Amnion and Umbilical Cord–Derived Products in Sports Medicine: From Basic Science to Clinical Application

2021 ◽  
pp. 036354652110104
Author(s):  
Lisa A. Fortier ◽  
Marta Cercone ◽  
Laura E. Keller ◽  
Michelle L. Delco ◽  
Lilia Becktell ◽  
...  
Keyword(s):  
Health Care ◽  
Stem Cells ◽  
Flow Cytometry ◽  
Growth Factors ◽  
Cell Viability ◽  
Cord Blood ◽  
Health Care Professionals ◽  
Blood Product ◽  
Biological Tissues ◽  
Cell Type

Background: Birth tissue products from amnion, chorion, umbilical cord, amniotic fluid, or cord blood are frequently marketed as viable sources of stem cells and growth factors. It can be difficult for health care professionals to differentiate implied from explicit conclusions in reported product analyses. Purpose: To provide an educational platform for health care professionals to interpret data presented in the promotion of birth tissue products. Study Design: Descriptive laboratory study and expert opinion; Level of evidence, 5. Methods: A cord blood product was analyzed by 3 methods for cell viability, 2 methods for assessment of cell morphology and cell type, multicolor flow cytometry to identify stem cells, and enzyme-linked immunosorbent assay (ELISA) plus Western blot for analysis of interleukin 1 receptor antagonist protein (IL-1ra). These data were compared with analyses reported by the manufacturer. Results: Cell viability in the cord blood product was less than reported by the manufacturer, the cells were primarily leukocytes, no stem cells were present, and the concentration of IL-1ra was falsely increased due to nonspecific antibody binding in the sample. Conclusion: To assess birth tissue products, health care professionals should consider the following: (1) Understanding fluorescent dyes is important for assessing cell viability data—green does not always mean alive. (2) The report of “cells” in the product does not necessarily mean “stem cells”; microscopic images of at least ×20 or a hemogram should be evaluated to determine cell type (leukocyte, red blood cells, etc). (3) There is no single cluster of differentiation (CD) marker on flow cytometry to identify stem cells. (4) Biological tissues are complex substances, and inaccurately increased measurements of growth factors could be present in ELISA results because most ELISAs are not designed or validated for use in biologics. Furthermore, the reported measurement of growth factors should be considered relative to concentrations in native biological tissues and plasma. Clinical Relevance: Health care professionals should be able to interpret cell viability, cell morphology, stem cell analysis using CD markers, and growth factor analysis when considering use of a birth tissue product in patients.

scholarly journals Contemporary Transfusion Science and Challenges

2019 ◽  
Vol 30 (2) ◽  
pp. 139-150
Author(s):  
Heather M. Passerini
Keyword(s):  
Health Care ◽  
Health Care Professionals ◽  
Blood Product ◽  
Damage Control ◽  
Coagulation Factor ◽  
Trauma Patients ◽  
Transfusion Therapy ◽  
Use Of Resources ◽  
Study Results ◽  
The Impact

Health care professionals must understand the impact of blood product transfusions and transfusion therapy procedures to ensure high-quality patient care, positive outcomes, and wise use of resources in blood management programs. Understanding transfusions of blood and blood products is also important because of the number of treatments performed, which affects individual patients and health care system resources. This article reviews research findings to acquaint health care professionals with the most successful protocols for blood, blood product, and coagulation factor transfusions. Damage control resuscitation in bleeding trauma patients, protocols for patients without trauma who are undergoing surgical procedures that place them at risk for excessive bleeding, and protocols for patients with sepsis are addressed. Emerging research continues to help guide mass transfusion treatments (restrictive vs liberal, balanced, and goal-directed treatment). Although available study results provide some guidance, questions remain. Additional research by health care professionals is needed.

Percoll Gradient Separation of Cord Blood Mononuclear Cells Reveals the Presence of a Novel Population of CXCR4+ Oct-4+ Small Embryonic-Like Stem Cells.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1069-1069 ◽  
Author(s):  
Magda Kucia ◽  
Krzysztof Oldak ◽  
Mariusz Z. Ratajczak ◽  
Janina Ratajczak ◽  
Zygmunt Pojda
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Cord Blood ◽  
Intestinal Epithelium ◽  
Skeletal Muscles ◽  
Mononuclear Cells ◽  
Neural Tissue ◽  
Open Chromatin ◽  
Percoll Gradient ◽  
Cell Type

