Bone Marrow Cells ; cytology ; physiology ; Humans ; Immunity ; Mesenchymal Stromal Cells ; cytology ; physiology

Bone marrow mensenchymal stem cells (BM-MSCs) are capable of supporting the survival, differentiation and migration of hematopoietic stem cell, and have a profound application prospect in transplantation and treatment of graft-versus-host disease (GVHD). This review aims to illustrate the interaction between BM-MSCs and other intra-bone marrow cells, including hematopoietic stem cells, endothelial cells and osteoblasts. The investigation of their regulating mechanism will help better understanding of the BM-MSCs' role in hematopoiesis.
Bone Marrow Cells ; cytology ; Cell Communication ; physiology ; Endothelial Cells ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology

OBJECTIVETo investigate the feasibility of using marrow stromal osteoblast-cancellous bone matrix compound artificial bone (MCCAB) as tissue-engineered bone, the osteogenesis of MCCAB in the cranial defect was observed in the experiment.

METHODSThe in vitro cultivated and induced marrow stromal cells of adult New Zealand rabbits were seeded into the alginate-cancellous bone matrix to form MCCAB. The MCCAB was then implanted into the cranial defect for 4 to 8 weeks. The cancellous bone matrix (CBM) alone or the marrow stromal osteoblasts (MSOs) alone was implanted as the control. The effectiveness of bone formation was assessed by histological and roentgenographic analysis.

RESULTSThe osteogenesis of MCCAB was better than CBM or MSOs and superior to the blank group.

CONCLUSIONMCCAB can effectively repair cranial defect. It could be used clinically to restore large bone defects.

Animals ; Bone Marrow Cells ; cytology ; physiology ; Bone Matrix ; cytology ; Cells, Cultured ; Feasibility Studies ; Male ; Osteoblasts ; cytology ; physiology ; Rabbits ; Skull ; abnormalities ; Stromal Cells ; cytology ; physiology

It was recently discovered that a subset of osteoblasts functions as a key component of the hematopoietic stem cells (HSC) niche in vivo, controlling HSC self-renewal and multi-lineage differentiation. Disruption of Smad4 gene specifically in osteoblasts leads to a remarkable decrease of osteoblast number and endosteal surface area. In order to elucidate if the osteoblast loss has any effect on hematopoietic activity, the bone marrow (BM) and extramedullary hematopoiesis in the osteoblast-specific Smad4 knockout mice were systematically analyzed, the proportions of mature hematocytes in BM, liver and spleen were detected by flow cytometry, the hematopoietic progenitor number in different stages was measured by colong-forming assay, CFU-S and analysis of LSK cells. The results indicated that the conditional mutant mice demonstrated normal BM hematopoiesis without sign of extramedullary hematopoiesis. Furthermore, the proportion of hematopoietic progenitor cells was normal, while cell number/body weight of the conditional knockout mice increased. It is concluded that hematopoiesis is normally maintained in osteoblast-specific Smad4 knockout mice, and osteoblast loss does not of necessity result in the decrease in BM hematopoiesis.
Animals ; Bone Marrow Cells ; cytology ; Hematopoiesis ; Hematopoietic Stem Cells ; cytology ; physiology ; Mice ; Mice, Knockout ; Osteoblasts ; cytology ; physiology ; Smad4 Protein ; genetics

OBJECTIVETo review the progress of cardiac differentiation and electrophysiological characteristics of bone marrow mesenchymal stem cells.

DATA SOURCESThe databases of PubMed, Springer Link, Science Direct and CNKI were retrieved for papers published from January 2000 to January 2012 with the key words of "bone marrow mesenchymal stem cells, cardiac or heart, electrophysiology or electrophysiological characteristics".

STUDY SELECTIONThe articles concerned cardiac differentiation and electrophysiological characteristics of bone marrow mesenchymal stem cells were collected. After excluding papers that study purposes are not coincident with this review or contents duplicated, 56 papers were internalized at last.

