Megakaryocytic Progenitors in Long Term Cultures from MDS Patients.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4924-4924
Author(s):  
Maria Juliana Majado ◽  
María I. Macizo ◽  
Consuelo González-García ◽  
Eduardo Salido ◽  
José A. Molina ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Culture Medium ◽  
Horse Serum ◽  
Calf Serum ◽  
Methyl Cellulose ◽  
Refractory Anemia ◽  
Foetal Calf Serum ◽  
Cell Counts ◽  
Control Bone

Abstract Introduction: bone marrow long-term cultures (LTC) have been reported to produce very poor adherent stromal layer in MDS, but there are not many studies about the behaviour of the progenitor cells in supernatant culture medium. In this study we report the results of megakaryocytic progenitors (CFU-MK) in the supernatant of LTC of patients with refractory anemia (RA). Material and method: LTCs were performed to 11 RA patients and to 13 normal multiorgan donors as control. Bone marrow red cells were sedimented with 1% methyl-cellulose and seeded for LTC in 10ml flasks (Nalge Nunc International), with Iscover medium supplemented with horse serum, foetal calf serum, hydrocortisone and carbonic air, at a final concentration of 1x106 per ml. Flasks were placed in an incubator for 8 weeks. Half culture medium volume was renovated weekly, cell counts and assays of CFU-MK (Megacult C, Stem Cell Technologies), were performed. CFU-MK colonies were separated in three groups, containing 3–20 cells, 21–50 cells and more than 50 cells. Student t-test was used for statistical comparisons. Results: Are in the following table, expressed as mean + standard desviation. After the third week no colony growth was observed in normal such as in MDS. Growth of CFU-MK colonies containing more than 50 cells was higher in basal control bone marrow, no statistical differences were found in the rest of the cultures. Results CFU-MK(3–30 Cells) CFU-MK(20–50 Cells) CFU-MK (>50 Cells) CFU-MK total w: week Control basal 15.75±15.79 3.33±5.42 4.88±7.44 22.42±26.43 RA basal 7.85±8.66 0.80±1.01 0.70±0.98 9.45±9.46 t-St ns ns 0.047 ns Control w 1 3.54±4.42 0.7±1.44 0.74±1.48 4.98±6.62 RA w 1 3.29±4.52 0.29±0.93 0.25±0.67 3.82±5.65 t-St ns ns ns ns Control w 2 0.55±1.14 0.07±0.24 0.05±0.25 0.67±1.43 RA w 2 0.18±0.32 0.00±0.00 0.04±0.13 0.21±0.38 t-St ns ns ns ns Control w 3 0.18±0.53 0.00±0.00 0.00±0.00 0.18±0.53 RA w 3 0.00±0.21 0.00±0.00 0.00±0.00 0.00±0.21 t-St ns ns Discussion: No important difference was found in LTC supernatant CFU-MK in our RA patients, and this supports the idea that the stromal damage is more important, in the pathophysyology of MDS, than that of the stem cells.

Immunocytochemistrical Studies on Cytoskeletal Change in Cultured Cardiomyocyte of Tilapia

2000 ◽  
Vol 6 (S2) ◽  
pp. 888-889
Author(s):  
L. C. Tung
Keyword(s):  
Culture Medium ◽  
Cultured Cells ◽  
Calf Serum ◽  
Amino Acid Mixture ◽  
Penicillin G ◽  
Acid Mixture ◽  
Minimal Essential Medium ◽  
Cultured Fish ◽  
Cytoskeletal Change

In the present study, the myofibril regeneration in the long-term cultured fish cardiomyocytes was studied with immunocytochemistry.Adult Tilapia heart was dissociated into a single-cell suspension with collagenase and protease-minced tissue method. The culture medium was Eagle's minimal essential medium (MEM) with Earle's salts, supplemented with 10% fetal calf serum, 1 x nonessential amino acid mixture, 100 IU/ml penicillin G, and 100 μg/ml streptomycin. The cultured cells were grown in a humidified CO2 incubator at 28°Cand in a medium without glutamine for eliminating fibroblast contamination. In the initial 24 h culture, the elongated-shape cells gradually shortened from their both ends and rounded up. Over 5 to 6 days postcultivation, the cells attached to the bottom of the culture flask and began to protrude pseudopodia. The cells could not be subcultured and also proliferated indefinitely. The life span of cells in culture was 30 to 60 days.

