scholarly journals Short Term Results of Fibrin Gel Obtained from Cord Blood Units: A Preliminary in Vitro Study

2019 ◽  
Vol 6 (3) ◽  
pp. 66 ◽  
Author(s):  
Panagiotis Mallis ◽  
Ioanna Gontika ◽  
Zetta Dimou ◽  
Effrosyni Panagouli ◽  
Jerome Zoidakis ◽  
...  
Keyword(s):  
Regenerative Medicine ◽  
Cord Blood ◽  
Production Process ◽  
Blood Cells ◽  
Proteomic Analysis ◽  
Platelet Rich Plasma ◽  
Multiple Reaction Monitoring ◽  
White Blood Cells ◽  
Fibrin Gel ◽  
Low Volume

Background: Recent findings have shown that the fibrin gel derived from cord blood units (CBUs) play a significant role in wound healing and tissue regeneration. The aim of this study was to standardize the fibrin gel production process in order to allow for its regular use. Methods: CBUs (n = 200) were assigned to 4 groups according to their initial volume. Then, a two-stage centrifugation protocol was applied in order to obtain platelet rich plasma (PRP). The concentration of platelets (PLTs), white blood cells (WBCs) and red blood cells (RBCs) were determined prior to and after the production process. In addition, targeted proteomic analysis using multiple reaction monitoring was performed. Finally, an appropriate volume of calcium gluconate was used in PRP for the production of fibrin gel. Results: The results of this study showed that high volume CBUs were characterized by greater recovery rates, concentration and number of PLTs compared to the low volume CBUs. Proteomic analysis revealed the presence of key proteins for regenerative medicine. Fibrin gel was successfully produced from CBUs of all groups. Conclusion: In this study, low volume CBUs could be an alternative source for the production of fibrin gel, which can be used in multiple regenerative medicine approaches.

The Effect of Red Blood Cells on the ADP-induced Aggregation of Human Platelets in Flow Through Tubes

1990 ◽  
Vol 63 (01) ◽  
pp. 112-121 ◽  
Author(s):  
David N Bell ◽  
Samira Spain ◽  
Harry L Goldsmith
Keyword(s):  
Platelet Aggregation ◽  
Shear Rate ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Platelet Rich Plasma ◽  
Mean Transit Time ◽  
White Blood Cells ◽  
Aggregate Size ◽  
Human Platelets ◽  
Induced Aggregation

SummaryThe effect of red blood cells, rbc, and shear rate on the ADPinduced aggregation of platelets in whole blood, WB, flowing through polyethylene tubing was studied using a previously described technique (1). Effluent WB was collected into 0.5% glutaraldehyde and the red blood cells removed by centrifugation through Percoll. At 23°C the rate of single platelet aggregtion was upt to 9× greater in WB than previously found in platelet-rich plasma (2) at mean tube shear rates Ḡ = 41.9,335, and 1,920 s−1, and at both 0.2 and 1.0 µM ADP. At 0.2 pM ADP, the rate of aggregation was greatest at Ḡ = 41.9 s−1 over the first 1.7 s mean transit time through the flow tube, t, but decreased steadily with time. At Ḡ ≥335 s−1 the rate of aggregation increased between t = 1.7 and 8.6 s; however, aggregate size decreased with increasing shear rate. At 1.0 µM ADP, the initial rate of single platelet aggregation was still highest at Ḡ = 41.9 s1 where large aggregates up to several millimeters in diameter containing rbc formed by t = 43 s. At this ADP concentration, aggregate size was still limited at Ḡ ≥335 s−1 but the rate of single platelet aggregation was markedly greater than at 0.2 pM ADP. By t = 43 s, no single platelets remained and rbc were not incorporated into aggregates. Although aggregate size increased slowly, large aggregates eventually formed. White blood cells were not significantly incorporated into aggregates at any shear rate or ADP concentration. Since the present technique did not induce platelet thromboxane A2 formation or cause cell lysis, these experiments provide evidence for a purely mechanical effect of rbc in augmenting platelet aggregation in WB.

