Cord Blood Banking Research Articles

Establishing the ethical and regulatory frame work for a clinically compliant ipsc cell line bank; from qa to donor reconsent.

Background & Aim A global network of iPSC banks would create a strong future basis for new cellular therapies, providing GMP quality clinically complaint cell lines to produce differentiated cells to facilitate research studies through to clinical trials. Cryopreserved Cord Blood (CB) is an excellent starting material for iPSC production, however setting up these banks requires clear and transparent reconsent of donors along with a strong QA and QC framework. We aim to provide a clear ethical pathway towards the creation of such a resource, along with a dynamic follow up and reconsent process of donors from the public BMDI Cord Blood Bank. Methods, Results & Conclusion Review of the literature and a round-table workshop with Ethics and Legal experts have identified the key issues to be resolved in order to obtain consent to make iPSC lines from CB donors. Regulatory challenges that would affect global acceptance of any cell lines, along with the Quality framework and the standards required to operate as part of a global network, are being met by working in collaboration with bodies such as the Global Alliance for iPSC Therapies (GAiT) and participating in an iPSC Quality Assessment Round. Developing the mechanism for reconsenting donors and increasing their understanding to allow fully informed consent, in addition to establishing ethical guidelines for confidentiality, have presented challenges. CB donor letters and information sheets have been developed and Ethics approvals have been sought and processed by an Institutional HREC, prior to submission for government approval. New issues of whole genome sequencing and the relevant donor safeguards and protections have been considered with input from clinical genetics services, including the rights and information flow to donors about what happens to the cells after banking and commercialization aspects. Freedom to operate and GMP manufacture and oversight is currently under investigation. This work is continuing and the ongoing success of many of these processes has confirmed feasibility and utility of using banked CB to produce clinical–grade iPSC lines for potential cellular therapies. By facilitating clinical research, we can improve Australia's position in the clinical trials space, providing a standardised and characterised resource that could lead to eventual new cell therapies that will be available to Australian and international communities.

Production of iPSCs from a small volume of cryopreserved human umbilical cord blood buffy coat under “gmp-compliant” conditions

Background & Aim An iPSC bank derived from cord blood selected on the basis of human leukocyte antigens (HLA) requires the development and validation of protocols to generate iPSC lines under good manufacturing practice (GMP) conditions. Working towards this goal, we aim to create clinically compliant GMP grade iPSCs from small volumes of banked cord blood (CB) buffy coat samples for potential clinical use. Methods, Results & Conclusion We have established a protocol to generate iPSC lines from a minimum of 50 µl of cryopreserved CB buffy coat using defined medium and reagents manufactured under GMP. This allows us to use only segments or pilot tube of the stored CB unit without compromising the whole CB unit. Thawed CB cells were washed and an erythroblast cell population expanded in Animal Component Free (ACF) erythroblast growth medium prior to reprograming. Up to 0.11% reprogramming efficiency was obtained. iPSC colonies formed on Vitronectin coated plates were manually picked and passaged on the same matrix in defined Xeno-free E8 Flex medium. Karyotype integrity of the generated iPSC lines was confirmed with SNP Illumina Infinium CoreExome-24 v1.1 array at 0.50Mb resolution. Flow cytometry was used to show that iPSCs expressed the known stemness markers SSEA-3, SSEA-4, TRA-1-81 and TRA-1-60R but lacked expression of SSEA-1. In vitro monolayer differentiation experiments indicated that CB derived iPSCs could differentiate into cells representing the 3 germline lineages: beating cardiac cells (mesoderm), neuronal cells (ectoderm) and endodermal precursor cells. All derived cell types demonstrated the appropriate lineage markers by FACS or immunochemistry. The established iPSC line was passaged for >45 times under current culture conditions to demonstrate the stability of karyotype integrity and pluripotency potential of the cells, consistent with the earlier passages. Differentiation into cardiomyocytes indicates that differentiation capacity is similar to the earlier passages. We have demonstrated that GMP-compliant iPSC lines can be reproducibly and stability generated from small volumes of cryopreserved umbilical cord blood. By using the protocols established through this work we will create clinically compliant cell lines that will facilitate clinical research and the creation of an Australian CB-derived iPSC “haplobank” for cellular therapies.

