells (ASCs) since 2003. In our study using mice, we showed that not only bone matrix but also bone marrow could be regenerated subcutaneously in vivo. Transplanted ASCs proliferated and differentiated into osteoblasts in micropores on the surface of scaffolds made of hydroxyapatites (HA). We have not yet obtained incontrovertible proof that the ASCs differentiated into hematopoietic cells. However, we could regenerate a subcutaneous microenvironment in which hematopoietic cells survived, which may lead to new treatments for fibrotic bone marrow diseases, such as idiopathic myelofibrosis and osteopetrosis. Purposes: In this report, we examined the optimal shapes of scaffolds for bone and bone marrow regeneration using ASCs. Methods: Taking advantage of homogeneously marked cells from green fluorescent protein (GFP) transgenic mice, ASCs were isolated from the inguinal fat pads of GFP transgenic mice. ASCs were cultured in vitro and after three passages were seeded into the small pores of scaffolds of various shapes made of HA. The ASC-containing scaffolds were then subsequently implanted into immunocompetent mice subcutaneously. Two months later, the scaffolds were extirpated for histological, immunohistochemical and flow cytometry analyses. Results: Histological examination showed that the pores were filled with bone matrix and that the bone marrow was composed of adipocytes, hematopoietic cells, and vasculatures. Bone marrow regeneration on small, thin scaffolds of high porosity was better than on other scaffolds. Immunohistochemical analysis confirmed that the GFP+ ASCs that had differentiated into osteoblasts were composed of bone matrix. Flow cytometry analyses showed that the bone marrow was composed of blood cells, including populations of monocytes, lymphocytes and granulocytes. Conclusions: Bone marrow engineering has great potential for hematopoietic disease therapy. The regeneration of the hematopoietic system using ASCs might be useful in the future for treating hematopoietic stem cell diseases such as leukemia as well as fibrotic bone marrow diseases. We plan to continue looking for suitable scaffolds and methods for bone and bone marrow regeneration using ASCs.

Hematopoietic Microenvironment Regeneration Using Adipose-Derived Stem Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4243-4243
Author(s):  
Rei Ogawa ◽  
Juri Fujimura ◽  
Hiroshi Mizuno ◽  
Hiko Hyakusoku ◽  
Takashi Shimada
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
Transgenic Mice ◽  
Bone Matrix ◽  
Hematopoietic Cells ◽  
High Porosity ◽  
Adipose Derived Stem Cells ◽  
Hematopoietic Microenvironment ◽  
Bone Marrow Diseases

Abstract Backgrounds: We have studied hematopoietic microenvironment regeneration using adipose-derived stem cells (ASCs) harvested from mice. We showed that not only bone matrix but also bone marrow could be regenerated subcutaneously, in vivo. Transplanted ASCs proliferated and differentiated into osteoblasts in micropores on the surface of scaffolds made of hydroxyapatites (HA). We could regenerate a subcutaneous microenvironment in which hematopoietic cells survived, which may lead to new treatments for fibrotic bone marrow diseases, such as idiopathic myelofibrosis and osteopetrosis. Objective: In this report, we examined the optimal shapes of scaffolds for bone and hematopoietic microenvironment regeneration using ASCs. Methods: Taking advantage of homogeneously marked cells from green fluorescent protein (GFP) transgenic mice, ASCs were isolated from the inguinal fat pads of GFP transgenic mice. ASCs were cultured in vitro and after three passages were seeded into the small pores of scaffolds of various shapes made of HA. The ASC-containing scaffolds were subsequently implanted into isogenic mice subcutaneously. Two months later, the scaffolds were extirpated for histological, immunohistochemical and flow cytometry analyses. Results: Histological examination showed that the pores were filled with bone matrix and that the bone marrow was composed of adipocytes, hematopoietic cells, and vasculatures. Bone marrow regeneration on small, thin scaffolds of high porosity was better than on other scaffolds. Immunohistochemical analysis confirmed that the GFP+ ASCs that had differentiated into osteoblasts were composed of bone matrix. Flow cytometry analyses showed that the bone marrow was composed of blood cells, including populations of monocytes, lymphocytes and granulocytes. Conclusions: Hematopoietic microenvironment engineering has great potential for hematopoietic disease therapy. The regeneration of the hematopoietic system using ASCs might be useful in the future for treating hematopoietic stem cell diseases such as leukemia as well as fibrotic bone marrow diseases.

scholarly journals Bone Marrow Regeneration Using Adipose-Derived Stem Cells

2006 ◽  
Vol 73 (1) ◽  
pp. 45-47 ◽  
Author(s):  
Rei Ogawa ◽  
Hiroshi Mizuno ◽  
Hiko Hyakusoku ◽  
Takashi Shimada
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Adipose Derived Stem Cells ◽  
Bone Marrow Regeneration

scholarly journals Human embryonic stem cell–derived hematopoietic cells are capable of engrafting primary as well as secondary fetal sheep recipients