scholarly journals T lymphocyte subsets of the umbilical cord blood of dogs

2009 ◽  
Vol 61 (4) ◽  
pp. 791-796 ◽  
Author(s):  
M.L.B. Cápua ◽  
A.E. Santana ◽  
A.P.M. Nakage ◽  
A.V. Godoy ◽  
A. Kataoka
Keyword(s):  
Cord Blood ◽  
Umbilical Cord ◽  
T Lymphocyte ◽  
Umbilical Cord Blood ◽  
Blood Cells ◽  
Hematological Parameters ◽  
White Blood Cells ◽  
Hemoglobin Concentration ◽  
Cd4 Cells ◽  
T Lymphocyte Subsets

The hematological parameters red blood cells (RBC) and total white blood cells (WBC) counts, hematocrit, hemoglobin concentration, and RBC indexes (median corpuscular volume and median corpuscular hemoglobin concentration) were determined and T CD5+ lymphocytes and CD4+ and CD8+ subpopulations of the umbilical cord blood (UCB) of dogs were quantified by the cytofluorimetric technique. Nine adult Beagles, from two do five-year old, were used as control. The umbilical cord blood (UCB) was collected from 20 neonate dogs. The method for the UCB collection was adequate to obtain sufficient quantity of blood for the accomplishment of the hematological analyses and lymphocyte quantification. Cytoscopic preparations of the UCB suggested high erythropoietic activity. There was no difference for the global leukocyte and lymphocyte counts between the groups. UCB T lymphocyte counts were lower than those obtained for adult dogs. The proportion of CD4:CD8 showed a great dominance of T CD4+ cells over T CD8+ lymphocytes in UCB.

scholarly journals Detection of major bcr-abl gene expression at a very low level in blood cells of some healthy individuals

Blood ◽  
1995 ◽  
Vol 86 (8) ◽  
pp. 3118-3122 ◽  
Author(s):  
C Biernaux ◽  
M Loos ◽  
A Sels ◽  
G Huez ◽  
P Stryckmans
Keyword(s):  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Blood Cells ◽  
Fusion Gene ◽  
Intensive Therapy ◽  
White Blood Cells ◽  
Healthy Adults ◽  
Rt Pcr ◽  
Total Rna

Abstract The major bcr-abl fusion gene is presently seen as the hallmark of chronic myeloid leukemia (CML) and presumably as the cause of its development. Accordingly, long-term disappearance of bcr-abl after intensive therapy is considered to be a probable cure of CML. The nested reverse transcriptase-polymerase chain reaction (RT-PCR) provides a powerful tool for minimal residual CML detection. The RT-PCR was optimized by (1) increasing the amount of total RNA involved in the reverse transcription reaction to correspond to total RNA extracted from 10(8) cells, (2) using a specific abl primer in this reverse reaction, and (3) reamplifying 10% of the RT-PCR product in nested amplification. This optimized RT-PCR permitted us to detect up to 1 copy of RNA bcr-abl synthesised in vitro, mixed with yeast RNA in an equivalent quantity to 10(8) white blood cells (WBCs). Using this highly sensitive RT-PCR during the follow-up of CML patients, a signal was unexpectedly found in healthy controls. Therefore, a systematic study of the possible expression of bcr-abl RNA in the WBCs of healthy adults and children and in umbilical cord blood was undertaken. It showed the presence of bcr-abl transcript in the blood of 22 of 73 healthy adults and in the blood of 1 of 22 children but not in 22 samples of umbilical cord blood.

scholarly journals Comprehensive proteomic analysis of white blood cells from chikungunya fever patients of different severities

