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Article: Cell-Free Amniotic Fluid and Regenerative Medicine: Current Applications and Future Opportunities.

Bowen, Charles M / Ditmars, Frederick S / Gupta, Ashim / Reems, Jo-Anna / Fagg, William Samuel

2022 Volume 10, Issue 11

Abstract: Amniotic fluid (AF) provides critical biological and physical support for the developing fetus. While AF is an excellent source of progenitor cells with regenerative properties, recent investigations indicate that cell-free AF (cfAF), which consists of ... ...

Abstract	Amniotic fluid (AF) provides critical biological and physical support for the developing fetus. While AF is an excellent source of progenitor cells with regenerative properties, recent investigations indicate that cell-free AF (cfAF), which consists of its soluble components and extracellular vesicles, can also stimulate regenerative and reparative activities. This review summarizes published fundamental, translational, and clinical investigations into the biological activity and potential use of cfAF as a therapeutic agent. Recurring themes emerge from these studies, which indicate that cfAF can confer immunomodulatory, anti-inflammatory, and pro-growth characteristics to the target cells/tissue with which they come into contact. Another common observation is that cfAF seems to promote a return of cells/tissue to a homeostatic resting state when applied to a model of cell stress or disease. The precise mechanisms through which these effects are mediated have not been entirely defined, but it is clear that cfAF can safely and effectively treat cutaneous wounds and perhaps orthopedic degenerative conditions. Additional applications are currently being investigated, but require further study to dissect the fundamental mechanisms through which its regenerative effects are mediated. By doing so, rational design can be used to fully unlock its potential in the biotechnology lab and in the clinic.
Language	English
Publishing date	2022-11-17
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2720867-9
ISSN	2227-9059
ISSN	2227-9059
DOI	10.3390/biomedicines10112960
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Article ; Online: A journey to produce platelets in vitro.

Reems, Jo-Anna

Transfusion

2011 Volume 51 Suppl 4, Page(s) 169S–176S

Abstract: Allogeneic platelet transfusions protect patients from bleeding episodes and also make aggressive medical procedures such as those involving marrow transplants requiring chemotherapy and/or radiotherapy possible. These patients are dependent upon an ... ...

Abstract	Allogeneic platelet transfusions protect patients from bleeding episodes and also make aggressive medical procedures such as those involving marrow transplants requiring chemotherapy and/or radiotherapy possible. These patients are dependent upon an unfailing supply of platelets that can sometimes be in short supply due to high demands coupled with an extremely short expiration date for platelet products of only 5 days. One approach that is under investigation to overcome platelet shortages is to harness the extraordinary capabilities of stem cells to proliferate and differentiate into various cell types and to use this ability to specifically produce clinical scale quantities of functional platelets in bioreactors. To accomplish such an enormous and complex task requires an appreciation of the regulatory mechanisms that occur during the development of megakaryocytes (MKs) and the subsequent biogenesis of functional platelets from mature MKs. This means understanding the complex network of intracellular and extracellular regulatory mechanisms that act at each phase of a developmental process that ushers stem cells along the MK lineage to produce billions of platelets per day in a healthy individual.
MeSH term(s)	Antigens, Human Platelet/biosynthesis ; Bioreactors ; Blood Platelets/cytology ; Cell Culture Techniques ; Cell Separation/methods ; Cells, Cultured/cytology ; Cells, Cultured/drug effects ; Culture Media/pharmacology ; Cytokines/pharmacology ; Hematopoietic Stem Cells/cytology ; Hematopoietic Stem Cells/drug effects ; Humans ; Megakaryocytes/cytology ; Megakaryocytes/drug effects ; Platelet Transfusion/methods ; Thrombopoiesis/drug effects ; Thrombopoietin/pharmacology
Chemical Substances	Antigens, Human Platelet ; Culture Media ; Cytokines ; Thrombopoietin (9014-42-0)
Language	English
Publishing date	2011-11
Publishing country	United States
Document type	Journal Article
ZDB-ID	208417-x
ISSN	1537-2995 ; 0041-1132
ISSN (online)	1537-2995
ISSN	0041-1132
DOI	10.1111/j.1537-2995.2011.03380.x
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Article ; Online: Local manufacturing processes contribute to variability in human mesenchymal stromal cell expansion while growth media supplements contribute to variability in gene expression and cell function: a Biomedical Excellence for Safer Transfusion (BEST) collaborative study.

