Quality Of Umbilical Cord Blood Research Articles

Evaluation of factors affecting total nucleated cells in umbilical cord blood collected for the Calabria Cord Blood Bank.

Total nucleated cell (TNC) count is the most important biological feature to consider in assessing the quality of umbilical cord blood (UCB) for hematopoietic stem cell (HSC) transplantation. Certain obstetric factors have been reported to increase TNC count in UCB units collected for transplantation. The aim of our study was to analyze how various maternal, neonatal and obstetric factors affected TNC count in the UCBs we collected for our cord blood bank in southern Italy. We performed a retrospective analysis of 634 medical records of UCBs collected by Calabria Cord Blood Bank (CCBB), Reggio Calabria, Italy, between January 1, 2010, and December 31, 2016. We analyzed various maternal, neonatal and obstetric variables factors and related this factor with the characteristic of TNC. We found that the average number of TNCs was significantly greater in vaginal delivery than in caesarean delivery. We also found that TNCs were higher in the 40th week of pregnancy and when Apgar 1' scores were ≤9. The effect of a newborn's gender was less evident on TNC count. Knowledge of factors predictive of a higher TNC count would help cord blood banks more efficiently identify donors likely to yield high-quality UCBs for transplantation.

Predictors of cord blood unit cell content in a volume unrestricted large series collections: a chance for a fast and cheap multiparameter selection model

BackgroundCord blood plays a very important role in stem cell transplantation and therapy with an emerging implication also in regenerative medicine. The number of cells available in a single cord blood unit (CBU), in particular, the CD34+ and total nucleated cell (TNC) content influences the transplantation clinical outcome. We analysed a very large series of CBUs, collected for private banking without any specific volume restriction, to deeply investigate the best predictors of cord blood stem cells content.MethodsMaternal and neonatal clinic laboratory data of a total 2583 UCBs were obtained from the InScientiaFides cord blood bank based in Republic of San Marino. Univariate and multivariate analysis were conducted to better interpret the data and to build a predictive model to select, the CBU with high CD34+ content.ResultsOur univariate analysis shows that seasonality and the geographical area affects the quality of umbilical cord blood. Gestational age, babie’s gender and birth weight have a positive correlation with CB TNC content. The babie’s birth weight affects positively also CD34+ content and CBU volume while the cesarean delivery affect the CB volume only. Our predictive model, based on multivariate analysis, shows that male babie’s, gestational age lower to 39 weeks, cesarean delivery and CBUs with a content of TNC higher than 3.44 × 108 (group A) have a significant higher CD34+ content than group B (female babie’s, gestational age higher than 39 weeks and vaginal delivery). The group A have a 37.5% of CBUs with a concentration of CD34+ > 2 × 106, while no CBUs with high concentration of CD34+ were detect in group B.ConclusionThis study, conducted on a very large series of CBUs without any specific volume constraint, highlighted the prenatal and maternal factors that significantly influence the quality of the CBU collected. Specifically, it highlights that volume is not the best predictor of CD34+ CBU content; for this reason it cannot be taken into consideration alone for the analysis of the collected samples. Our final aim is to identify relevant factors, immediately available, that help to choice UCB with high CD34+ cell content, especially in simultaneous deliveries.