Parathyroid Tissue Cryopreservation: Does the Storage Time Affect Viability and Functionality?

Parathyroidectomy is a standard practice to treat recurrent or persistent hyperparathyroidism. However, this can lead to the onset of hypoparathyroidism, treatable with the autotransplantation of parathyroid tissue (PT). Tissue can be transplanted immediately after parathyroidectomy or cryopreserved and transplanted only in case of necessity. Since 2011, the Cord Blood Bank and Cardiovascular Tissue Bank of Emilia-Romagna has been storing PT for potential autologous transplantation. To date, there are highly variable data about the viability and function of PT after thawing. However, it is not clear if the PT quality is affected by different cryopreservation protocols and/or by the storage time. The aim of this study was to assess the ex vivo function and viability of the PTs of ten patients stored in the Bank. Tissue morphology was evaluated before and after cryopreservation through histological investigations. PT function was analyzed by assessing the ability of cryopreserved PT to synthesize and secrete parathyroid hormone (PTH) in response to different calcium concentrations. Moreover, viability and function were also investigated on tissue-isolated cells in culture. These data show that tested tissues appear to be viable and able to produce PTH even after 5 years of storage, and the histological architecture is well preserved.

Reducing ethnic disparity in access to high-quality HLA-matched cord blood units for transplantation: analysis of the Canadian Blood Services' Cord Blood Bank inventory.

Launched in 2013, Canadian Blood Services' Cord Blood Bank (CBS' CBB) has built a high-quality, ethnically diverse cord blood repository that aims to reduce ethnic disparity in accessing suitable units for transplantation. As of December 2016, 2000 units have been banked. The self-reported maternal ethnicity was 58% non-Caucasian. Overall, 26% of units were classified as multi-ethnicity with Caucasian (84%) most frequently observed in combination with Asian, First Nations (predominant indigenous peoples in Canada south of the Arctic Circle), or African ethnicity. Utilization scores that incorporate total nucleated and CD34+ cell counts in the CBS' CBB were associated with greater likelihood of utilization compared with the international inventory of units (p < 0.05). The distribution of utilization scores was similar for Caucasians compared with non-Caucasians (p < 0.05). Using HLA genotypes of cord blood units and their mothers, we determined probable ethnic assignments for each haplotype using HaploStats (National Marrow Donor Program). Significant increases in HLA-match likelihoods are predicted for all ethnicities as the inventory grows to its target of 10,000 units and the gap in HLA-match likelihoods for Caucasian and non-Caucasian patients progressively declines. The CBS' CBB inventory is predicted to have high HLA-matching likelihoods across a broad spectrum of ethnic groups, improving access to high-quality stem cell products for all patients.

Repurposing the public cord blood bank inventory in Korea to enhance cord blood use

To enhance public cord blood (CB) use, we examined the current status of CB banking and tried to suggest revision of the banking standard. We retrospectively analyzed the use of stored public CB units between 2011 and 2016 using data from the CB information center in Korea. A total of 19,871 CB units were registered, and 363 units were selected for transplantation. The transplanted CB units contained significantly higher numbers of CD34+ cells than the average numbers in the stored CB units (5.5 × 10^6 vs. 3.2 × 10^6, p < 0.01). They also contained more total nucleated cells (TNCs) than the average of the stored CB units (13.7 × 10^8 vs. 10.7 × 10^8, p < 0.01). Only 49% of the stored CB units contained>10 × 10^8 TNCs, while 81% of the units transplanted contained >10 × 10^8 TNCs. The average length of cryopreservation of the transplanted CB units was 4.58 years and 95% of them had been stored for less than 10 years. During the study period, 18,763 CB units were requested for research, but only 5,888 were released. This discrepancy was mostly due to errors in regulatory and/or networking elements of the CB supply system. The data suggest that preserving CB units for less than 10 years and increasing the required minimum TNC count to 10 × 10^8 would produce an inventory containing units that were more useful for CBT. CB units that did not meet the requisite quality standards could be used for research, and systems for their fair distribution to researchers are needed.