2014 ◽  
Vol 12 (1) ◽  
pp. 96 ◽  
Author(s):  
Nitwara Wikan ◽  
Sarawut Khongwichit ◽  
Weerawat Phuklia ◽  
Sukathida Ubol ◽  
Tipparat Thonsakulprasert ◽  
...  
Keyword(s):  
Blood Cells ◽  
Proteomic Analysis ◽  
White Blood Cells ◽  
Chikungunya Fever

scholarly journals Detection of major bcr-abl gene expression at a very low level in blood cells of some healthy individuals

Blood ◽  
1995 ◽  
Vol 86 (8) ◽  
pp. 3118-3122 ◽  
Author(s):  
C Biernaux ◽  
M Loos ◽  
A Sels ◽  
G Huez ◽  
P Stryckmans
Keyword(s):  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Blood Cells ◽  
Fusion Gene ◽  
Intensive Therapy ◽  
White Blood Cells ◽  
Healthy Adults ◽  
Rt Pcr ◽  
Total Rna

The major bcr-abl fusion gene is presently seen as the hallmark of chronic myeloid leukemia (CML) and presumably as the cause of its development. Accordingly, long-term disappearance of bcr-abl after intensive therapy is considered to be a probable cure of CML. The nested reverse transcriptase-polymerase chain reaction (RT-PCR) provides a powerful tool for minimal residual CML detection. The RT-PCR was optimized by (1) increasing the amount of total RNA involved in the reverse transcription reaction to correspond to total RNA extracted from 10(8) cells, (2) using a specific abl primer in this reverse reaction, and (3) reamplifying 10% of the RT-PCR product in nested amplification. This optimized RT-PCR permitted us to detect up to 1 copy of RNA bcr-abl synthesised in vitro, mixed with yeast RNA in an equivalent quantity to 10(8) white blood cells (WBCs). Using this highly sensitive RT-PCR during the follow-up of CML patients, a signal was unexpectedly found in healthy controls. Therefore, a systematic study of the possible expression of bcr-abl RNA in the WBCs of healthy adults and children and in umbilical cord blood was undertaken. It showed the presence of bcr-abl transcript in the blood of 22 of 73 healthy adults and in the blood of 1 of 22 children but not in 22 samples of umbilical cord blood.

scholarly journals Estimation of Eosinophil Cells in Cord Blood with References Based on Blood in Adults via Bayesian Measurement Error Modeling

2019 ◽  
Author(s):  
Yu Jiang ◽  
Hongmei Zhang ◽  
Shan V Andrews ◽  
Hasan Arshad ◽  
Susan Ewart ◽  
...  
Keyword(s):  
Measurement Error ◽  
Cord Blood ◽  
Blood Cells ◽  
White Blood Cells ◽  
Error Modeling ◽  
Supplementary Information ◽  
Estimation Accuracy ◽  
Bioconductor Package ◽  
High Quality ◽  
Eosinophil Cell

Abstract Motivation Eosinophils are phagocytic white blood cells with a variety of roles in the immune system. In situations where actual counts are not available, high quality approximations of their cell proportions using indirect markers are critical. Results We develop a Bayesian measurement error model to estimate proportions of eosinophils in cord blood, using the cord blood DNA methylation profiles, based on markers of eosinophil cell heterogeneity in blood of adults. The proposed method can be directly extended to other cells across different reference panels. We demonstrate the method’s estimation accuracy using B cells and show that the findings support the proposed approach. The method has been incorporated into the estimateCellCounts function in the minfi package to estimate eosinophil cells proportions in cord blood. Availability estimateCellCounts function is implemented and available in Bioconductor package minfi. Supplementary information Supplementary data are available at Bioinformatics online.

scholarly journals Role of White Blood Cells in Blood- and Bone Marrow-Based Autologous Therapies

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
William King ◽  
Krista Toler ◽  
Jennifer Woodell-May
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Growth Factors ◽  
Blood Cells ◽  
Platelet Rich Plasma ◽  
White Blood Cells ◽  
Complex Mixture ◽  
Negative Results