No title available] - PubMed

To study the impact of anticoagulant to the quality of umbilical cord blood (UCB). 6060 cord blood units (CBUs) were classified into five groups, such as 28 ml: (10-29) ml, 28 ml: (30-69) ml, 28 ml: (70-109) ml, 28 ml: (110-150) ml and 28 ml: (>150) ml according to volume ratio of anticoagulant and CBVs. The count of pre-cryopreservation total nucleated cell (pre-TNC), the viability of nucleated cell (VNC), the amount of CFU-GM and the ratio changes of CD34+ were evaluated and analyzed statistically. It was found that pre-TNC increased with the growth of volume of CBUs (r=0.9937) under the certain volume of antico-agulant, and the TNC in the minimum UCB volume group was (2.57±0.89)×108; the VNC grew up with the increasing count viability of volume (r=0.9897), and the average viability of the minimum volume group remained over 95%; the CFU-GM climbed up with the increasing of volume (r=0.9024), and the number of CFV-GM in minimum volume group reached to of 89/×105; CD34+% grew up with the increase of volume of CBUs (r=0.9641), and the ratio was (0.30±0.19)% for the minimum volume group. In certain volume of anticoagulant in collection-bag, pre-TNC, VNC, CFU-GM and CD34+% are all dropped with the decrease of CBUs volume , however, all above-mentioned indexes in the minimun random group still meet the requirement for clinical administration.

Evaluation of factors affecting total nucleated cells in umbilical cord blood collected for the Calabria cord blood bank.

Total nucleated cell (TNC) count is the most important biological feature to consider in assessing the quality of umbilical cord blood (UCB) for haematopoietic stem cell (HSC) transplantation. Certain obstetric factors have been reported to increase TNC count in UCB units collected for transplantation. The aim of our study was to analyze how various maternal, neonatal and obstetric factors affected TNC count in the UCBs we collected for our cord blood bank in southern Italy. We performed a retrospective analysis of 634 medical records of UCBs collected by Calabria Cord Blood Bank (CCBB), between January 1, 2010 and December 31, 2016. We analyzed various maternal, neonatal and obstetric variables factors and related this factor with the characteristic of TNC. We found that the average number of TNCs was significantly greater in vaginal delivery than in caesarean delivery. We also found that TNCs were higher in the 40th week of pregnancy and when Apgar 1' scores were ≤ 9. The effect of a newborn's gender was less evident on TNC count. Knowledge of factors predictive of a higher TNC count would help cord blood banks more efficiently identify donors likely to yield high-quality UCBs for transplantation.

Factors Associated With Umbilical Cord Blood Collection Quality in Japan.

BackgroundUmbilical cord blood (UCB) has become an established alternative source of hematopoietic stem cells with marrow and postmobilization peripheral blood. The presence of a large amount of clots may lead to the deterioration of cord blood quality. To improve UCB quality as a source of hematopoietic stem cells in Japan, we examined factors associated with UCB collection methods from the viewpoint of eliminating the presence of clots.MethodsIn August 2019, we requested the directors of 74 certified facilities to provide information on UCB collection methods in Japan. A total of 46 (62.2%) of them responded with valid information on a total of 2,892 UCB collections. In this study, collected UCB without clots macroscopically was evaluated as a high-quality UCB.ResultsThe 2,891 UCB collections described during the study period were divided to those with (n = 760, 26.3%) and without clots (high quality; n = 2,131, 73.7%). Multivariate analysis revealed single puncture as a factor determining high-quality UCB collection (adjusted odds ratio (ORs): 1.80, 95% confidence interval (CI): 1.3 - 5.4, P = 0.01).ConclusionsSingle puncture is an independent effective factor determining high-quality manual UCB collection in Japan.

Factors Affecting Human Umbilical Cord Blood Quality Before Cryopreservation: The Importance of Birth Weight and Gestational Age.

Background: Umbilical cord blood (UCB) is a rich source of hematopoietic stem cells and is useful for the treatment of blood diseases. The cost of UCB storage is high; thus, it is necessary to evaluate the quality of UCB before collection and cryopreservation. Aim: This study aimed to determine the maternal and neonatal factors that influence UCB before selection for cryopreservation. Materials and Methods: The analysis included 403 processed UCB units. The effects of maternal characteristics including maternal age and delivery method and neonatal factors such as birth weight, gestation duration, and sex on UCB quality were determined based on the collected blood volume, total nucleated cell (TNC) count, and CD34+ cell count. Results: The neonatal birth weight influenced the collected blood volume, TNC count, and CD34+ cell count. Neonates with higher birth weights produced better quality UCB units because of increased collected blood volumes, TNC counts, and CD34+ cell counts. However, an increase in the gestational age from 35 to 41 weeks led to decreases in the collected blood volume and CD34+ cell count. Conclusion: These data may be useful for determining the optimal cord blood units for collection and cryopreservation and for advising pregnant women using private banking services.