Blood ◽  
2006 ◽  
Vol 107 (5) ◽  
pp. 2180-2183 ◽  
Author(s):  
A. Daisy Narayan ◽  
Jessica L. Chase ◽  
Rachel L. Lewis ◽  
Xinghui Tian ◽  
Dan S. Kaufman ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
Bone Marrow Cells ◽  
Hematopoietic Cells ◽  
Embryonic Stem ◽  
Xenograft Model ◽  
Fetal Sheep ◽  
Hematopoietic Stem ◽  
Hematopoietic Activity

The human/sheep xenograft model has proven valuable in assessing the in vivo hematopoietic activity of stem cells from a variety of fetal and postnatal human sources. CD34+/lineage- or CD34+/CD38- cells isolated from human embryonic stem cells (hESCs) differentiated on S17 feeder layer were transplanted by intraperitoneal injections into fetal sheep. Chimerism in primary transplants was established with polymerase chain reaction (PCR) and flow cytometry of bone marrow and peripheral blood samples. Whole bone marrow cells harvested from a primary recipient were transplanted into a secondary recipient. Chimerism was established as described before. This animal was stimulated with human GM-CSF, and an increase in human hematopoietic activity was noted by flow cytometry. Bone marrow aspirations cultured in methylcellulose generated colonies identified by PCR to be of human origin. We therefore conclude that hESCs are capable of generating hematopoietic cells that engraft primary recipients. These cells also fulfill the criteria for long-term engrafting hematopoietic stem cells as demonstrated by engraftment and differentiation in the secondary recipient.

Both alleles of PSF1 are required for maintenance of pool size of immature hematopoietic cells and acute bone marrow regeneration

Blood ◽  
2009 ◽  
Vol 113 (3) ◽  
pp. 555-562 ◽  
Author(s):  
Masaya Ueno ◽  
Machiko Itoh ◽  
Kazushi Sugihara ◽  
Masahide Asano ◽  
Nobuyuki Takakura
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Steady State ◽  
Hematopoietic Cells ◽  
Pool Size ◽  
Wild Type ◽  
Hematopoietic Stem ◽  
Molecular Stability ◽  
Evolutionarily Conserved ◽  
Bone Marrow Regeneration

Abstract Hematopoietic stem cells (HSCs) have a very low rate of cell division in the steady state; however, under conditions of hematopoietic stress, these cells can begin to proliferate at high rates, differentiate into mature hematopoietic cells, and rapidly reconstitute ablated bone marrow (BM). Previously, we isolated a novel evolutionarily conserved DNA replication factor, PSF1 (partner of SLD5-1), from an HSC-specific cDNA library. In the steady state, PSF1 is expressed predominantly in CD34+KSL (c-kit+/Sca-1+/Lineage−) cells and progenitors, whereas high levels of PSF1 expression are induced in KSL cells after BM ablation. In 1-year-old PSF1+/− mice, the pool size of stem cells and progenitors is decreased. Whereas young PSF1+/− mutant mice develop normally, are fertile, and have no obvious differences in hematopoiesis in the steady state compared with wild-type mice, intravenous injection of 5-fluorouracil (5-FU) is lethal in PSF1+/− mice, resulting from a delay in induction of HSC proliferation during ablated BM reconstitution. Overexpression studies revealed that PSF1 regulates molecular stability of other GINS components, including SLD5, PSF2, and PSF3. Our data indicate that PSF1 is required for acute proliferation of HSCs in the BM of mice.

scholarly journals Expression of CD34 marker in the adipose tissue derived stem cells

2018 ◽  
Vol 5 (1) ◽  
Author(s):  
Phuc Van Pham ◽  
Ngoc Bich Vu ◽  
Van Hong Tran
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Hematopoietic Stem Cells ◽  
Adipose Derived Stem Cells ◽  
Differentiation Potential ◽  
Hematopoietic Stem ◽  
Cd34 Expression

Introduction: Adipose-derived stem cells (ADSCs) are considered as mesenchymal stem cells (MSCs). Indeed, they display all characteristics of MSCs that compliant with the minimal criteria of MSCs suggested by Domonici et al. (2006). However, some recent studies showed that ADSCs contain the subpopulation that was positive with CD34 marker – a marker of hematopoietic stem cells. This study aimed to analyze and determine the expression of CD34 marker in ten samples of ADSCs obtained from 10 donors. Methods: All ADSC samples were isolated and expanded according to the published previous protocols. They were confirmed as the MSCs with some markers and differentiation potential, excepting the CD34 expression. Then they were cultured and analyzed the expression of CD34 by flow cytometry at passage 3, 5, 7 and 9. Results: The results showed that expression of CD34 in ADSCs was different between donors and their passages that accounted from 1.21% to 23.38%. Conclusion: These results suggested that ADSCs are not ‘truly” MSCs like MSCs from bone marrow.