RESULTSFor the treatment of myocardial infarction and myocardiac disease, the therapeutic effects of transplantation of bone marrow mesenchymal stem cells which have the ability to develop into functional myocardial cells by lots of methods have been proved by many researches. But the arrhythmogenic effect on ventricles after transplantation of bone marrow mesenchymal stem cells derived myocardial cells is still controversial in animal models. Certainly, the low differentiation efficiency and heterogeneous development of electrical function could be the most important risk for proarrhythmia.

CONCLUSIONMany studies of cardiac differentiation of bone marrow mesenchymal stem cells have paid attention to improve the cardiac differentiation rate, and the electrophysiology characteristics of the differentiated cells should be concerned for the risk for proarrhythmia as well.

Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Electrophysiology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Myocardial Infarction ; therapy ; Myocytes, Cardiac ; cytology ; physiology

Matrix metalloproteinases (MMPs) are endocellular proteolytic enzymes. They are so named because they need Ca2+, Zn2+ and other metal ions as their cofactors. MMPs play an important biological role in regulating the formation, remodeling and degradation of extracellular matrix and participate in various physiological and pathological processes of cells. Bone marrow-derived mesenchymal stem cells (BMSCs) are a kind of pluripotent stem cell which has the ability to self-renew and differentiate into functional cells. Meanwhile, they can respond to the damage signals and migrate to injured site for tissue repair and regeneration. MMPs and their inhibitors TIMPs affect the differentiation and migration of BMSCs. This article reviews the roles of MMPs in differentiation and migration of BMSCs.
Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cell Movement ; physiology ; Humans ; Matrix Metalloproteinases ; physiology ; Mesenchymal Stromal Cells ; cytology

BACKGROUNDTreatments to regenerate different tissue involving the transplantation of bone marrow derived mesenchymal precursor cells are anticipated. Using an alternative methods, in vitro organotypic slice culture method, would be useful to transplant cells and assessing the effects. This study was to determine the possibility of differentiating human bone marrow precursor cells into cells of the neuronal lineage by transplanting into canine spinal cord organotypic slice cultures.

METHODSBone marrow aspirates were obtained from posterior superior iliac spine (PSIS) of patients that had undergone spinal fusion due to a degenerative spinal disorder. For cell imaging, mesenchymal precursor cells (MPCs) were pre-stained with PKH-26 just before transplantation to canine spinal cord slices. Canine spinal cord tissues were obtained from three adult beagle dogs. Spinal cords were cut into transverse slices of 1 mm using tissue chopper. Two slices were transferred into 6-well plate containing 3 ml DMEM with antibiotics. Prepared MPCs (1×10(4)) were transplanted into spinal cord slices. On days 0, 3, 7, 14, MPCs were observed for morphological changes and expression of neuronal markers through immunofluorescence and reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe morphological study showed: spherical cells in the control and experiment groups on day 0; and on day 3, cells in the control group had one or two thick, short processes and ones in the experiment group had three or four thin, long processes. On day 7, these variously-sized processes contacted each other in the experiment group, but showed typical spindle-shaped cells in the control group. Immunofluorescence showed that PKH-26(+) MPCs stained positive for NeuN(+) and GFAP(+) in experimental group only. Also RT-PCR showed weak expression of β-tubulin III and GFAP.

CONCLUSIONSHuman bone marrow mesenchymal precursor cells (hMPCs) have the potential to differentiate into the neuronal like cells in this canine spinal cord organotypic slice culture model. Furthermore, these findings suggested the possibility that these cells can be utilized to treat patients with spinal cord injuries.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Dogs ; Humans ; Mesenchymal Stromal Cells ; cytology ; Spinal Cord ; cytology

OBJECTIVETo induce the differentiation of bone marrow stromall cells (BMSCs) isolated from Beagles into osteoblasts in vitro and identify the osteogenic potential and bioactivity of the BMSCs.

METHODSPrimary cultured BMSCs isolated from Beagles were subcultured in mineralization medium to induce their differentiation into osteoblasts, whose morphological characteristics and proliferation status were observed by phase-contrast microscope. The osteogenic activity of the cells was evaluated with von Kossa staining of the mineralized nodules and determination of the alkaline phosphatase activity.

RESULTBMSCs cultured in vitro showed obvious osteogenic capacity in DMEM. Von Kossa staining of the mineralized nodules and alkaline phosphatase detection of the passaged cells both yielded positive results.