Factors Affecting Human Umbilical Cord Blood Quality Prior to Cryopreservation: The Importance of Birth Weight and Gestational Age

Factors Affecting Human Umbilical Cord Blood Quality Prior to Cryopreservation: The Importance of Birth Weight and Gestational Age

Umbilical cord blood quality and quantity: Collection up to transplantation.

Umbilical cord blood (UCB) is an attractive source of hematopoietic stem cells for transplantation in some blood disorders. One of the major factors that influence on transplantation fate is cord blood (CB) cell count, in addition to human leukocyte antigen similarity and CD34+ cell number. Here, we review the factors that could effect on quality and quantity of CBUs. Relevant English-language literatures were searched and retrieved from PubMed using the terms: CB, quality, collection, and transplantation. The numbers of total nucleated cells (TNCs) and CD34+ cells are good indicators of CB quality because they have been associated with engraftment; thereby, whatever the TNCs in a CB unit (CBU) are higher, more likely they led to successful engraftment. Many factors influence the quantity and quality of UCB units that collect after delivery. Some parameters are not in our hands, such as maternal and infant factors, and hence, we cannot change these. However, some other factors are in our authority, such as mode of collection, type and amount of anticoagulant, and time and temperature during collection to postthaw CBUs and freeze-and-thaw procedures. By optimizing the CB collection, we can improve the quantity and quality of UCB for storage and increase the likelihood of its use for transplantation.

Defining Parameters That Can Guide Public Cord Blood Units (CBUs) in Korea

Background: It is well known that Total Nucleated Cell (TNC) and CD34 are the greatest limiting factor in the use of umbilical cord blood (UCB) for transplantation. To enhance high UCB quality, it is important that how the factors will affect the UCB. This study is to identify maternal, neonatal, obstetric factors that influence the suitability for banking and transplantation of UCB units collection.Study design and methods: This study examined 6534 UCB (3712 at the Catholic Hematopoietic Stem Cell Bank and 2822 at the Daegu Fatima Hospital Public Umbilical Cord Bank) collected at two banks from October 2003 to June 2015. The variables were collected from retrospective records at the time of donation. The associations between TNC, CD34+ and variables including maternal age (MA), gestational age (GA), fetal body weight (FBW), time from collection to processing (T), collecting volume (CV), preTNC, and delivery type were analyzed by logistic regression.Results: In our study cohort (n=6534, male 2988, female 2991, unkown 555, all Koreans), the median values of TNC, numbers of CD34+, MA, GA, FBW, T, preTNC, and CV were 9.24úI108/unit (range, 3.02-35.64), 2.0úI106/unit(range, 0.04-29.2), 31.0 years(range, 15-46), 277 days (range, 202-382), 3330g (range, 1740-4970), 19 hours (range, 1-54), 11.69úI108/unit (range, 3.41-50.32), and 83.5ml (range, 26.0-218.2) respectively.In univariate analysis, variables that were associated with high TNC (defined as a TNC of > 9.24úI108/unit) included GA (defined as GA > 277 days) [OR 1.29 (95% CI 1.16-1.42 p < 0.001)], FBW (defined as FBW > 3330g) [OR 1.52 (95% CI 1.37-1.68 p < 0.001)])], CV (defined as > 83.5mL) [OR 2.61 (95% CI 2.36-2.88 p < 0.001)], preTNC [OR 25.45 (95% CI 22.34-29.00 p < 0.001)], and T (defined as T> 19 hours) [OR 0.87 (95% CI 0.79-0.96 p < 0.005)]. Variables that were associated with high CD34+ (defined as a number of CD34+ > 2.0úI106/unit) included MA (defined as MA > 31.0 years) [OR 0.90 (95% CI 0.82-0.99 P=0.036)], GA [OR 0.74 (95% CI 0.67-0.82 p < 0.001)], FBW [OR 1.41 (95% CI 1.27-1.56 p<0.001)], preTNC [OR 3.38 95% CI 3.06-3.74 p < 0.001]], and CV [OR 1.41 (95% CI 1.28-1.60 p<0.001)]In multivariate analysis of TNC, preTNC [OR 20.71 (95% CI 17.87-24.00) p < 0.001]] was the best predictor of followed by normal delivery [OR 1.77 (95% CI 1.48-2.11 P<0.001)], FBW [OR 1.35 (95% CI 1.17-1.56 p<0.001)], CV [OR 1.31 (95% CI 1.13-1.53 p<0.001)], and female gender [OR 1.21 (95% CI 1.05-1.39 p=0.01)]. In multivariate analysis of CD34, was preTNC [OR 3.39 (95% CI 3.00-3.83 p < 0.001)] was the best predictor of followed by FBW [OR 1.41 (95% CI 1.25-1.58 P<0.001)], GA [OR 0.59 (95% CI 0.52-0.66 p<0.001)], MA [OR 0.84 (95% CI 0.75-0.94 p=0.003)], and female gender [OR 0.89 (95% CI 0.78-0.98 p=0.02)].Conclusions: We established referential values of cord blood using large scaled CB units in Korea. In multivariate analysis, maternal/donor characteristics were associated with preTNC, FBW, and gender for both high TNC and CD34+. Our results confirm that is similar values to those reported in previous data. These associations could be used to prioritize donations, collections, optimizing resource utilization and financial modeling in Korean cord blood banks. We are focusing on collection education using the standard operation procedure to facilitate of high cells as well as on more recruits of healthy mothers. DisclosuresNo relevant conflicts of interest to declare.