Забезпечення публічного інтересу у сфері донорства крові

Забезпечення публічного інтересу у сфері донорства крові

Adapting Cord Blood Collection and Banking Standard Operating Procedures for HLA-Homozygous Induced Pluripotent Stem Cells Production and Banking for Clinical Application.

In this article, we will discuss the main aspects to be considered to define standard operation procedures (SOPs) for the creation of an induced pluripotent stem cell (iPSC) bank using cord blood (CB)—or similar cell type—bank guidelines for clinical aims. To do this, we adapt the pre-existing SOP for CB banking that can be complementary for iPSCs. Some aspects of iPSC manufacturing and the particular nature of these cells call for special attention, such as the potential multiple applications of the cells, proper explanation to the donor for consent of use, the genomic stability and the risk of genetic privacy disclosure. Some aspects of the iPSC SOP are solidly established by CB banking procedures, other procedures have good consensus in the scientific and medical community, while others still need to be further debated and settled. Given the international sharing vocation of iPSC banking, there is an urgent need by scientists, clinicians and regulators internationally to harmonize standards and allow future sample interchange between many iPSC bank initiatives that are springing up worldwide.

Cord Blood Banking: Antenatal Care Provider's Roles and Responsibilities.

Background Umbilical Cord Blood (UCB) banking done either for private storage or for donation to public cord blood banks involves active participation of obstetricians. Counseling the expectant parents, providing them with unbiased and balanced information, and collecting the UCB with diligence confer a lot of social as well as moral responsibility upon obstetricians. This makes it even more important that the obstetricians in current practice stay well-informed and updated with UCB collection and its storage guidelines. The present study was conducted to assess the current status of obstetricians about UCB banking in terms of their awareness, attitude, and expectations from it. Materials and Methods A cross-sectional study was conducted across three hospitals. A self-administered 22-item questionnaire was given to obstetricians to assess their awareness, attitude, and expectations about UCB banking. Finally, 154 completed questionnaires were analyzed using SPSS software (version 15.0). The awareness, attitude, and expectations were assessed and reported as primary endpoints and the self-rated knowledge levels, and sources of information were reported as secondary endpoints. Results Overall, the awareness was poor, but the attitude was favorable for UCB banking amongst obstetricians. Around 74% felt that obstetricians must be well-informed about UCB banking-related counseling and collection protocols. However, 55% felt it to be an additional burden for the obstetrician, and 57% believed that financial compensation must be given to obstetricians involved with cord blood collection procedures. The majority remained unclear about their expectations from UCB banking. The self-rated knowledge was poor and very poor for 75% obstetricians. 89.6% derived their information from representatives of private cord blood companies. Conclusion Although poor in awareness levels, obstetricians possessed a favorable attitude towards UCB banking. Continuing medical education needs to focus more on such current issues of public importance to keep professionals updated. This is one way to minimise percolation of wrong facts and figures by the industries with conflicting interest to the healthcare providers.

Model Criteria for Regulation of Cord Blood Banks and Cord Blood Banking: Adopted by the Cord Blood Association, Board of Directors, January 29, 2019.

Model Criteria for Regulation of Cord Blood Banks and Cord Blood Banking: Adopted by the Cord Blood Association, Board of Directors, January 29, 2019.

Long-Overdue Guidelines for the Cord Blood Banking Community.

Long-Overdue Guidelines for the Cord Blood Banking Community.

