Multilineage differentiation activity by cells isolated from umbilical cord blood: Expression of bone, fat, and neural markers

The stromal cell population in bone marrow has been the focus of much attention since it has been shown that this cell population can be expanded and differentiated into cells with the phenotype of bone, cartilage, muscle, stroma, neural, and fat cells. We evaluated umbilical cord blood (UCB) for the presence of these cells. From the mononuclear fraction of UCB, we demonstrated the presence of a subset of cells that have been maintained in continuous culture for more than 6 months (>10 passages). These adherent cell populations express adhesion molecules CD13+, CD29+, and CD44+, but not antigens of hematopoietic differentiation. Exposure of these cells to osteogenic agents resulted in an increase in expression of alkaline phosphatase and the appearance of hydroxyapatite nodules by Von Kossa staining. Incubation with adipogenic agents resulted in morphological change and staining with Oil Red O. In addition, when exposed to basic fibroblast growth factor and human epidermal growth factor the cells underwent changes consistent with cells of neural origin. These changes were demonstrated by a combination of immunofluorescent labeling and Western immunoblots for neural-specific markers. Thus, similar to what has been previously reported with bone marrow, cord blood contains a population of cells that can be expanded in culture and are able to express the phenotype of multiple lineages. Cord blood multilineage cells are slower to establish in culture, have a lower precursor frequency and a lower level of bone antigen expression, and lack constitutive expression of neural antigens when compared to bone marrow, suggesting a more primitive population. Cord blood may prove to be a new source of cells for cellular therapeutics for stromal, bone, and, potentially, neural repair. [1]

Augmentation of umbilical cord blood (UCB) transplantation with ex vivo–expanded UCB cells: results of a phase 1 trial using the AastromReplicell System

Allogeneic stem cell transplantation with umbilical cord blood (UCB) cells is limited by the cell dose a single unit provides recipients. Ex vivo expansion is one strategy to increase the number of cells available for transplantation. Aastrom Biosciences developed an automated continuous perfusion culture device for expansion of hematopoietic stem cells (HSCs). Cells are expanded in media supplemented with fetal bovine serum, horse serum, PIXY321, flt-3 ligand, and erythropoietin. We performed a phase 1 trial augmenting conventional UCB transplants with ex vivo–expanded cells. The 28 patients were enrolled on the trial between October 8, 1997 and September 30, 1998. UCB cells were expanded in the device, then administered as a boost to the conventional graft on posttransplantation day 12. While expansion of total cells and colony-forming units (CFUs) occurred in all cases, the magnitude of expansion varied considerably. The median fold increase was 2.4 (range, 1.0-8.5) in nucleated cells, 82 (range, 4.6-266.4) in CFU granulocyte-macrophages, and 0.5 (range, 0.09-2.45) in CD34+ lineage negative (lin–) cells. CD3+ cells did not expand under these conditions. Clinical-scale ex vivo expansion of UCB is feasible, and the administration of ex vivo–expanded cells is well tolerated. Augmentation of UCB transplants with ex vivo–expanded cells did not alter the time to myeloid, erythroid, or platelet engraftment in 21 evaluable patients. Recipients of ex vivo–expanded cells continue to have durable engraftment with a median follow-up of 47 months (range, 41-51 months). A randomized phase 2 study will determine whether augmenting UCB transplants with ex vivo–expanded UCB cells is beneficial. [2]

Collecting and analyzing cord blood gases

The analysis of cord blood respiratory gases and acid-base values is an important adjunct for determining the extent and cause of fetal acidosis at delivery. Although the quality and reliability of the blood gas instruments have improved dramatically, constant vigilance still is required and mandated to ensure accurate and precise results. Failure to control the many sampling and analysis variables that affect cord blood gas results will limit their usefulness. Most preanalytic problems become a minor concern when the blood gas analyses are done within a few minutes after obtaining the sample. Comparison of data among centers requires, not only that reference ranges be stated, but also that various corrections or factors that were used to adjust the results be described. Perhaps, a consensus could be reached to establish the optimal method of collection and the best methods for analyzing and reporting the results from cord blood gas and acid-base studies. [3]

Distribution of Telomere Length in the Cord Blood of Chinese Newborns

Aims: We studied the variability in telomere length in cord blood collected from newborns of different birth weights and gestational ages.

Study Design: Prospective cohort study.

Place and Duration of Study: Samples were collected from KK Women’s and Children’s Hospital between March 2011 and March 2012 and the terminal restriction fragment assays (TRF) were performed at the Department of Physiology, National University of Singapore.