Different optimal PHA concentrations for stimulation of Chinese hamster lymphocytes in cultures supplemented with foetal calf serum or horse serum

1982 ◽  
Vol 50 (1) ◽  
pp. 11-15
Author(s):  
B. De Jong ◽  
G.J.P.A. Anders ◽  
I.H. Van Der Meer ◽  
J. Zijlstra ◽  
V.J.S. Idenburg
Keyword(s):  
Horse Serum ◽  
Calf Serum ◽  
Chinese Hamster ◽  
Foetal Calf Serum ◽  
Stimulation Of

scholarly journals Immunoreactive insulin from mouse brain cells in culture and whole rat brain

1984 ◽  
Vol 218 (1) ◽  
pp. 19-27 ◽  
Author(s):  
N P Birch ◽  
D L Christie ◽  
A G C Renwick
Keyword(s):  
Rat Brain ◽  
Mouse Brain ◽  
Culture Medium ◽  
Calf Serum ◽  
Brain Cells ◽  
Immunoreactive Insulin ◽  
Foetal Calf Serum ◽  
Cells In Culture ◽  
Pancreatic Insulin ◽  
Contrast Microscopy

Foetal mouse brain cells were cultured as described previously [Sotelo, Gibbs, Gajdusek, Toh & Wurth (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 653-657] without added insulin and without foetal calf serum after 12 days in culture. Examination by phase-contrast microscopy showed that these modifications did not appear to affect growth and development of the cells adversely. Silver impregnation of the cultures and indirect immunofluorescence following reaction with tetanus toxin showed that a high proportion of the cells resembled neurones. Analysis of concentrated culture medium by radioimmunoassay and high-pressure liquid chromatography (h.p.l.c.) revealed that the cells produced two main forms of immunoreactive insulin which differed from authentic pancreatic insulin in retention time. Immunoreactive somatostatin was also produced in culture and this was resolved into at least three forms by h.p.l.c. Immunoreactive insulin was also extracted from whole rat brain by using two published procedures. The method of Havrankova, Schmechel, Roth & Brownstein [Proc. Natl. Acad. Sci. U.S.A. (1978) 75, 5737-5741] consistently gave greater yields of insulin than did that of Eng & Yalow [Diabetes (1980) 29, 105-109] and the concentration was about three times that of plasma. The extracted insulin was further characterized by h.p.l.c. in each case and was found to behave like authentic pancreatic insulin. The production of insulin and somatostatin by foetal mouse brain cells in culture suggests that they may be a useful model system for studies of neuropeptide biosynthesis.

Suppression of Cyclin D 1 (CD1) by on 01910.Na Is Associated with Decreased Survival or Trisomy 8 Myelodysplastic Bone Marrow: A Potential Targetted Therapy for Trisomy 8 MDS.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1651-1651
Author(s):  
Aarthi Shenoy ◽  
Loretta Pfannes ◽  
Francois Wilhelm ◽  
Manoj Maniar ◽  
Neal Young ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cyclin D1 ◽  
Cultured Cells ◽  
Bone Marrow Failure ◽  
Refractory Anemia ◽  
Trisomy 8 ◽  
Monosomy 7 ◽  
Diploid Cells