Abstract Cord blood (CB) mononuclear cells (MNC) were demonstrated to contribute to organ/tissue regeneration, however, the identity of the specific cell type(s) involved remains unknown. Recently, it had been identified in adult bone marrow (BM) a mobile, SDF-1-, HGF/SF- and LIF-responsive population of CXCR4+ non-hematopoietic MNC that expresses markers (RQ-PCR, immunhistochemistry) of early pluripotent/tissue committed stem cells (TCSC) for skeletal muscles, heart, neural tissue, liver, pancreas, epidermis and intestinal epithelium (Leukemia2004:18;29–40). We hypothesized that a similar population of these rare cells may also be present in CB, however, their final yield may depend on the method of MNC preparation. We hypothesized that since these cells are very small (~3–5 μm), they may cross a Ficoll-Paque gradient or sedimentate more rapidly than other CB MNC and as result of this are lost during routine CB preparations. Thus, taking in consideration their small size, in order to evaluate if these cells are present in CB, we isolated small CB MNC by employing Percoll gradient (1.078–1.095). We found that this allows us to isolate from CB a population of small cells (2.5% of the total number of MNC) that is enriched in a population of TCSC/PSC(~0.002% of MNC) that we have originally identified in BM. Accordingly, these CB-derived TCSC/PSC CXCR4+ cells are very small (~3μm), posses large nuclei that contain embryonic stem cell type open chromatin (euchromatin), and express several markers for skeletal muscles, heart, neural tissue, liver, pancreas, epidermis and intestinal epithelium as well as pluripotent stem cell (PSC) transcription factors such as Oct-4, Nanog and Rex-1. In vitro cultures of CB-derived small TCSC/PSC are able to grow neurospheres that gave rise to neuronal (β-III tubulin+, nestin+) and macroglia (O4+, MBP+, GFAP+) lineages and cardiomyocytes (β-myosin heavy chain+, α-sarcomeric actin+). Based on this we conclude that CB contains embryonic-like stem cells which may be lost during routine procedures to isolate MNC. Thus, Percoll gradient centrifugation allows for optimal isolation of these small CXCR4+ PSC/TCSC and we postulate that the CB tissue/organ regenerating potential may be much higher than initially postulated and we are currently testing this hypothesis in vivo in animal models.

Substance P and calcitonin gene-related neuropeptides as novel growth factors forex vivoexpansion of cord blood CD34+hematopoietic stem cells

2009 ◽  
Vol 28 (1) ◽  
pp. 66-73 ◽  
Author(s):  
Somayeh Shahrokhi ◽  
Massoumeh Ebtekar ◽  
Kamran Alimoghaddam ◽  
Ali Akbar Pourfathollah ◽  
Maryam Kheirandish ◽  
...  
Keyword(s):  
Stem Cells ◽  
Growth Factors ◽  
Substance P ◽  
Hematopoietic Stem Cells ◽  
Cord Blood ◽  
Hematopoietic Stem ◽  
Calcitonin Gene ◽  
Cord Blood Cd34

Improved cord blood thawing procedure enhances the reproducibility and correlation between flow cytometry CD34 + cell viability and clonogenicity assays

Cytotherapy ◽  
2018 ◽  
Vol 20 (6) ◽  
pp. 891-894 ◽  
Author(s):  
Cristian Camilo Galindo ◽  
Diana María Vanegas Lozano ◽  
Bernardo Camacho Rodríguez ◽  
Ana-María Perdomo-Arciniegas
Keyword(s):  
Flow Cytometry ◽  
Cell Viability ◽  
Cord Blood ◽  
Thawing Procedure

scholarly journals Growth factors-based beneficial effects of platelet lysate on umbilical cord-derived stem cells and their synergistic use in osteoarthritis treatment

2020 ◽  
Vol 11 (10) ◽  
Author(s):  
Li Yan ◽  
Li Zhou ◽  
Bo Yan ◽  
Lei Zhang ◽  
Wenxi Du ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Flow Cytometry ◽  
Growth Factors ◽  
Umbilical Cord ◽  
Control Flow ◽  
Platelet Lysate ◽  
Specific Staining ◽  
Beneficial Effects

Abstract Poor viability of mesenchymal stem cells (MSCs) at the transplanted site often hinders the efficacy of MSCs-based therapy. Platelet lysate (PL) contains rich amounts of growth factors, which benefits cell growth. This study aimed to explore how human PL benefits umbilical cord-derived MSCs (huc-MSCs), and whether they have synergistic potential in osteoarthritis (OA) treatment. As quality control, flow cytometry and specific staining were performed to identify huc-MSCs, and ELISA was used to quantify growth factors in PL. CCK-8 and flow cytometry assays were performed to evaluate the effects of PL on the cell viability and cell cycle progression of huc-MSCs. Wound healing and transwell assays were conducted to assess the migration of huc-MSCs. RNA sequencing, real time PCR, and Western blot assays were conducted to explore the growth factors-based mechanism of PL. The in vitro results showed that PL significantly promoted the proliferation, cell cycle, and migration of huc-MSCs by upregulating relevant genes/proteins and activating beclin1-dependent autophagy via the AMPK/mTOR signaling pathway. The main growth factors (PDGF-AA, IGF-1, TGF-β, EGF, and FGF) contributed to the effects of PL in varying degrees. The in vivo data showed that combined PL and huc-MSCs exerted significant synergistic effect against OA. The overall study determined the beneficial effects and mechanism of PL on huc-MSCs and indicated PL as an adjuvant for huc-MSCs in treating OA. This is the first report on the growth factors-based mechanism of PL on huc-MSCs and their synergistic application. It provides novel knowledge of PLʹs roles and offers a promising strategy for stem cell-based OA therapy by combining PL and huc-MSCs.