Shaz, Beth H / Schäfer, Richard / Fontaine, Magali J / Norris, Philip J / McKenna, David H / Jin, Ping / Reems, Jo-Anna / Stroncek, David / Tanashi, Minoko / Marks, Denese / Geng, Huimin / Pati, Shibani

Cytotherapy

2023

Abstract: Background aims: Culture-derived mesenchymal stromal cells (MSCs) exhibit variable characteristics when manufactured using different methods, source material and culture media. The purpose of this multicenter study was to assess the impact on MSC ... ...

Abstract	Background aims: Culture-derived mesenchymal stromal cells (MSCs) exhibit variable characteristics when manufactured using different methods, source material and culture media. The purpose of this multicenter study was to assess the impact on MSC expansion, gene expression and other characteristics when different laboratories expanded MSCs from cultures initiated with bone marrow-MSC aliquots derived from the same donor source material yet with different growth media. Methods: Eight centers expanded MSCs using four human platelet lysate (HPL) and one fetal bovine serum (FBS) products as media supplements. The expanded cells were taken through two passages then assessed for cell count, viability, doubling time, immunophenotype, cell function, immunosuppression and gene expression. Results were analyzed by growth media and by center. Results: Center methodologies varied by their local seeding density, feeding regimen, inoculation density, base media and other growth media features (antibiotics, glutamine, serum). Doubling times were more dependent on center than on media supplements. Two centers had appropriate immunophenotyping showing all MSC cultures were positive for CD105, CD73, CD90 and negative for CD34, CD45, CD14, HLA-DR. MSCs cultured in media supplemented with FBS compared with HPL featured greater T-cell inhibition potential. Gene expression analysis showed greater impact of the type of media supplement (HPL versus FBS) than the manufacturing center. Specifically, nine genes were decreased in expression and six increased when combining the four HPL-grown MSCs versus FBS (false discovery rate [FDR] <0.01), however, without significant difference between different sources of HPL (FDR <0.01). Conclusions: Local manufacturing process plays a critical role in MSC expansion while growth media may influence function and gene expression. All HPL and FBS products supported cell growth.
Language	English
Publishing date	2023-12-02
Publishing country	England
Document type	Journal Article
ZDB-ID	2039821-9
ISSN	1477-2566 ; 1465-3249
ISSN (online)	1477-2566
ISSN	1465-3249
DOI	10.1016/j.jcyt.2023.11.003
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Article: Amnion membrane allografts in a critically ill infant with Netherton syndrome-like phenotype.

Frigerio, Alice / Bleicher, Josh / Pierce, Jan / Reems, Jo-Anna / Vanderhooft, Sheryll L / Lewis, Giavonni

JAAD case reports

2019 Volume 5, Issue 5, Page(s) 395–397

Language	English
Publishing date	2019-04-20
Publishing country	United States
Document type	Case Reports
ZDB-ID	2834220-3
ISSN	2352-5126
ISSN	2352-5126
DOI	10.1016/j.jdcr.2019.02.011
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Article ; Online: Red Cell Depletion of Major ABO Incompatible Bone Marrow Hematopoietic Progenitor Cells HPC(M) with the Spectra Optia®.

Christensen, Austin / Hsieh, FenFen / Boyer, Michael / Couriel, Daniel / Reems, Jo-Anna

Annals of clinical and laboratory science

2020 Volume 50, Issue 6, Page(s) 781–789

Abstract: Objective: Major ABO incompatible hematopoietic progenitors from bone marrow (HPC(M)) donor collections that are destined for clinical transplantation are typically processed to deplete products of red blood cells (RBCs). The purpose of this study was ... ...