Abstract CD34 positive cells from patients with trisomy 8 myelodysplastic syndrome (MDS) have pronounced expression of early apoptotic markers compared to normal hematopoietic cells. However, trisomy 8 clones persist in patients with bone marrow failure and expand following immunosuppression (Sloand EM et al; Blood2005; 106(3):841). We have demonstrated up-regulation of c-myc, survivin, and CD1 in CD34 cells of patients with trisomy 8 (Sloand et al; Blood2007; 109(6):2399). Employing siRNA mediated knockdown of the anti-apoptotic protein survivin, we demonstrated a decrease in trisomy 8 cell growth and postulated that increased Cyclin D1 caused the upregulation of survivin resulting in resistance of these cells to apoptosis. Using fluorescent in situ hybridization (FISH) we showed that the novel styryl sulfone, ON 01910.Na (Vedula MS et al; European Journal of Medicinal Chemistry2003;38:811), inhibits cyclin D1 accumulation and is selectively toxic to trisomy 8 cells while promoting maturation of diploid cells. Flow cytometry of cultured cells demonstrated increased proportions of mature CD15 positive myeloid cells and decreased number of immature CD33+ cells or CD34+ blasts (Sloand EM et al; Blood2007;110:822). These encouraging in vitro data led to a phase I/II trial of ON 01910.Na in MDS patients with refractory anemia with excess blasts who had IPSS =/> int-2. This study was designed to assess the safety, and activity of escalating doses of ON 01910.Na (800 mg/m2/day × 3 days, 800 mg/m2/day × 5 days, 1500 mg/m2/day × 5 days, 1800 mg/m2/day × 5 days every 2 weeks) in MDS patients. To date five MDS patients have been treated with ON 01910.Na for 4 to 16 weeks in the first two dose cohorts. Two patients had isolated trisomy 8, two had complex cytogenetic abnormalities including trisomy 8 in all aneuploid cells, and one had monosomy 7. Three and five day infusions were well tolerated. Pharmakokinetic analysis showed that the half life of the drug is 1.3 ± 0.5 hours without signs of drug accumulation. Four of five patients demonstrated a rapid and significant decrease in the number of peripheral blasts and aneuploid cells after 4 weeks of therapy (see below), concomitantly with increases in neutrophil and/or platelet counts in four patients. All four patients exhibiting a biological effect of drug treatment had trisomy 8 in their aneuploid clone prior to therapy. One monosomy 7 patient, previously refractory to EPO became responsive to Darbopoietin and another trisomy 8 patient became platelet-transfusion independent. In this early safety trial, ON 01910.Na demonstrates efficacy at early timepoints with respect to improved cytopenias and decreased blast counts. Continued enrollment and long term follow-up will further detail clinical efficacy and impact on the long term prognosis of high risk MDS patients treat with this drug. Figure Figure

Spectral Karyotyping (SKY) Reveals a New Subset of MDS Patients with Clonal Chromosomal Abnormalities Not Detected by G-Banding Analysis

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1718-1718
Author(s):  
Fabio Morato Oliveira ◽  
Maria do Carmo Favarin ◽  
Rodrigo T Calado ◽  
Ana Paula N Rodrigues Alves ◽  
Cassia Godoi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cytogenetic Analysis ◽  
Chromosomal Abnormalities ◽  
Bone Marrow Cells ◽  
Conflicts Of Interest ◽  
Calf Serum ◽  
Fluorescent Labeling ◽  
Refractory Anemia ◽  
Spectral Karyotyping ◽  
Banding Analysis