The impact of fetal and maternal physiologic factors on umbilical cord blood quality as a source of stem cells in Egyptian population.

Umbilical cord blood (UCB) has rapidly become a clinically useful alternative stem cell source. Many variables have been used to evaluate a UCB unit and predict transplant outcomes. The objective of this study was to measure the expression of hematopoietic stem cells in UCB and its relation to certain maternal and neonatal physiologic factors to establish optimum criteria for UCB donor selection. Two hundred UCB units were collected from normal uncomplicated vaginal and cesarean deliveries. Total volume was noted and immediately assessed for total nucleated cell (TNC) count and CD34+ cell concentration. Assessment of maternal and neonatal variables such as mode of delivery, placental weight, baby's birthweight, and sex was made. The volume of the donations ranged from 42.0 to 126 mL, the TNC count ranged from 5 × 10(9) to 28.7 × 10(9) cells/L, and CD34+ cells ranged from 0.03% to 0.62%. There was a significant positive correlation between cord blood volume and cesarean section (p = 0.01) and placental weight (p = 0.02). There was a significant positive correlation with a p value of less than 0.05 between the number of CD34+ cells and UCB volume and TNC. There was no significant difference between the variables and the TNC count. Our study concludes that cord units collected for banking should be obtained by selecting units of larger volumes, of higher TNCs, from female babies with heavy placenta, and from babies delivered via cesarean section.

Postthaw characterization of umbilical cord blood: markers of storage lesion.