There has been significant debate over the role of white blood cells (WBCs) in autologous therapies, with several groups suggesting that WBCs are purely inflammatory. Misconceptions in the practice of biologic orthopedics result in the simplified principle that platelets deliver growth factors, WBCs cause inflammation, and the singular value of bone marrow is the stem cells. The aim of this review is to address these common misconceptions which will enable better development of future orthopedic medical devices. WBC behavior is adaptive in nature and, depending on their environment, WBCs can hinder or induce healing. Successful tissue repair occurs when platelets arrive at a wound site, degranulate, and release growth factors and cytokines which, in turn, recruit WBCs to the damaged tissue. Therefore, a key role of even pure platelet-rich plasma is to recruit WBCs to a wound. Bone marrow contains a complex mixture of vascular cells, white blood cells present at much greater concentrations than in blood, and a small number of progenitor cells and stem cells. The negative results observed for WBC-containing autologous therapies in vitro have not translated to human clinical studies. With an enhanced understanding of the complex WBC biology, the next generation of biologics will be more specific, likely resulting in improved effectiveness.

Differential Stability Of Prostacyclin (PGI2) In Whole Blood And Plasma

1981 ◽  
Author(s):  
S Krishnamurthi ◽  
J Westwick ◽  
V V Kakkar
Keyword(s):  
Whole Blood ◽  
Blood Cells ◽  
Time Course ◽  
Platelet Rich Plasma ◽  
White Blood Cells ◽  
Platelet Inhibition ◽  
Solvent Systems ◽  
Differential Stability ◽  
Significant Uptake ◽  
The Stability

The stability of PGI2 in human whole blood (WB) , platelet- rich plasma (PRP) and platelet-poor plasma (PPP) was studied. Following incubation of PGI2(60nM) in the three media, it was found that the half life (t 1/2) of PGI2 (as measured) by the rate of loss of PGI2 equivalents causing platelet inhibition in the supernatant of the incubates) was longer in PPP ᶓ PRP (t 1/2 49±4 ᶓ 42.5±5 min respectively, n=10). On investigation this was found to be largely due to the pH difference observed between PPPᶓPRP (both pH 7.8±1) and WB (pH 7.4±1). Addition of NaHCO3 to raise the pH of WB to 7.8 prolonged the stability of PGI2 with a th of 35±4 min and addition of HC1 to lower the pH of PPP to 7.4 shortened the th to 18.5±4 min. However, incubation of PGI2 in either Hanks buffer or washed red blood cells (WRBC) at pH 7.8 did not increase PG2 stability. Since addition of a mixed population of white blood cells (7xl06cells/ml in PBS pH 7.8) to PPP (pH7.8) did not alter the rate of loss of PGI2 activity and there was found to be no significant uptake (<15%) of 3H PGI2 in WRBC, the possibility of PGI2 conversion to a more stable and platelet-active metabolite such as 6-oxo-PGE2 in plasma was studied by extraction and TLC of the PPP and WB incubates. 3H PGI 2 was found to be converted to % 6-oxo-PGF2 in both WB (pH7.4) and PPP (pH 7.8) with no other detectable metabolites in three different solvent systems. Treatment of the incubates prior to extraction and TLC with NaBH4 (which by reducing free keto groups can distinguish between PGI2 and 6-oxo-PGF1α) showed that 10-20% of the added 3H PGI2 in PPP (pH 7.8) was unchanged even after 120 mins incubation while virtually all the added 3H PGI2 in WB (pH7.4) was converted to 3H 6-oxo-PGFlaby 50 min with a time course (% PGI2 60nM; 1/2 WB-14 min, PPP-35min) similar to the loss of PGI2 activity in WB and PPP on bioassay. We conclude that the prolonged platelet inhibitory activity following incubation of PGI2 in plasma compared to that in whole blood is due to unchanged PGI2 and not the formation of a 6-oxo-PGE1, like substance as suggested by Borda and Gimeno in Prostaglandins 19 pp 899 (1979).