scholarly journals Morphological, molecular and functional differences of adult bone marrow- and adipose-derived stem cells isolated from rats of different ages

2012 ◽  
Vol 318 (16) ◽  
pp. 2034-2048 ◽  
Author(s):  
Cristina Mantovani ◽  
Stefania Raimondo ◽  
Maryam S. Haneef ◽  
Stefano Geuna ◽  
Giorgio Terenghi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Adipose Derived Stem Cells ◽  
Adult Bone Marrow ◽  
Functional Differences ◽  
Different Ages ◽  
Adult Bone

scholarly journals ZNF521 Represses Osteoblastic Differentiation in Human Adipose-Derived Stem Cells

2018 ◽  
Vol 19 (12) ◽  
pp. 4095 ◽  
Author(s):  
Emanuela Chiarella ◽  
Annamaria Aloisio ◽  
Stefania Scicchitano ◽  
Valeria Lucchino ◽  
Ylenia Montalcini ◽  
...  
Keyword(s):  
Stem Cells ◽  
Zinc Finger Protein ◽  
Lentiviral Vector ◽  
Adipogenic Differentiation ◽  
Bone Matrix ◽  
Osteoblastic Differentiation ◽  
Adipose Derived Stem Cells ◽  
Matrix Mineralization ◽  
Calcium Deposits ◽  
Vector Transduction

Human adipose-derived stem cells (hADSCs) are multipotent mesenchymal cells that can differentiate into adipocytes, chondrocytes, and osteocytes. During osteoblastogenesis, the osteoprogenitor cells differentiate into mature osteoblasts and synthesize bone matrix components. Zinc finger protein 521 (ZNF521/Zfp521) is a transcription co-factor implicated in the regulation of hematopoietic, neural, and mesenchymal stem cells, where it has been shown to inhibit adipogenic differentiation. The present study is aimed at determining the effects of ZNF521 on the osteoblastic differentiation of hADSCs to clarify whether it can influence their osteogenic commitment. The enforced expression or silencing of ZNF521 in hADSCs was achieved by lentiviral vector transduction. Cells were cultured in a commercial osteogenic medium for up to 20 days. The ZNF521 enforced expression significantly reduced osteoblast development as assessed by the morphological and molecular criteria, resulting in reduced levels of collagen I, alkaline phosphatase, osterix, osteopontin, and calcium deposits. Conversely, ZNF521 silencing, in response to osteoblastic stimuli, induced a significant increase in early molecular markers of osteogenesis and, at later stages, a remarkable enhancement of matrix mineralization. Together with our previous findings, these results show that ZNF521 inhibits both adipocytic and osteoblastic maturation in hADSCs and suggest that its expression may contribute to maintaining the immature properties of hADSCs.

An electrochemically deposited collagen wound matrix combined with adipose-derived stem cells improves cutaneous wound healing in a mouse model of type 2 diabetes

2018 ◽  
Vol 33 (4) ◽  
pp. 553-565 ◽  
Author(s):  
Nicole Edwards ◽  
Denis Feliers ◽  
Qingwei Zhao ◽  
Randolph Stone ◽  
Robert Christy ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Stem Cells ◽  
Mouse Model ◽  
Chronic Wounds ◽  
High Porosity ◽  
Adipose Derived Stem Cells ◽  
Clinical Issue ◽  
Wound Matrix ◽  
Electrochemically Deposited

Chronic wounds complicated by diabetes are a significant clinical issue, and their occurrence is expected to continue to rise due to an increased prevalence of diabetes mellitus, especially type 2 diabetes. Diabetic wounds frequently lead to nonhealing ulcers, and often eventually result in limb amputation due to the high risk of infection of the chronic wound. Here, we present a tissue-engineered treatment that combines a novel electrochemically deposited collagen wound matrix and human adipose-derived stem cells. The matrix fabrication process is optimized for voltage and time, and the final collagen biomaterial is thoroughly characterized. This collagen material possesses high tensile strength, high porosity, and excellent biocompatibility and cellular proliferation capabilities. Human adipose-derived stem cells were seeded onto the collagen wound matrix and this construct is investigated in a full thickness excisional wound in a mouse model of type 2 diabetes. This novel treatment is shown to stimulate excellent healing and tissue regeneration, resulting in increased granulation tissue formation, epidermal thickness, and overall higher quality tissue reformation. Both the collagen wound matrix alone and collagen wound matrix in combination with adipose derived stem cells appeared to be excellent treatments for diabetic skin wounds, and in the future can also be optimized to treat other injuries such as burns, blast injuries, surgical incisions, and other traumatic injuries.