ACOG Committee Opinion No. 771: Umbilical Cord Blood Banking.

Since the first successful umbilical cord blood transplant in 1988, it has been estimated that more than 35,000 transplants have been performed in children and adults for the correction of inborn errors of metabolism, hematopoietic malignancies, and genetic disorders of the blood and immune system. Two types of banks have emerged for the collection and storage of umbilical cord blood: 1) public banks and 2) private banks. The benefits and limitations of public versus private umbilical cord blood banking should be reviewed with the patient because they serve different purposes. This patient discussion also should include the concept of autologous and allogeneic use of umbilical cord blood. Umbilical cord blood collected from a neonate cannot be used to treat a genetic disease or malignancy in that same individual (autologous transplant) because stored cord blood contains the same genetic variant or premalignant cells that led to the condition being treated. There is no current evidence to support the use of an autologous umbilical cord blood sample in regenerative medicine. Patients should be made aware of the quality control and regulatory organizations that provide oversight for the process of umbilical cord collection and storage. Umbilical cord blood collection should not compromise obstetric or neonatal care or alter routine practice of delayed umbilical cord clamping with the rare exception of medical indications for directed donation. Therefore, it is important to inform patients that the medical condition of the woman or neonate may prevent adequate umbilical cord blood collection. This document is updated with a statement that the routine use of private cord blood banking is not supported by available evidence and that public banking is the recommended method of obtaining cord blood. In addition, the importance of contribution from all ethnicities and races to public banks is highlighted.

ACOG Committee Opinion No. 771 Summary: Umbilical Cord Blood Banking

Since the first successful umbilical cord blood transplant in 1988, it has been estimated that more than 35,000 transplants have been performed in children and adults for the correction of inborn errors of metabolism, hematopoietic malignancies, and genetic disorders of the blood and immune system. Two types of banks have emerged for the collection and storage of umbilical cord blood: 1) public banks and 2) private banks. The benefits and limitations of public versus private umbilical cord blood banking should be reviewed with the patient because they serve different purposes. This patient discussion also should include the concept of autologous and allogeneic use of umbilical cord blood. Umbilical cord blood collected from a neonate cannot be used to treat a genetic disease or malignancy in that same individual (autologous transplant) because stored cord blood contains the same genetic variant or premalignant cells that led to the condition being treated. There is no current evidence to support the use of an autologous umbilical cord blood sample in regenerative medicine. Patients should be made aware of the quality control and regulatory organizations that provide oversight for the process of umbilical cord collection and storage. Umbilical cord blood collection should not compromise obstetric or neonatal care or alter routine practice of delayed umbilical cord clamping with the rare exception of medical indications for directed donation. Therefore, it is important to inform patients that the medical condition of the woman or neonate may prevent adequate umbilical cord blood collection. This document is updated with a statement that the routine use of private cord blood banking is not supported by available evidence and that public banking is the recommended method of obtaining cord blood. In addition, the importance of contribution from all ethnicities and races to public banks is highlighted.

Extracellular vesicles: Squeezing every drop of regenerative potential of umbilical cord blood

Collection, cryopreservation and transplantation of umbilical cord blood (UCB)-derived cells have become a popular option for regenerative medicine, not limited to the transplantation of hematopoietic cell progenitors only. Indeed, increasing evidence shows that extracellular vesicles (EV), which include a heterogeneous pool of membranous structures secreted by the vast majority of cells, can serve as powerful tools for cell-free therapy due to precise multifunctional molecular cargoes. In this issue, Hu et al. [1] described that EV extracted from UCB (UCB-EV) ameliorate bone loss in senile osteoporotic mice and promote in vitro osteoblast differentiation of bone marrow-derived Mesenchymal Stromal Cells through miR-3960-mediated signaling. These results envision the capability of UCB-EV of priming multipotent stem cells toward the osteogenic cell lineage and interfering on bone resorption processes. Although processing and manufacturing of EV-based products have to further develop major issues, we foresee that EV will soon become new therapeutic products supplied by UCB banks for treating human diseases, including bone-related conditions.