Abstract	Objective: Major ABO incompatible hematopoietic progenitors from bone marrow (HPC(M)) donor collections that are destined for clinical transplantation are typically processed to deplete products of red blood cells (RBCs). The purpose of this study was to compare RBC depletion when using the Spectra Optia® relative to a 2-step method involving a COBE2991 instrument to obtain a buffy coat followed by a hydroxyethyl starch (HES) density gradient (COBE+HES) of the buffy coat. Methods: Post-processing recoveries of products undergoing 4, 8, and 10 bone marrow processing (BMP) cycles (i.e. 1 cycle=1 volume of HPC(M)) with the Spectra Optia® were determined for volume, RBC content, viable total nucleated cells (vTNC), viable CD34+ cells (vCD34), viable CD3+ cells (vCD3) and colony-forming-cells (CFC). Subsequent RBC depletions with Spectra Optia® were then performed with 10 BMP cycles on additional HPC(M) collections and were compared against a retrospective analysis of historical COBE+HES post-processing data. Results: Ten BMP cycles of HPC(M) (n=6) products were identified as optimal with volume reductions of 81.3±1.6 % and RBC reductions of 97.0±0.6 % with the Spectra Optia®. This also resulted in an average of 0.28 ±0.14 mL of RBC/kg (mean±SD; n=6) with vTNC yields of 65.0±10.9%, vCD34+ yields of 98.5±12.7%, and vCD3+ yields of 90.6±10.0%. Recoveries with the COBE+HES methodology resulted in vTNC recoveries of 62.9±20.5% (mean±SD; n=30) and 0.63±0.71 mL of RBC/kg (mean±SD; n=30). Conclusions: The Spectra Optia® is a viable option for depleting HPC(M) harvests of contaminating RBC in situations of ABO incompatibility. Target cells from a MNC rich fractionation were preserved through processing while eliminating RBC contaminants.
MeSH term(s)	ABO Blood-Group System/metabolism ; Blood Component Removal/methods ; Bone Marrow/immunology ; Bone Marrow/metabolism ; Bone Marrow Transplantation/methods ; Cell Separation/methods ; Erythrocytes/immunology ; Healthy Volunteers ; Hematopoietic Stem Cell Transplantation/methods ; Hematopoietic Stem Cells/immunology ; Hematopoietic Stem Cells/metabolism ; Hematopoietic Stem Cells/pathology ; Humans ; Retrospective Studies ; Tissue Donors
Chemical Substances	ABO Blood-Group System
Language	English
Publishing date	2020-12-17
Publishing country	United States
Document type	Comparative Study ; Journal Article
ZDB-ID	193092-8
ISSN	1550-8080 ; 0091-7370 ; 0095-8905
ISSN (online)	1550-8080
ISSN	0091-7370 ; 0095-8905
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Article ; Online: Processed human amniotic fluid retains its antibacterial activity.

Mao, Yong / Pierce, Jan / Singh-Varma, Anya / Boyer, Michael / Kohn, Joachim / Reems, Jo-Anna

Journal of translational medicine

2019 Volume 17, Issue 1, Page(s) 68

Abstract: Background: Human amniotic fluid (AF) contains numerous nutrients, trophic factors and defense proteins that provide a nurturing and protective environment for fetal development. Based on reports that AF has antibacterial, anti-inflammatory and ... ...