Abstract Abstract 1718 Cytogenetic findings in bone marrow cells of MDS patients are essential for a correct diagnosis and classification of the disease and constitute one of the most important independent prognostic factors. The classical cytogenetic analysis, however, often cannot be fully resolved by G-banding because of the presence of marker chromosomes, rings or unidentified material attached to chromosomes. Spectral karyotyping (SKY) has proven to be an important tool for the interpretation of complex karyotypes or identification of suitable abnormalities in hematological malignancies. By using SKY analysis in combination with G-banding were identified new clonal chromosomal abnormalities “masked” by the limited resolution of classical cytogenetic. As a consequence changes in IPSS score were observed. Bone marrow samples of 46 (forty-six) MDS patients were incubated in RPMI 1640 with 20% fetal calf serum for 72h at 37°C. Chromosome preparations were obtained by using standard procedures and the subsequent cytogenetic analysis and interpretation were made according to ISCN 2009. The patients studied were classified as refractory anemia (RA) and refractory anemia with ringed sideroblast (RARS), with less than 5% blast. Slides for SKY were prepared by using the same fixed chromosome preparations, stored at −20°C, as employed for G-banding analysis. Chromosome labeling was performed with the SKY fluorescent labeling kit (Applied Spectral Imaging, Migdal HaEmek, Israel) according to the manufacturer's protocol. A minimum of twenty metaphases were analyzed using the SkyView 5.5 software (ASI, Carlsbad, CA, USA). In a group of 46 subjects studied, the cytogenetic analysis (G-banding) showed chromosomal aberrations in 13 patients (54.2%) and normal karyotype was observed in 11 subjects (45.8%). The abnormalities observed were dup(1)(q21q32), inv(3)(q21q26), t(3;3)(q21;q26), +4, del(5)(q31), −7, del(7)(q22q36), +8, add(17)(p12), +i(17)(q10), del(20)(q11). The group with normal cytogenetic, SKY analysis revealed “masked” chromosomal abnormalities in 6 patients, being t(7;9)(q36;q34), ins(1;6)(q21;?), t(11;12)(p15;q24.1), ins(3;5)(p21;?), t(8;16)(q23;?) and ins(6;11)(q21;?). Among 13 cases studied with previous chromosomal abnormalities by G-banding analysis, SKY identified additional abnormalities in 8 patients. Some abnormalities found include t(6;9)(q27;q22), t(12;17)(p13;p12) and t(8;11)(p12;q12). For both groups with normal and altered karyotypes, the profile of masked chromosomal abnormalities seen were insertions and translocations involving small segments of chromosomes. In the majority of the cases the frequency of abnormal clones was less than 50%. However, in all patients the abnormalities identified by SKY were classified as clonal. All abnormalities identified were confirmed by FISH, by using a set of probes. SKY analysis has proved to be a promising and reliable method for identification of additional and complex chromosomal abnormalities usually present in a great number of human neoplasias. The contribution for the prognostic information of these new chromosomal abnormalities identified beyond the limited resolution of G-banding in MDS will require a detail analysis of the patients' evolution. Financial Support: FAPESP (Proc. 07/52462-7) Disclosures: No relevant conflicts of interest to declare.

scholarly journals Latexin Inhibition Mitigates Irradiation Induced Hematopoietic Injury

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 2562-2562
Author(s):  
Cuiping Zhang ◽  
Xiaojing Cui ◽  
Ying Liang
Keyword(s):  
Bone Marrow ◽  
Cell Death ◽  
Progenitor Cells ◽  
Knockout Mice ◽  
Lethal Dose ◽  
Negative Regulator ◽  
Cell Counts ◽  
Survival Pathway ◽  
Radiation Induced

Abstract Radiation-associated bone marrow (BM) injury is one of the most serious limiting factors of radiotherapy. Radiation-induced hematopoietic injury, no matter how transient or long lasting, can ultimately impair HSC function and decrease the HSC reserve, leading to increased risk for the development of BM failure or cancer. However, molecular mechanisms underlying radiation-induced HSC functional decline are largely unknown. We previously identified a stem cell regulatory gene, latexin (Lxn), as a novel negative regulator of HSCs in mice. HSCs in Lxn knockout mice (Lxn-/-) had increased self-renewal and survival. In our new findings, we surprisingly found that Lxn-/- mice had the significant survival advantages under lethal dose of total body irradiation (TBI). We further found that HSCs and hematopoietic progenitor cells (HPCs), measured by immunophenotypes and colony assay, recovered much faster in Lxn-/- mice than wild-type mice (WT) within one month after sub-lethal dose of TBI. The better preserved HSC/HPC pool was due to the decreased apoptosis in which the percentage of Annexin V + PI- apoptotic HSCs/HPCs cells was significantly lower in Lxn-/- mice than WT mice. These data suggest that Lxn inactivation protects HSCs and HPCs from radiation-induced cell death, thus mitigating acute hematopoietic suppression and conferring a survival advantage. To determine the long-term effect of TBI on Lxn-/- HSCs, we performed limiting dilution competitive repopulation unit assay (CRU), and found that Lxn-/- CRU was significantly higher than WT CRU. Moreover, we performed serial transplantation experiment, and found that Lxn-/- HSC continuously regenerated blood and bone marrow cells even at the 4th round of transplantation whereas WT HSCs were exhausted. These data provide robust evidence that Lxn inactivation protects functional long-term HSCs from radiation-induced injury. Radiation can increase the risk of hematological malignancy later in the life. We thus maintained a group of mice that were subject to either a single dose of 6.5Gy TBI or split low doses of TBI (2 Gy daily for 6 days), and monitored their gross condition and blood cell counts for 20 months. At 20 month post-radiation, we performed bone marrow analysis and histopathology analysis. We found that Lxn-/- mice did not spontaneously develop hematopoietic malignancies, their bone marrow HSCs/HPCs had normal population size, and bone marrow had normal histopathology. These data suggest that Lxn inactivation mitigates radiation-induced short-term myelosuppression and long-term HSC functional impairment without induction of hematologic malignancy. At the molecular level, we previously reported that Lxn sensitized leukemogenic cells to gamma-irradiation-induced cell-cycle arrest and cell death through Rps3 pathway, and Rps3 was a binding protein of Lxn. Rps3 has been shown to be involved in the NFkB pathway. We found that Rps3 bound Lxn in primary hematopoietic stem and progenitor cells (HSPCs) using Co-IP assay. Lxn-/- HSPCs had the increased expression of Rps3 and NFkB p65 before or post-irradiation. Knockdown of Rps3 in Lxn-/- HSPCs decreased NFkB p65 and increased radiation-induced apoptosis. Moreover, when Lxn-/- HSPCs were treated with NFkB p65 specific inhibitor, the similar phenotypes were also shown, suggesting that Lxn functions through Rps3-NFkB-mediated pro-survival pathway in primary HSPCs. We are currently proving this molecular pathway using the in vivo model by crossing p65 knockout mice with Lxn-/- mice. In conclusion, latexin inhibition mitigates irradiation induced hematopoietic injury via Rps3-NFkB-mediated pro-survival pathway. Disclosures No relevant conflicts of interest to declare.