The continued growth in the uses of umbilical cord blood (UCB) will require the development of meaningful postthaw quality assays. This study examines both conventional and new measures for assessing UCB quality after long-term storage. The first arm of the study involved thawing UCB in storage for short (approx. 1 year) and long periods of time (>11 years). Conventional postthaw measures (colony-forming units [CFU], total nucleated cell counts, CD34+45+) were quantified in addition to apoptosis. The second arm of the study involved taking units stored in liquid nitrogen and imposing a storage lesion by storing the units in -80°C for various periods of time. After storage lesion, the units were thawed and assessed. In the first arm of the study, there was little difference in the postthaw measures between UCB stored for short and long periods of time. There was a slight increase in the percentage of CD34+45+ cells with time in storage and a reduction in the number of cells expressing apoptosis markers. When moved from liquid nitrogen to -80°C storage, the nucleated cell count varied little but there was a distinct decrease in frequency of CFUs and increase in percentage of cells expressing both early and late markers of apoptosis. Nucleated cell counts do not reflect damage to hematopoietic progenitors during long-term storage. Expression of caspases and other markers of apoptosis provide an early biomarker of damage during storage, which is consistent with other measures such as CFU and percentage of CD34+45+ cells.

Factors which can influence the quality related to cell viability of the umbilical cord blood units

Cell viability is an important indicator for the quality of umbilical cord blood (UCB) units that can influence the transplant final outcome. Thus, it is particularly important to identify the factors that may affect the cell quality during the banking process. The present study is a first attempt to correlate the impact of exogenous factors (time from collection to processing, collected UCB volume) and endogenous factors (TNCC – total nucleated cell count, CD34+cell count) on cell viability assessed before UCB units cryopreservation within a banking standardized process. Three thousand UCB units collected in 35 ml CPDA containing bags were processed by HES sedimentation within 48 h. TNCC, CD34+ cell counts and total cell viability were determined after processing. Cell viability of 94.37 ± 4.67%, TNCC of 73.17 ± 36.73 × 107 and CD34+cell count of 2.61 ± 2.29 × 106 was obtained after processing of units with UCB collected volume of 80.23 ± 28.52 ml. A significant negative correlation was found between cell viability and the time from collection to processing (r = –0.7228; P < 0.0001). The cell viability decreasing rate of 20.54%, 15.18% and 3–10% were achieved for units with collected UCB volume <40 ml, (40–80 ml) and >80 ml, to 48 h versus 12 h. There were no differences considering cell viability for the UCB units with similar collected UCB volume that had various CD34+cell count or TNCC (P > 0.05). The extension of the time from collection to processing of UCB units can reduce the quality by decreasing cell viability. The cell viability decreasing rate owing to the time influence is determined by the collected UCB volume being inversely proportional to it. Endogenous factors do not affect the cell viability.

Study for the improvement of umbilical cord blood sampling using a new trial apparatus

The aim of this study was to evaluate the usefulness of the trial umbilical cord blood sampling bag for unrelated cord blood transplantation. Data were obtained from 100 vaginal deliveries. In 50 cases, umbilical cord blood (UCB) was taken with the traditional Kawasumi type UCB sampling bag. In another 50 cases, UCB were taken with trial UCB sampling bag offered by NIPRO Co. We compared the sampling volume between the two groups. Furthermore, 10 cases in each group were matched by sampling volume; we examined the quality of UCB on the number and concentration of nucleated cells, mononuclear cells, CD34+ cells and colony-forming unit granulocyte macrophage and the numbers tested positive for bacteria. Whereas there were no significant differences in gestational weeks at sampling, the ratio of primipara women to multipara women, maternal age, and neonatal weight between the two groups, the sampling UCB volumes with the trial sampling bag were significantly higher than those with traditional sampling bags (P < 0.05). In addition, this phenomenon was more significant in the latter part of the study period (P < 0.05). On the other hand, there were no significant differences in the quality of UCB between the two groups. Once clinicians have become accustomed to the trial UBC sampling bag, this method might be a useful method for collecting UCB for unrelated cord blood transplantation.

The influences of maternal and neonatal factors on umbilical cord blood quality

The influences of maternal and neonatal factors on umbilical cord blood quality

Quality control measures in cord blood banking in India - critical appraisal and recommendations.