Abstract	Background: Human amniotic fluid (AF) contains numerous nutrients, trophic factors and defense proteins that provide a nurturing and protective environment for fetal development. Based on reports that AF has antibacterial, anti-inflammatory and regenerative properties, we designed a novel method to process AF for use in clinical care. Methods: Six randomly selected lots of processed AF (pAF) were examined to determine whether they retained their antibacterial activity against a panel of wound-associated pathogens E. faecium, S. aureus, K. pneumoniae, A. baumannii, P. aeruginosa, and E. aerogenes (ESKAPE). To identify proteins in pAF that might be responsible for its antibacterial activity, three different lots of pAF were analyzed with quantitative cytokine arrays that consisted of 400 unique human proteins. One protein identified by microarrays, lactoferrin, and a second prominent antibacterial protein that was not identified by microarrays, lysozyme, were examined by depletion experiments to determine their contribution to the antibacterial activity of pAF. Results: All six lots of pAF exhibited antibacterial activity against ESKAPE microorganisms, especially against the pathogens predominately found in chronic wounds (i.e. S. aureus and P. aeruginosa). Thirty-one of the peptides on the microarray were annotated as having antibacterial activity and 26 of these were detected in pAF. Cystatin C and lactoferrin were among the most highly expressed antibacterial proteins in pAF. Cystatin C and lactoferrin were confirmed by ELISA to be present in pAF along with lysozyme. Immunoprecipitation of lactoferrin and lysozyme reduced, but did not abolish the antibacterial activities of pAF. Conclusion: Our data demonstrate that pAF maintains antibacterial activity via the preservation of antibacterial proteins against a broad spectrum of wound-associated pathogens.
MeSH term(s)	Amniotic Fluid/metabolism ; Anti-Bacterial Agents/metabolism ; Antimicrobial Cationic Peptides/metabolism ; Humans ; Lactoferrin/metabolism ; Microbial Sensitivity Tests ; Muramidase/metabolism ; Peptides/metabolism ; Pseudomonas aeruginosa/growth & development ; Staphylococcus aureus/growth & development
Chemical Substances	Anti-Bacterial Agents ; Antimicrobial Cationic Peptides ; Peptides ; Muramidase (EC 3.2.1.17) ; Lactoferrin (EC 3.4.21.-)
Language	English
Publishing date	2019-03-01
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1479-5876
ISSN (online)	1479-5876
DOI	10.1186/s12967-019-1812-8
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Article ; Online: Acellular human amniotic fluid protects the ischemic-reperfused rat myocardium.

Lee, Young Sook / Javan, Hadi / Reems, Jo-Anna / Li, Ling / Lusty, Jessica / Schaaf, Christine I / Pierce, Jan / Phillips, John D / Selzman, Craig H

American journal of physiology. Heart and circulatory physiology

2022 Volume 322, Issue 3, Page(s) H406–H416

Abstract: Amniotic products are potent immunomodulators used clinically to repair tissue injury. Little information exists regarding the potential of cell-free human amniotic fluid (hAF) to treat cardiovascular disease. Herein, we sought to determine the influence ...

Abstract	Amniotic products are potent immunomodulators used clinically to repair tissue injury. Little information exists regarding the potential of cell-free human amniotic fluid (hAF) to treat cardiovascular disease. Herein, we sought to determine the influence and efficacy of acellular hAF on myocardial ischemia-reperfusion injury. Processed hAF was obtained from volunteer donors at the time of elective caesarean section and manufactured using proprietary methods. Left anterior descending coronary artery ligation was performed on rats for 60 min. Thirty minutes after release and reperfusion, either saline or hAF was injected intramyocardially. Serial echocardiography revealed that compared with saline-injected rats, hAF animals maintained their ejection fraction and did not adversely remodel through the 4-wk period. This preserved ventricular function correlated with decreased infarct size, less fibrosis, and reduced expression of cytokines and infiltrating inflammatory cells. Comparative arrays of different donor hAF lots confirmed the presence of a wide array of immunomodulatory and host-defense proteins. The observed functional cardioprotection was furthermore evident when given intravenously and across multiple hAF donors. In conclusion, our data demonstrate, for the first time, the cardioprotective effect of acellular hAF on myocardial injury. These observations spanned across diverse donors and likely result from the mixture of a plethora of naturally produced cytokines, chemokines, and immune-modulating proteins rather than a single, defined mechanistic culprit. The ubiquitous availability of hAF as a cell-free solution further suggests its potential for widespread adoption as a therapy for myocardial ischemia-reperfusion injury.
MeSH term(s)	Amniotic Fluid/chemistry ; Animals ; Cardiotonic Agents/analysis ; Cardiotonic Agents/therapeutic use ; Cytokines/genetics ; Cytokines/metabolism ; Female ; Humans ; Immunologic Factors/analysis ; Immunologic Factors/therapeutic use ; Male ; Myocardial Reperfusion Injury/drug therapy ; Myocardium/metabolism ; Rats ; Rats, Sprague-Dawley ; Ventricular Function
Chemical Substances	Cardiotonic Agents ; Cytokines ; Immunologic Factors
Language	English
Publishing date	2022-01-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	603838-4
ISSN	1522-1539 ; 0363-6135
ISSN (online)	1522-1539
ISSN	0363-6135
DOI	10.1152/ajpheart.00331.2021
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Article ; Online: Processed human amniotic fluid retains its antibacterial activity

Yong Mao / Jan Pierce / Anya Singh-Varma / Michael Boyer / Joachim Kohn / Jo-Anna Reems

Journal of Translational Medicine, Vol 17, Iss 1, Pp 1-

2019 Volume 9

Abstract: Abstract Background Human amniotic fluid (AF) contains numerous nutrients, trophic factors and defense proteins that provide a nurturing and protective environment for fetal development. Based on reports that AF has antibacterial, anti-inflammatory and ... ...