scholarly journals Proliferation of myeloid progenitor cells in human long-term bone marrow cultures is stimulated by interleukin-1 beta

Blood ◽  
1988 ◽  
Vol 72 (4) ◽  
pp. 1242-1247 ◽  
Author(s):  
WE Fibbe ◽  
HM Goselink ◽  
G Van Eeden ◽  
J Van Damme ◽  
A Billiau ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Culture Medium ◽  
Interleukin 1 ◽  
Hematopoietic Progenitor ◽  
Bone Marrow Cultures ◽  
Fresh Culture Medium ◽  
Fresh Culture ◽  
Number Of Cells

Abstract To study the effect of interleukin-1 (IL-1) beta on the proliferation of hematopoietic progenitor cells (HPC) in long-term bone marrow cultures (LTBMC), stromal cell layers were established from normal human bone marrow. Autologous cryopreserved mononuclear phagocyte- and T-lymphocyte-depleted bone marrow cells were reinoculated on the stromal layers in fresh culture medium, with or without the addition of human IL-1 beta (30 U/mL). Once a week, half of the culture supernatant was replaced with fresh culture medium with or without IL-1, and all nonadherent cells were returned to the flasks. At weekly intervals during a period of 5 weeks, one culture was sacrificed to determine the total number of cells and hematopoietic progenitor cells, present in the adherent and the nonadherent cell fractions. In IL-1-stimulated cultures, the number of cells recovered during a period of 5 weeks exceeded the number of cells in unstimulated control cultures by 1.5 times. This difference was attributed to a twofold increase in the number of adherent cells. The number of HPC recovered from IL-1- stimulated cultures was not different from that recovered from controls. The levels of colony-stimulating activity (CSA) in supernatants from IL-1-stimulated cultures were significantly higher than those in supernatants from control cultures. These results indicate that IL-1 enhances the recovery of cells in LTBMC by stimulating the proliferation of HPC with the concurrent release of CSA from stromal cells, without diminishing the number of HPC.

scholarly journals In vitro establishment and characterization of a human megakaryoblastic cell line

Blood ◽  
1990 ◽  
Vol 75 (6) ◽  
pp. 1252-1261 ◽  
Author(s):  
DA Fugman ◽  
DP Witte ◽  
CL Jones ◽  
BJ Aronow ◽  
MA Lieberman
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Cell Line ◽  
Solid Tumor ◽  
Calf Serum ◽  
Culture Conditions ◽  
Platelet Factor ◽  
Defined Culture