Umbilical cord blood (UCB) is a potential source of primitive hematopoietic stem and progenitor cells (HSPC) available for clinical application to reconstitute the hematopoietic system in patients lacking conventional stem cell donors. In the last decade because the frequency of UCB stem cell transplantation has increased, the quality of UCB available in banks has become an important criterion for the success of any UCB stem cell transplant. Moreover, with the growing numbers of cord blood banks there is not only a need to improvise quality issues but to also develop a more homogenous approach in setting standards and regulations for assessing them. The present article provides an insight on the importance of cord blood banking in India and the impending requirement to provide clarity on certain issues regarding UCB banking. Furthermore, the article discusses about certain practical approaches which could perhaps be referred to as guidelines (and by no means be solely adopted as the only measures) by cord blood banks in India for establishing or maintaining a standard for quality control (QC) measures.

Factors affecting banking quality of umbilical cord blood for transplantation

The most important objective for cord blood banks is to store cord blood units of high quality, which is determined by total nucleated cells (TNCs) and CD34+ cells. Determining the factors affecting the stored life-saving cells would be beneficial to the field. A total of 4930 cord blood units were collected between January 2007 and October 2009 and processed using a double extraction technique to sediment red blood cells with variable centrifugation time determined by the formula CT = KL - M, where CT is centrifuge time, K is 7.7227, M is 29.742, and L is ln (volume of cord blood with anticoagulant). The recovery rate of TNCs and other relevant factors affecting banking quality were analyzed. The mean recovery rate of TNCs was 97.7 ± 2.5% with 0.04% (2/4930) units below 80% and 10.8% (532/4930) units below 95%. The TNCs per unit was affected by gestation duration (p < 0.01), sex of infant (p < 0.01), mode of delivery (p < 0.01), collection method (p < 0.01), and ethnicity (p < 0.001). The number of postprocessing CD34+ cells was affected only by sex of the infant (p < 0.05). The viability of nucleated cells after processing was 94.8 ± 4.8% and was affected by the number of hours between collection and processing (p < 0.01). In contrast, the viability of CD34+ cells was 99.5 ± 1.0% (n = 30) when samples with low viability of TNCs were assessed. The results did not reveal a significant correlation (r = 0.07, p = 0.38). The double extraction technique provides a high and consistent recovery of TNCs, which ensures that more life-saving cells will be banked for transplants.

Loss of integrity of umbilical cord blood unit freezing bags: description and consequences

Umbilical cord blood (UCB) is now a commonly used resource for hematopoietic stem cell (HSC) transplantation; great effort has been put forth in standardizing protocols for processing, storage, and testing of UCB units. Because UCB units are selected on an individual basis to maximize the chance of engraftment, loss of container integrity may have adverse effects on patient outcome. All bag breaks involving UCB units thawed for transplantation at our institution between January 1, 2000, and May 31, 2006, were identified. Information on various laboratory variables and the clinical consequences of UCB bag breaks was obtained from the deviation database of the Clinical Cell Therapy Laboratory (CCTL). Patient medical charts were reviewed for infusion-related data. The incidence of bag breaks over a 6 1/2-year period was 3.5 percent. A majority of cases of loss of container integrity occurred in units that had been cryopreserved for more than 2 years (75%) and resulted in minimal loss of product. There were no significant decreases in quantity or quality of UCB, as determined by various quality control tests; no adverse clinical outcomes related to receiving a broken UCB unit were noted except increased antibiotic usage. There was a relatively low incidence of UCB bag breaks in this study that did not result in significant loss of UCB or adverse clinical outcomes. With the FDA considering licensure of UCB for hematopoietic reconstitution, improvement in container design and possibly guidelines limiting length of storage will likely be addressed in detail.

The Impact of Variability among Banks on the Quality of Umbilical Cord Blood for Transplantation.