CONCLUSIONBMSCs cultured in vitro contain osteogenic precursor cells, and the passaged cells possess osteogenic potential.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Dogs ; Male ; Osteoblasts ; cytology ; Osteogenesis ; Stromal Cells ; cytology

A systematic method of isolating and culturing human bone mesenchymal stem cells (hMSCs), and inducing them to differentiate into neuron-like cells in vitro was established. The hMSCs were isolated from bone marrow with the lymphocyte-separating medium, cultured and expanded in vitro, and induced after addition of compound neuro-revulsants. The morphological changes of hMSCs were observed, and the expression of surface markers in induced hMSCs was immunocytochemically identified during induction period. The hMSCs could be separated, cultured and expanded in vitro. After induction by compound neuro-revulsants for 48 h, the changes of neuron-like cells, such as cellular shrinkage and neurite growth, were observed in some cells. The immunochemical staining revealed nestin (+) or NF (+), and GFAP (-). It was concluded that hMSCs were successfully cultured and induced to differentiate into neuron-like cells.
Bone Marrow Cells/*cytology ; Cell Culture Techniques ; Cell Differentiation/*physiology ; Cells, Cultured ; Mesenchymal Stem Cells/*cytology ; Neurons/*cytology

To investigate the effects of human mesenchymal stem cells (MSC) and human fibroblastoid cell line (HFCL) as feeder layer on expansion of umbilical cord blood CD34(+) cells in vitro, (60)Co gamma-ray irradiated MSC and HFCL were used as feeder layer to expand cord blood CD34(+) cells in culture. The efficiencies of MSC and HFCL on expansion of CD34(+) cells in culture with or without cytokines were compared. The results showed that no matter whether cytokines (rhFL, rhSCF, rhTPO) were added, the proliferation of nucleated cells after expansion for 12 days in HFCL group was statistically higher than that in MSC group, i.e. with cytokines (9797 +/- 361)% vs (7061 +/- 418)%; without cytokines (5305 +/- 354)% vs (1992 +/- 247)%, when the cell numbers at day 0 was accounted as 100%), P < 0.01. The proliferation of propagated CD34(+) cells between MSC group and HFCL without addition of cytokines was not statistically different (820 +/- 191)% vs (825 +/- 305)%, P > 0.05. However, in the presence of cytokines, the propagating rate of MSC group was lower than that of HFCL group (939 +/- 212)% vs (1617 +/- 222)%, P < 0.01. MSC was better than HFCL in maintaining the LTC-IC of UCB CD34(+) cells, i.e. the number of CFU-GM colonies in the fifth week was (129.95 +/- 8.73) /10(5) seeded cells vs (89.81 +/- 10.29) colonies/10(5) cells, P < 0.05; with addition of cytokines, the effect was more obvious, i.e. the number of CFU-GM colonies in the fifth week (192.93 +/- 4.95)/10(5) seeded cells vs (90.47 +/- 14.28) colonies/10(5) seeded cells, P < 0.01. MSC mixed with a certain proportion of HFCL facilitated maintaining the LTC-IC of UCB CD34(+) cells. When the proportion was 4:1, the number of CFU-GM colonies was the highest (186.89 +/- 11.11)/10(5) seeded cells, which was higher than that of both 3:2 group [(138.92 +/- 14.84) colonies/10(5) seeded cells] and MSC only group, i.e. (64.63 +/- 6.11) colonies/10(5) seeded cells, both P < 0.01. It is concluded that HFCL is better than MSC in maintaining the expansion of CD34(+) cells and cytokines can enhance this effect, while MSC are stronger than HFCL in maintaining the LTC-IC of UCB CD34(+) cells in vitro. MSC with addition of a certain proportion of HFCL can significantly enhance the efficiency of CD34(+) cell expansion.
Antigens, CD34 ; analysis ; Bone Marrow Cells ; cytology ; physiology ; Cell Line ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Fetal Blood ; cytology ; Fibroblasts ; cytology ; physiology ; Humans ; Mesenchymal Stromal Cells ; cytology ; physiology