An objective flow cytometry method to rapidly determine cord blood potency in cryopreserved units.

Cord blood banks have to determine the regenerative potential of cord blood units (CBUs) on a representative sample of the cryopreserved product before release to the transplant center. Potency can be measured by using a colony-forming unit (CFU) method, which delays the release of CBU by 7 to 14 days. To accelerate CBU qualification, we have developed a rapid method to assess the response of CD34 cells to interleukin (IL)-3. Flow cytometry was used to measure IL-3-induced STAT5 phosphorylation within CD34-cells. This IL-3 test was compared to the CFU method, as well as the aldehyde dehydrogenase (ALDH) enzyme-based assay. Ten cryopreserved CBUs were analyzed for their contents in CD34 and CD45 viable cells, total CFUs, ADLHbright cells, and IL-3-responsive CD34+ cells. Extreme and mild warming event scenarios were simulated on CBUs and used as poor-quality samples. Segments, tubes, and bags from five CBUs were compared for their potency using IL-3 and CFU methods. The IL-3 test was accurate in identifying the samples handled following standard operating procedures and those subjected to extreme warming events. Based on these results, a threshold of 55% of IL-3-responsive CD34 cells was established to identify good-quality samples. The IL-3 test was also the most sensitive to detect samples subjected to milder warming events. Our new method for determining CBU functionality is rapid, unbiased, and robust. The IL-3 test described herein fulfills the requirements for validation, and we intend to implement this method in our cord blood bank facility.

Contextual factors influencing donor recruitment and cord blood collection: perspectives of frontline staff of the Canadian Blood Services' Cord Blood Bank.

Umbilical cord blood (CB) is an important source of hematopoietic stem cells that are used to treat blood- and immune-system disorders. Public CB banks aim to build inventories with high-quality CB units to meet healthcare needs. While research has noted the influence of broader contextual factors on donor recruitment and CB collection processes, to date, no published study has identified the specific contextual factors and challenges to donor recruitment and CB collection. This paper addresses this gap in the literature. A qualitative case study focusing on donor recruitment and CB collection processes was conducted to identify the contextual factors influencing these processes. This paper reports the findings from in-depth, semi-structured interviews conducted with 15 frontline staff of the Canadian Blood Services' Cord Blood Bank. Interview data were analyzed using inductive interpretive methods to identify the contextual conditions and factors that influence recruitment and collection. Frontline staff described various social factors that influenced and challenged the processes of donor recruitment and CB collection. These were categorized into four overlapping contexts: birthing context, hospital context, CB bank organizational context, and sociocultural context. Consideration of social context is necessary in order to effectively address the factors and challenges that influence the successful development of high-quality CB inventories, and to guide resource allocation. Further examination of contextually-rooted factors and their interactions is necessary to optimize donor recruitment and CB collection processes.

Isolation, Culture and Biological Characteristics of Endothelial Progenitor Cells from Cryopreserved Umbilical Cord Blood

To establish a novel method to isolate endothelial progenitor cells（EPC） from cryopreserved umbilical cord blood (cryoUCB), to investigate the biological characteristics of EPC and to improve the rate of EPC obtained from cryoUCB. Twelve cryoUCB samples during 2000 to 2001 years were collected from allogeneic cord blood bank, cryoUCB was thawed rapidly in a water bath at 37 ℃, total nucleated cells (TNCs) were washed by phosphate-buffered saline (PBS). TNCs were seeded onto fibronectin-coated dishes to isolate EPC. Flow cytometry and immunofluorescence were used to identify EPC. The function of EPC was identified in vitro, such as the incorporation of Dil-Ac-LDL and FITC-UEA-I, the formation of capillary-like structure in matrigel, and the release of VEGF by ELISA. One to five cluster of cobble stone-like cells appeared at 2-3 weeks after seeding. Flow cytometric analysis showed that positive rates of CD31, CD34, CD144, and VEGFR (CD309) were（92.91±5.20）%, （30.0±23.27）%, （88.55±3.83）% and （67.21±12.12）% in passage 1 to passage 3 of EPC. EPC could uptake Dil-Ac-LDL and FITC-UEA-I, form capillary-like network on Matriget and release VEGF. EPC had been successfully isolated from cryopreserved umbilical cord blood by this method with high stability and reproducibility. EPC can be obtained in 85% frozen umbilical cord blood. This method may lay a foundation to supply abundant EPC for clinical application.