INTRODUCTIONAs the frequency of umbilical cord blood (UCB) stem cell transplantation has increased, the quality of UCB available in banks is an important part of the success of UCB stem cell transplants.METHODSUCB units from outside banks were received in the Clinical Cell Therapy Laboratory where accompanying documentation, transfusion-transmitted disease test results, histocompatibility and ABO typing results, medical history, sterility testing, and processing records including cell counts were reviewed. Deviations were categorized based on cord blood regulations and the potential impact of deviations using a system developed to categorize according to the medical history, quality control testing, or labeling and documentation and for their potential risk to the patient.RESULTSBetween January 1, 2001 and December 31, 2003, 268 UCB units were received for transplantation from 14 UCB banks. Of these 268 units, 151 (56%) from 12 banks (86%) had a total of 246 quality issues (Table). Most (54%) of the issues were in quality control. Examples included: 21 units with transmissible diseases tests that were incomplete, pending, or with positive results; 4 with a positive microbiological screening culture at the shipping UCB bank from a sample obtained prior to cryopreservation; 20 from mothers thought to be colonized with group B streptococcus; 31 from mothers who received antibiotics during labor; and 7 units arrived with inadequate temperature monitoring so it could not be documented that the units were maintained at an acceptable temperature. 11 units had questionable quality control test results for cell viability < 50% (2), poor CFU growth (2), nucleated red cell content > 20% (5), leaking segment (1) and confirmatory HLA typing not performed from attached segment (1). Of the 132 units with quality control issues, 25 (19%) were likely to pose a risk to patients. Documentation was inadequate for five units because related records did not have a complete numbering system. Reports of transmissible disease testing, cell content, donor medical history, or other details were sent on sheets of paper that did not contain the unique number of the UCB unit or other identifying information to assure that the data on that page were from the unit in question. Very few (4%) medical history issues posed a risk to patients. Examples of the medical history issues included: maternal exposure to malaria, recent tattoo, body piercing, maternal HIV risk factors, and maternal history of herpes, chlamydia, human papilloma virus (HPV) or use of donor sperm for the pregnancy.CONCLUSIONBecause standards have evolved over time, UCB banks contain units that have different levels of quality. Some units have been placed in the usable inventory with incomplete test results and/or documentation or that may not meet the banks own current criteria. Since many are only recognized after a transplant decision has been made, information about any quality or operating procedure deviation should be provided in sufficient detail and at the initiation of the search process so that transplant physicians can consider these quality issues when making a transplant decision.TotalPotential Impact#%LikelyPotentialUnlikelyMedical history994043758Quality control13254253077Labeling/documentation1564110TOTAL2461003378135

Critical parameters for the isolation of mesenchymal stem cells from umbilical cord blood.

Evidence has emerged that mesenchymal stem cells (MSCs) represent a promising population for supporting new clinical concepts in cellular therapy. However, attempts to isolate MSCs from umbilical cord blood (UCB) of full-term deliveries have previously either failed or been characterized by a low yield. We investigated whether cells with MSC characteristics and multi-lineage differentiation potential can be cultivated from UCB of healthy newborns and whether yields might be maximized by optimal culture conditions or by defining UCB quality criteria. Using optimized isolation and culture conditions, in up to 63% of 59 low-volume UCB units, cells showing a characteristic mesenchymal morphology and immune phenotype (MSC-like cells) were isolated. These were similar to control MSCs from adult bone marrow (BM). The frequency of MSC-like cells ranged from 0 to 2.3 clones per 1 x 10(8) mononuclear cells (MNCs). The cell clones proliferated extensively with at least 20 population doublings within eight passages. In addition, osteogenic and chondrogenic differentiation demonstrated a multi-lineage capacity comparable with BM MSCs. However, in contrast to MSCs, MSC-like cells showed a reduced sensitivity to undergo adipogenic differentiation. Crucial points to isolate MSC-like cells from UCB were a time from collection to isolation of less than 15 hours, a net volume of more than 33 ml, and an MNC count of more than 1 x 10(8) MNCs. Because MSC-like cells can be isolated at high efficacy from full-term UCB donations, we regard UCB as an additional stem cell source for experimental and potentially clinical purposes.