scholarly journals Leucocyte-Rich Platelet-Rich Plasma Enhances Fibroblast and Extracellular Matrix Activity: Implications in Wound Healing

2020 ◽  
Vol 21 (18) ◽  
pp. 6519
Author(s):  
Jeannie Devereaux ◽  
Narges Dargahi ◽  
Sarah Fraser ◽  
Kulmira Nurgali ◽  
Dimitrios Kiatos ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cell Proliferation ◽  
Wound Healing ◽  
Blood Cells ◽  
Platelet Rich Plasma ◽  
Autologous Blood ◽  
Human Fibroblasts ◽  
White Blood Cells ◽  
High Concentration ◽  
Gene Expressions

Background: Platelet-rich plasma (PRP) is an autologous blood product that contains a high concentration of platelets and leucocytes, which are fundamental fibroblast proliferation agents. Literature has emerged that offers contradictory findings about leucocytes within PRP. Herein, we elucidated the effects of highly concentrated leucocytes and platelets on human fibroblasts. Methods: Leucocyte-rich, PRP (LR-PRP) and leucocyte-poor, platelet-poor plasma (LP-PPP) were compared to identify their effects on human fibroblasts, including cell proliferation, wound healing and extracellular matrix and adhesion molecule gene expressions. Results: The LR-PRP exhibited 1422.00 ± 317.21 × 103 platelets/µL and 16.36 ± 2.08 × 103 white blood cells/µL whilst the LP-PPP demonstrated lower concentrations of 55.33 ± 10.13 × 103 platelets/µL and 0.8 ± 0.02 × 103 white blood cells/µL. LR-PRP enhanced fibroblast cell proliferation and cell migration, and demonstrated either upregulation or down-regulation gene expression profile of the extracellular matrix and adhesion molecules. Conclusion: LR-PRP has a continuous stimulatory anabolic and ergogenic effect on human fibroblast cells.

Performance evaluation of Sysmex XN hematology analyzer in umbilical cord blood: a comparison study with Sysmex XE-2100

2014 ◽  
Vol 52 (12) ◽  
Author(s):  
Hanah Kim ◽  
Mina Hur ◽  
Sang-Gyeu Choi ◽  
Hee-Won Moon ◽  
Yeo-Min Yun ◽  
...  
Keyword(s):  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Blood Cells ◽  
Complete Blood Count ◽  
White Blood Cells ◽  
Modular System ◽  
Hematopoietic Stem ◽  
Nucleated Red Blood Cells ◽  
Hematology Analyzer

AbstractThe Sysmex XN (XN) modular system (Sysmex, Kobe, Japan) is a new automated hematology analyzer equipped with different principles from its previous version, Sysmex XE-2100. We compared the performances of Sysmex XN and XE-2100 in umbilical cord blood (CB) specimens.In 160 CB specimens, complete blood count (CBC) parameters and white blood cells (WBC) differentials were compared between the two analyzers. Their flagging performances for blasts, abnormal/atypical lymphocytes, immature granulocytes and/or left-shift (IG), and nucleated red blood cells (NRBC) counts were compared with manual counts. For the blast flagging, Q values by Sysmex XN were further compared with manual slide review.Sysmex XN and XE-2100 showed high or very high correlations for most CBC parameters but variable correlations for WBC differentials. Compared with XE-2100, XN showed significantly different flagging performances for blasts, abnormal/atypical lymphocytes, and IG. The flagging efficiency for blasts was significantly better on Sysmex XN than on XE-2100 (85.0% vs. 38.8%): Sysmex XN showed a remarkably increased specificity of blast flag, compromising its sensitivity of blast flag. Among the 24 specimens with blasts (range, 0.5%–1.5%), only one (4.2%) showed a positive Q value.This study highlighted the remarkable differences of flagging performances between Sysmex XN and XE-2100 in CB specimens. The Sysmex XN modular system seems to be a suitable and practical option for the CB specimens used for hematopoietic stem cell transplantation as well as for the specimens from neonates.

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