Abstract	Abstract Background Human amniotic fluid (AF) contains numerous nutrients, trophic factors and defense proteins that provide a nurturing and protective environment for fetal development. Based on reports that AF has antibacterial, anti-inflammatory and regenerative properties, we designed a novel method to process AF for use in clinical care. Methods Six randomly selected lots of processed AF (pAF) were examined to determine whether they retained their antibacterial activity against a panel of wound-associated pathogens E. faecium, S. aureus, K. pneumoniae, A. baumannii, P. aeruginosa, and E. aerogenes (ESKAPE). To identify proteins in pAF that might be responsible for its antibacterial activity, three different lots of pAF were analyzed with quantitative cytokine arrays that consisted of 400 unique human proteins. One protein identified by microarrays, lactoferrin, and a second prominent antibacterial protein that was not identified by microarrays, lysozyme, were examined by depletion experiments to determine their contribution to the antibacterial activity of pAF. Results All six lots of pAF exhibited antibacterial activity against ESKAPE microorganisms, especially against the pathogens predominately found in chronic wounds (i.e. S. aureus and P. aeruginosa). Thirty-one of the peptides on the microarray were annotated as having antibacterial activity and 26 of these were detected in pAF. Cystatin C and lactoferrin were among the most highly expressed antibacterial proteins in pAF. Cystatin C and lactoferrin were confirmed by ELISA to be present in pAF along with lysozyme. Immunoprecipitation of lactoferrin and lysozyme reduced, but did not abolish the antibacterial activities of pAF. Conclusion Our data demonstrate that pAF maintains antibacterial activity via the preservation of antibacterial proteins against a broad spectrum of wound-associated pathogens.
Keywords	Amniotic fluid ; Antibacterial ; ESKAPE ; Immunoprecipitation ; Medicine ; R
Subject code	572
Language	English
Publishing date	2019-03-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
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Article ; Online: A multicenter evaluation of heterogeneity in cellular therapy processing laboratory procedure times to assess workload capacity.

Thibodeaux, Suzanne R / McKenna, David H / Szczepiorkowski, Zbigniew M / Fontaine, Magali J / Kelley, Linda / Reems, Jo-Anna / Young, Pampee P

Transfusion

2020 Volume 60, Issue 8, Page(s) 1811–1820

Abstract: Background: Growth in size and complexity of clinical hematopoietic progenitor cell (HPC) transplant programs necessitates parallel increases in cellular therapy laboratory (CTL) workload. Typically individually developed, HPC product processing is ... ...

Abstract	Background: Growth in size and complexity of clinical hematopoietic progenitor cell (HPC) transplant programs necessitates parallel increases in cellular therapy laboratory (CTL) workload. Typically individually developed, HPC product processing is labor and time intensive. Variation in procedure type and numbers across CTLs complicates direct comparisons, and benchmark data are not readily available. Study design and methods: Studies were undertaken at seven CTLs. Transplant volume and staff numbers were determined. Staff recorded time performing tasks broken down into steps: paperwork, product acceptance, transport/infusion, processing, and cryopreservation. Times were added to obtain total times for 15 common CTL procedures. Results: Annual transplant volume ranged from 53.4 to 463.2, with products processed by a range of 2 to 10 dedicated CTL staff. Paperwork time constituted 23.7% to 62.3% total time; product processing time accounted for 1.8 (for National Marrow Donor Program product receipt) to 62.6% (for red blood cell reduction of allogeneic HPC products from bone marrow) of total processing time. Mean time for 15 procedures ranged from 1.27 to 8.28 hours (standard deviation range, 0.35-2.71 hr). Mean time for products from bone marrow versus peripheral blood was 6.6 ± 2.0 versus 5.5 ± 1.1 hours (p = 0.02). Cryopreservation (6.5 ± 1.6 vs. 4.4 ± 0.85 hr; p < 0.01) and manipulation (6.4 ± 1.5 vs. 4.4 ± 0.85 hr; p < 0.01) added time. Conclusion: CTL procedures are time intensive, with wide intra- and inter-CTL variation. Paperwork accounted for substantial portion of total time across procedures. Bone marrow source, cryopreservation, and manipulation contributed to longer times. These findings provide concrete data on which to build regarding CTL workload capacity.
MeSH term(s)	Allografts ; Bone Marrow Transplantation ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Laboratories, Hospital ; Workload
Language	English
Publishing date	2020-07-12
Publishing country	United States
Document type	Evaluation Study ; Journal Article ; Multicenter Study
ZDB-ID	208417-x
ISSN	1537-2995 ; 0041-1132
ISSN (online)	1537-2995
ISSN	0041-1132
DOI	10.1111/trf.15899
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Article ; Online: Multicenter evaluation of the IL-3-pSTAT5 assay to assess the potency of cryopreserved stem cells from cord blood units: The BEST Collaborative study.