scholarly journals Primitive hematopoietic cells in murine bone marrow express the CD34 antigen

Blood ◽  
1996 ◽  
Vol 88 (10) ◽  
pp. 3774-3784 ◽  
Author(s):  
F Morel ◽  
SJ Szilvassy ◽  
M Travis ◽  
B Chen ◽  
A Galy
Keyword(s):  
Stem Cells ◽  
Monoclonal Antibody ◽  
Bone Marrow ◽  
Progenitor Cells ◽  
Significant Proportion ◽  
Hematopoietic Cells ◽  
Full Detail ◽  
Hematopoietic Stem ◽  
Cd34 Antigen

The CD34 antigen is expressed on most, if not all, human hematopoietic stem cells (HSCs) and hematopoietic progenitor cells, and its use for the enrichment of HSCs with repopulating potential is well established. However, despite homology between human and murine CD34, its expression on subsets of primitive murine hematopoietic cells has not been examined in full detail. To address this issue, we used a novel monoclonal antibody against murine CD34 (RAM34) to fractionate bone marrow (BM) cells that were then assayed in vitro and in vivo with respect to differing functional properties. A total of 4% to 17% of murine BM cells expressed CD34 at intermediate to high levels, representing a marked improvement over the resolution obtained with previously described polyclonal anti-CD34 antibodies. Sixty percent of CD34+ BM cells lacked lineage (Lin) markers expressed on mature lymphoid or myeloid cells. Eighty-five percent of Sca-1+Thy-1(10)Lin- /10 cells that are highly enriched in HSCs expressed intermediate, but not high, levels of CD34 antigen. The remainder of these phenotypically defined stem cells were CD34-. In vitro colony-forming cells, day-8 and -12 spleen colony-forming units (CFU-S), primitive progenitors able to differentiate into B lymphocytes in vitro or into T lymphocytes in SCID mice, and stem cells with radioprotective and competitive long-term repopulating activity were all markedly enriched in the CD34+ fraction after single-parameter cell sorting. In contrast, CD34-BM cells were depleted of such activities at the cell doses tested and were capable of only short-term B-cell production in vitro. The results indicate that a significant proportion of murine HSCs and multilineage progenitor cells express detectable levels of CD34, and that the RAM34 monoclonal antibody is a useful tool to subset primitive murine hematopoietic cells. These findings should facilitate more direct comparisons of the biology of CD34+ murine and human stem and progenitor cells.

scholarly journals Syngeneic transplantation of hematopoietic stem cells that are genetically modified to express factor VIII in platelets restores hemostasis to hemophilia A mice with preexisting FVIII immunity

Blood ◽  
2008 ◽  
Vol 112 (7) ◽  
pp. 2713-2721 ◽  
Author(s):  
Qizhen Shi ◽  
Scot A. Fahs ◽  
David A. Wilcox ◽  
Erin L. Kuether ◽  
Patricia A. Morateck ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Transgenic Mice ◽  
Hematopoietic Stem Cells ◽  
Factor Viii ◽  
Hemophilia A ◽  
Genetically Modified ◽  
Hematopoietic Stem ◽  
Inhibitory Antibodies ◽  
Sublethal Irradiation

Abstract Although genetic induction of factor VIII (FVIII) expression in platelets can restore hemostasis in hemophilia A mice, this approach has not been studied in the clinical setting of preexisting FVIII inhibitory antibodies to determine whether such antibodies would affect therapeutic engraftment. We generated a line of transgenic mice (2bF8) that express FVIII only in platelets using the platelet-specific αIIb promoter and bred this 2bF8 transgene into a FVIIInull background. Bone marrow (BM) from heterozygous 2bF8 transgenic (2bF8tg+/−) mice was transplanted into immunized FVIIInull mice after lethal or sublethal irradiation. After BM reconstitution, 85% of recipients survived tail clipping when the 1100-cGy (myeloablative) regimen was used, 85.7% of recipients survived when 660-cGy (nonmyeloablative) regimens were used, and 60% of recipients survived when the recipients were conditioned with 440 cGy. Our further studies showed that transplantation with 1% to 5% 2bF8tg+/− BM cells still improved hemostasis in hemophilia A mice with inhibitors. These results demonstrate that the presence of FVIII-specific immunity in recipients does not negate engraftment of 2bF8 genetically modified hematopoietic stem cells, and transplantation of these hematopoietic stem cells can efficiently restore hemostasis to hemophilic mice with preexisting inhibitory antibodies under either myeloablative or nonmyeloablative regimens.

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