Methodology: Cord blood samples were prospectively collected in EDTA or heparin tubes for deliveries from Chinese parents. TRF assays were performed on genomic DNA extracted from whole blood. Data was collected for birth weight, gestational age, and maternal age. Variance analyses of telomere lengths and correlation coefficients were calculated using Statistical Package for the Social Sciences (SPSS).

Results: The birth weight of the samples collected ranged from 0.61 kg to 5.18 kg with gestation age from 196 to 288 days. TRF results from 184 samples (96 males, 88 females) showed that there was a wide range from 6.6 kbp to 19.2 kbp. The mean TRF length was 12.64 kb (males: 12.33 kb ± 2.50; females: 12.99 kb ± 2.35). There was no statistically significant correlation of TRF with birth weight, gestation age or maternal age. There was highly significant correlation of birth weight with gestational age (P=0.00).

Conclusion: Our results showed no correlation of either gestational age or birth weight with telomere length as measured by TRF assay. [4]

Utility of Placental Umbilical Cord Blood in Autoimmune and Degenerative Disorders

Background: Umbilical cord blood is whole human blood (60 to 80 ml) that remains in the placenta and umbilical cord after childbirth; generally considered as a medical waste. It is a rich source of stem cells, growth factor, cytokines, etc., and, can be collected, stored and utilized in the treatment of incurable diseases.

Aims and Objects: The aim of the present study is to establish the fact that placental umbilical cord whole blood is a safe alternative to adult blood and to assess its utility in degenerative and autoimmune disease along with its hematological parameters.

Materials and Methods: It is a prospective two year study (From September 2016 to August 2018) of 250 umbilical cord whole blood transfusions in autoimmune and degenerative disorders at Gajra Raja Medical College, Gwalior, India. Follow up of patients was done up to 3 months and data was collected and analyzed statistically by frequency distribution and percentage proportion.

Results: A total of 250 units meeting the inclusion and exclusion criteria were transfused to 99 preregistered patients; Vitiligo 61 (159 transfusions), Thalassemia 15 (30), Retinitis Pigmentosa 9 (23), Geriatric Disorders 9 (24), Aplastic anemia 4 (9) and High Myopia 1 (5). Out of 250 transfusions, in one case (0.4%) adverse event was reported. Outcome of transfusion reveals; In Vitiligo –regimentation in affected area, Thalassemia-reduction in frequency of transfusions, Retinitis Pigmentosa- improvement in vision area, Geriatric patients- sense of well being, Aplastic anemia- prolonged survival and High Myopia-improvement in vision area.

Conclusion: Umbilical cord blood is safe and genuine alternative of adult blood. It is effective in degenerative and autoimmune diseases. It should not be discarded as medical waste and utilized judiciously in the human well being. [5]

Reference

[1] Goodwin, H.S., Bicknese, A.R., Chien, S.N., Bogucki, B.D., Oliver, D.A., Quinn, C.O. and Wall, D.A., 2001. Multilineage differentiation activity by cells isolated from umbilical cord blood: expression of bone, fat, and neural markers. Biology of Blood and Marrow Transplantation, 7(11), pp.581-588.

[2] Jaroscak, J., Goltry, K., Smith, A., Waters-Pick, B., Martin, P.L., Driscoll, T.A., Howrey, R., Chao, N., Douville, J., Burhop, S. and Fu, P., 2003. Augmentation of umbilical cord blood (UCB) transplantation with ex vivo–expanded UCB cells: results of a phase 1 trial using the AastromReplicell System. Blood, 101(12), pp.5061-5067.

[3] Riley, R.J. and Johnson, J.W., 1993. Collecting and analyzing cord blood gases. Clinical obstetrics and gynecology, 36(1), pp.13-23.

[4] Lim, S.-N., Yahya, Z., Zeegers, D., Kyaw, E. E., Yeo, G., Hande, M. and Tan, E.-C. (2013) “Distribution of Telomere Length in the Cord Blood of Chinese Newborns”, Journal of Advances in Medicine and Medical Research, 3(4), pp. 1004-1014. doi: 10.9734/BJMMR/2013/2676.

[5] Khatoon, M., Bindal, J., Chandra Sharma, D., Gupta, P., Singh Tomar, A., Saify, K. and Gupta, R. (2018) “Utility of Placental Umbilical Cord Blood in Autoimmune and Degenerative Disorders”, International Blood Research & Reviews, 8(4), pp. 1-9. doi: 10.9734/IBRR/2018/45260.