A human megakaryoblastic cell line, designated CHRF-288–11, has been established in vitro through the use of adherent stromal cells in long- term human bone marrow culture. Long-term bone marrow cultures were required for the initial adaptation of the megakaryoblastic cells to culture conditions; however, once adapted, the cells were weaned from the stromal layer until they proliferated in the complete absence of any feeder layers. The seed cells for the establishment of this line were derived from a solid tumor; the cloned cell line derived from this tumor exhibits markers characteristic of megakaryocytes and platelets. Specifically, the cells express platelet peroxidase, platelet factor 4, and platelet Ca+(+)-adenosine triphosphatase (ATPase), glycoprotein IIb- IIIa (CDw41), factor VIII antigen, and the MY7 (CD13) and MY9 (CD33) antigens. The cells do not express the erythroid markers glycophorin A and hemoglobin, the myeloid marker myeloperoxidase, nor markers specific for T and/or B cells. The established cell line produces both basic fibroblast growth factor and transforming growth factor-beta, properties demonstrated previously for the solid tumor. The clonal cell population exhibited a unique, singular karyotype, indicating cellular homogeneity. The cells display a doubling time of approximately 33 hours in either 25% horse or calf serum. Treatment of the cells with 1 X 10(-8) mol/L phorbol 12-myristate 13-acetate (PMA) leads to the induction of multi-nucleation and hyperploidy in the cells, with approximately 35% of the cells exhibiting two or more nuclei per cell, and greater than 80% of the cells enlarging in size. The establishment of this unique cell line under defined culture conditions will be beneficial for the future study of megakaryocytic properties expressed by this cell line.

scholarly journals Marrow Stromal Cell (MSC) Growth from Long Term Cryopreserved Bone Marrow.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 5227-5227 ◽  
Author(s):  
Thomas A. Lane ◽  
Davina Garls ◽  
Ellen Mackintosh ◽  
Steven C. Cramer
Keyword(s):  
Bone Marrow ◽  
Regenerative Medicine ◽  
Culture Medium ◽  
Mononuclear Cells ◽  
Primary Cultures ◽  
Metastatic Breast ◽  
Autologous Bone Marrow ◽  
Differentiation Potential ◽  
Fetal Bovine

Abstract Autologous MSC are under active investigation for a variety of potential clinical indications in regenerative medicine and gene therapy. The potential to derive autologous MSC from long-term cryopreserved bone marrow has not been fully elucidated. We report here the growth characteristics of MSC from bone marrow that was cryopreserved for more than 10 years. Methods: We identified 3 deceased patients whose cryopreserved autologous bone marrow that was originally intended for autologous transplant, was to be discarded after fulfilling all accepted criteria for discard. The pts were age 46, 47, and 60 at the time of harvest, one had high risk and 2 had metastatic breast cancer (both to bone) and all had had multiple chemotherapy regimens prior to harvest. The marrow was cryopreserved between 1994–1995 in 10% DMSO using a controlled-rate freezer and stored in the vapor phase of liquid nitrogen until June, 2006 (11–12 years). The marrow was thawed in a 37 C waterbath, diluted with an equal volume of PBS, washed × 3 in PBS, resuspended in culture medium with 20% fetal bovine serum and plated at 5 × 10E6 mononuclear cells (mnc)/cm2 in 2 – 4 T175 flasks. Culture medium was changed in 48 hours and the plates were observed during subsequent culture medium changes every 3–4 days. Results: At 2 – 4 days only scattered round cells were observed, but all samples grew scattered spindle-shaped cells were between days 4 – 12. Six of 8 primary cultures became > 70% confluent between days 11 and 21; 2 remained at < 40% confluence until replated (then achieving > 70% confluence at day 31). First passage cultures (n=12) became > 70% confluent in 4 – 10 days (median = 7) and 2nd passage cultures were > 70% confluent in 7 – 10 days (median = 9). Second or 3rd passage cultures yielded 2.1 – 4.2 × 10E6 MSC/T175 (n= 14; median 3.15) and were 89 – 96 % viable. Flow cytometry (n = 3) confirmed that 3rd passage cells stained positive for CD105 and CD73 and negative for CD14 and CD45. Additional characterization of the differentiation potential of these presumptive MSC are in progress. Conclusion: These studies indicate that cells typical for MSC can be readily grown from marrow that has been cryopreserved for > 10 years and suggest that even long-term stored bone marrow may serve as a source of cells for regenerative medicine.

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