Trépanier, Patrick / Fournier, Diane / Simard, Carl / Fontaine, Magali J / Stroncek, David / Takanashi, Minoko / McKenna, David / Schwartz, Joseph / Tanhehco, Yvette C / Reems, Jo-Anna / Torrents, Silvia / Kogler, Gesine / Liedtke, Stefanie / Giroux, Martin / Holovati, Jelena L / Louis, Isabelle / Prasath, Arun / Pineault, Nicolas / Bazin, Renée

Transfusion

2022 Volume 62, Issue 8, Page(s) 1595–1601

Abstract: Background: The IL-3-pSTAT5 assay, a new, rapid, and standardized flow-cytometry-based assay may compensate for several limitations of the colony-forming unit (CFU) assay typically used for stem cell potency assessments of cord blood units (CBU). We ... ...

Abstract	Background: The IL-3-pSTAT5 assay, a new, rapid, and standardized flow-cytometry-based assay may compensate for several limitations of the colony-forming unit (CFU) assay typically used for stem cell potency assessments of cord blood units (CBU). We performed an inter-laboratory evaluation of the performance of this new assay. Study design and methods: This Biomedical Excellence for Safer Transfusion (BEST) Collaborative multicenter, international study included 15 participants from public cord blood banks (CBBs), CBB-supporting research laboratories, and stem cell laboratories. To perform the IL-3-pSTAT5 assay, participating centers received reagents, instructions, and 10 blind CBU samples, including eight normal samples and two samples exposed to a transient warming event. We measured inter-laboratory agreement qualitatively (proportion of correctly classified samples) and quantitatively (coefficient of variation [CV], correlation coefficients, receiver operating characteristics (ROC) curve, and intraclass correlation coefficient [ICC]). Results: The qualitative agreement was 97.3% (i.e., 107/110; Fleiss' kappa = 0.835). The average CV on a per-sample basis was 11.57% among all samples, 8.99% among normal samples, and on a per-center basis was 9.42% among normal samples. In a correlation matrix that compared results across centers, the mean Pearson's correlation coefficient was 0.88 (standard deviation = 0.04). The ICC was 0.83 (95% confidence interval = 0.68-0.95). The area under the curve (AUC) from the ROC curve was 0.9974. Discussion: Excellent qualitative and quantitative agreement was exhibited across laboratories. The IL-3-pSTAT5 assay may therefore be implemented in flow cytometry laboratories to rapidly and reliably provide standardized measures of stem cell potency in CBUs.
MeSH term(s)	Blood Banking/methods ; Colony-Forming Units Assay ; Fetal Blood ; Humans ; Interleukin-3 ; STAT5 Transcription Factor/metabolism ; Stem Cells
Chemical Substances	Interleukin-3 ; STAT5 Transcription Factor
Language	English
Publishing date	2022-06-30
Publishing country	United States
Document type	Journal Article ; Multicenter Study ; Research Support, Non-U.S. Gov't
ZDB-ID	208417-x
ISSN	1537-2995 ; 0041-1132
ISSN (online)	1537-2995
ISSN	0041-1132
DOI	10.1111/trf.16997
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