The Future State of Newborn Stem Cell Banking.

Newborn stem cell banking began with the establishment of cord blood banks more than 25 years ago. Over the course of nearly three decades, there has been considerable evolution in the clinical application of stem cells isolated from newborn tissues. The industry now finds itself at an inflection point as personalized medicine and regenerative medicine continue to advance. In this review, we summarize our perspective on newborn stem cell banking in the context of the future potential that stem cells from perinatal tissues are likely to play in nascent applications. Specifically, we describe the relevance of newborn stem cell banking and how the cells stored can be utilized as starting material for the next generation of advanced cellular therapies and personalized medicine.

Time dependent risk of cytomegalovirus infection in Japan.

Cytomegalovirus (CMV), a major cause of congenital infections, has high morbidity and mortality rates associated with it. However, a decline in the proportion of anti-CMV antibody-positive individuals has been observed. The present study aimed to quantify the time-dependent transmission dynamics of CMV infection in Japan by analysing the seroepidemiological datasets for pregnant women collected from five cord blood banks from 1996 to 2009. By employing a mathematical model and using the maternal age distribution of child births from the census data, we computed the seroprevalence among the pregnant Japanese women as a function of time. A decreasing trend was observed for the force of infection, i.e. the rate at which susceptible individuals are infected, which decreased from 0.04 to 0.03 (/year) over the period from 1996 to 2009. While the total number of births has steadily declined in Japan over time, the estimated number of live births at risk of CMV infection has increased over time. Our data reveal that in 2009 in Japan, at least 0.3 million women may have been at risk of contracting a CMV infection during the perinatal period. Moreover, about 2,726 congenital CMV infections were expected to have occurred in 2009. The average age at infection has already reached the child bearing age, and it must be noted that the age at infection can be elevated even more, reaching close to 30 years old which is the ongoing mean age at child delivery. It must be remembered that, if vaccine can become one of the options for the control of CMV in the future, the vaccination can lead to further elevation of age at infection, which may coincide with further elevation of mothers' age of delivery in Japan.

A Study to assess the effectiveness of Self Instructional Module on the knowledge regarding cord blood banking among staff nurses in selected hospitals, Tumkur

A Study to assess the effectiveness of Self Instructional Module on the knowledge regarding cord blood banking among staff nurses in selected hospitals, Tumkur

Kordon Kanı Bankacılığı ve Etik

Kordon kanı, hematopoetik kök hücrelerin toplanması ve nakledilmesi için bir kaynak olarak kabul edilmektedir. Kordon kanında bulunan kök hücreler, hem vericinin kendisi hem de kardeş ve akrabaları için oldukça yüksek doku uyumu göstermektedir. Kordon kanının tıbben kullanılabilir ve saklanabilir olması yarar sağlamakla birlikte, toplanmasından kullanımına kadar geçen süre içerisinde etik sorunların oluşumuna yol açmaktadır. Etik sorunlar özerklik, yarar sağlama-zarar vermeme, bilgilendirme gibi etik ilkelerin zedelenmesine neden olabilmektedir. Bu çalışmada, kordon kanı ve bankacılığının oluşturduğu etik sorunlara dikkat çekilmesi amaçlanmıştır.