Effect of rat bone marrow derived-mesenchymal stem cells on granulocyte differentiation of mononuclear cells as preclinical agent in cell-based therapy

2021 ◽  
Vol 21 ◽  
Author(s):  
Ezzatollah Fathi ◽  
Sheyda Azarbad ◽  
Raheleh Farahzadi ◽  
Sara Javanmardi
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Growth Factors ◽  
Mononuclear Cells ◽  
Control Group ◽  
Flow Cytometry Analysis ◽  
Cell Lineages ◽  
Experimental Group

Background: Bone marrow mononuclear cells (BM-MNCs), as a collection of hematopoietic and mesenchymal stem cells (MSCs), are capable of producing all blood cell lineages. The use of cytokines, growth factors, or cells capable of secreting these factors will help in stimulating the proliferation and differentiation of these cells into mature cell lines. On the other hand, MSCs are multipotent stromal cells that can be differentiated into various cell lineages. Moreover, these cells can control the process of hematopoiesis by secreting cytokines and growth factors. The present study aimed to investigate the effect of BM-derived MSCs on the differentiation of MNCs based on the assessment of cell surface markers by flow cytometry analysis. Methods: For this purpose, the MNCs were purified from rat BM using density gradient centrifugation. After that, they were cultured, expanded, and characterized. Next, BM-derivedMSCs were co-cultured with MNCs and then were either cultured with MNCs alone (control group) or co-cultured MNCs with BM derived-MSCs (experimental group). Finally, they were collected on day 7 and subjected to flow cytometry analysis for granulocyte markers and ERK protein’s investigation. Results: It was found that the expression levels of CD34, CD16, CD11b, and CD18 granulocyte markers, as well as protein expression of ERK, have significantly increased in the experimental group compared to the control group. Conclusion: Therefore, it can be concluded that MSCs could affect the granulocyte differentiation of MNCs via ERK protein expression, which is a key component of the ERK signaling pathway.

Increased Engraftment after Double Cord Blood Transplantation in NOD/SCID Mice.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1199-1199
Author(s):  
Alma J. Nauta ◽  
Alwine B. Kruisselbrink ◽  
Roelof Willemze ◽  
Willem E. Fibbe
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
Cord Blood ◽  
Cord Blood Transplantation ◽  
Human Platelets ◽  
Hematopoietic Stem ◽  
Cell Dose ◽  
Blood Transplantation ◽  
Cd34 Cells

Abstract Umbilical cord blood (UCB) is considered as an attractive alternative source of hematopoietic stem cells for allogeneic stem cell transplantations in patients who lack HLA-matched donors. However, the low cell dose adversely affects the speed of hematopoietic recovery and therefore limits the application of UCB transplantation in adults. Although ex-vivo expansion of cord blood cells has been explored as a strategy to increase the cell dose, compromised engraftment potential of expanded cells has been demonstrated. Another approach to overcome cell dose limitations is transplantation of multiple, unrelated UCB units. To investigate the effect of multiple cord transplantation on engraftment, NOD/SCID mice were transplanted with human hematopoietic progenitor cells (CD34+) derived from two UCB units with HLA disparity. During the first six weeks after transplantation the number of human platelets in peripheral blood was quantified by flow cytometry. Six weeks after transplantation, the mice were sacrificed and the percentage and donor origin of human CD45+ cells in blood, and in bone marrow was determined by flow cytometry. Transplantation of CD34+ cells derived from two UCB donors resulted in significantly higher number of human platelets in peripheral blood than transplantation of CD34+ cells from either donor alone, ranging from 3.92x106/ml to 10.29x106/ml (mean 6.4x106 ± 2.55x106/ml) and 0.11x106/ml to 3.12.106/ml (mean 1.42x106 ± 1.17x106/ml), respectively. Furthermore, the overall human cell engraftment level in bone marrow after double cord blood transplantation ranged from 7.01% to 64.34% (mean 29.6 ± 21.5%) a nearly 7-fold increase compared to single cord blood transplantation ranging from 0.27% to 13.5% (mean 4.6 ± 3.8%) Although consistently higher engraftment levels were reached after double cord blood transplantation, two different patterns were observed: in 2 out of 4 experiments cells from one donor predominated the engraftment (ratio 3:1), while in two other experiments the two units contributed equally to BM engraftment. The mechanism underlying these effects are <S>is</S> not yet clear. It is not very likely that the single donor predominance results from an unequal amount of hematopoietic stem cells in the cord blood units because each cord blood showed comparable levels of engraftment as a single unit. Alternatively, the unequal engraftment may result from an immunological competition or a graft versus graft stimulatory effect between the cords during the engraftment process and further studies are required to determine if the contribution of both units is dependent on the degree of HLA matching between the two cords. Taken together, these results demonstrate that double cord blood transplantation may represent a means of achieving increased engraftment, making multiple cord blood transplantation a promising strategy to improve the outcome of UCB transplantation. Studies are underway to unravel the mechanisms underlying the enhanced engraftment.

Sign in / Sign up

Export Citation Format

Share Document