Maternal and Neonatal Factors' Effects on Wharton's Jelly Mesenchymal Stem Cell Yield

doi:10.21203/rs.3.rs-4437032/v1

Download PDF

Article

Maternal and Neonatal Factors' Effects on Wharton's Jelly Mesenchymal Stem Cell Yield

https://doi.org/10.21203/rs.3.rs-4437032/v1

This work is licensed under a CC BY 4.0 License

You are reading this latest preprint version

Because Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) are easily accessible, easy to isolate, and ethically acceptable, they represent a talented source of MSCs for use in regenerative medicine. Considering decisions on WJ-MSCs collection requires extensive knowledge of the factors that impact their yield. This study's objective was to evaluate the influence of parameters related to mothers and newborns on the WJ-MSCs yield. The WJ-MSCs were isolated and expanded after being taken from 79 umbilical cord samples. Population doubling time and cell proliferation were assessed. By flow cytometry analysis, WJ-MSCs were identified by positivity of CD105, CD90, and CD73 and negativity of CD45 and CD34. There was a statistically significant negative correlation between umbilical cord width and P1 doubling time. Maternal age and WJ-MSC yield were shown to be negatively correlated. Birth weight and gestational age showed a significant positive correlation between WJ-MSCs yield and neonatal variables. No significant correlations were detected between the WJ-MSCs and the mother parity, nor the neonatal sex, fetal presentation, or head circumference. The WJ-MSCs yield increases with younger maternal age, increased gestational age, and increased neonatal birth weight. Hence, consideration should be given to these factors when selecting the ideal donors.

Biological sciences/Stem cells

Health sciences/Medical research

Wharton's jelly

Stem cells

Yield

Maternal- Neonatal

Flow cytometry

The stem cells are a special type of cells that have a potentiality for regeneration and thus can maintain and repair cellular damage. So, it have a promising therapeutic prospective in cell transplantation. Mesenchymal stem cells (MSCs), hematopoietic stem cells, and other progenitors are among the cells from different origins that have been isolated Anthony and Shiels [1] [2]. Mesenchymal stem cells (MSCs) isolated from bone marrow have been the focus of most studies; however, MSCs may also be obtained from amniotic fluid and umbilical cord (UC) blood [3] and chorionic villi [4]. The mesenchymal stem cells have been also derived from the Wharton's jelly, the connective tissue that envelops the two arteries and vein in the UC [5].

WJ-MSCs have grown increasingly common in recent years due to their inherent advantages than MSCs obtained from other sources (Baksh et al., 2007). These advantages include increased ex-vivo growth capabilities, faster proliferation, a lower incidence of graft versus host medical problems, and a lower risk of teratomas [6]. Additionally, there are no ethical issues involved in obtaining them, in contrast to bone marrow and embryonic stem cells, which are often thrown as human waste [7]. Wharton's jelly mesenchymal stem cells (WJ-MSCs) possess paracrine activity, which is the ability to alter their environment by releasing bioactive chemicals and compounds known as secretome. This activity is the primary reason for the therapeutic potential of WJ-MSCs. The secretome has anti-inflammatory and anti-fibrotic properties, but it also carries out several biological activities like immuno-modulation, tissue replenishment, and cellular homeostasis [8]. Previous researches recommended its use for cancer cells growth inhibition [9], ameliorating diabetic nephropathy and hepatopathy complications as a novel protective approach [10], the reduction of the severity of the inflammation and immune response against Covid-19 because of its anti-inflammatory and immunomodulatory characteristics [11], and reduce fibrosis caused by bleomycin-induced lung injury [12].

There are many approaches to identifying mesenchymal stem cells as their capacity to adhere to flexible surfaces and their spindle-like appearance in culture and they can differentiated into adipocytes, chondrocytes and osteocytes. Furthermore, WJ-MSCs should express CD105, CD73 and CD90 and not display CD34, CD45, CD19 or CD11b, CD79 alpha or CD14, or HLA-DR surface molecules, according to the International Society for Cellular Therapy (ISCT) [13]. Since isolating cells mostly implies an invasive and technically challenging procedure that affects the quality and biological activity of the derived cells and restricts their ability to proliferate and differentiate during in vitro culture, the quantity of retrieved stem cells is inadequate [14]. It is essential to gain a good yield from the WJ-MSCs. Many factors have been suggested to impact the quantity of WJ-MSCs isolated from Wharton’s jelly, and which may be accountable for the variations in the reported results. These variables include the mother's age, the gestational age, mother’s parity and gravidity, the newborn's sex, and the birth weight, all of which might affect the amount of CD34 + cells in the blood [15].

Considering the maternal and neonatal factors in the selection of promising donors for isolation of a good yield from WJ-MSCs, our study aimed to determine whether maternal parameters-such as the mother's age, gravidity, parity, and other health-related conditions—and neonatal parameters—such as birth weight, sex, head circumference, and UC width—may affect the yield of WJ-MSCs.

The Institutional Review Board Mansoura Faculty of Medicine, Mansoura University, Egypt approved the current study, and informed consents were obtained from all participants before delivery. A total of 100 pregnant women were included in the study. Women with temperature higher than 38^oC, having premature rupture of membranes, with multiple gestation, positive for blood transmitted infections or with proved fetal malformations were excluded from the study. The patient records were reviewed for information on pregnancy history, number of prior pregnancies, parities, the age of gestation, newborn sex, newborn weight, and head circumference, the width of UC, gestational diabetes, pre-eclampsia, and any reports of maternal co-morbidity. Patients' records and gathered data for the study were kept confidential and experiments were performed in accordance with relevant guidelines and regulations.

Samples collection and cord processing:

Following cesarean birth, a total of 100 fresh human UC samples were collected. A collection cup holding in Dullbecco's Modified Eagle Media (DMEM) with 4,500 mg/mL glucose and antibiotic solution (.2% streptomycin,, .12% penicillin and .1% gentamicin) (Lonza, Belgium) was used to hold the roughly 5 gm of UC that were aseptically collected. The samples were subsequently kept at 4°C. After that, the samples were sent to the Mansoura University Stem Cell Research Center lab to be processed. In order to eliminate blood clots, samples were processed in a biosafety cabinet and washed using ice-cold phosphate buffered saline repeatedly.

The explant method was utilized to isolate mesenchymal stem cells from the UC Wharton's jelly [16]. Six to nine pieces of the explant outgrowth are cut off and put on the culture dishes, where they remain until the jelly hardens. Next, let's include a culture media (DMEM). The culture dishes are kept for 3 to 4 days at 37°C in a humidified atmosphere with 5% CO2. Following the attachment of Wharton jelly, the dish media is replaced every 2 to 3 days, and after about 7 to 10 days, the cells displayed the WJ-MSCs phenotype will be recovered. The cells were separated by the use of 1–2 mL of commercial trypsin solution in a 25-cm² culture flask before being incubated for 3 min. at 37°C. Using a tissue culture centrifuge, the mixture was centrifuged at 1,500×g for three minutes in order to neutralize the trypsin. One to two milliliters of culture medium were added. To carry out amplification and characterization, the pelleted cells—which are regarded as passage 0 (P0)—were re-suspended in a culture medium, counted, and sub-cultured at a 1x10⁴/cm² seeding density.

Cell viability determination and Population Doubling Time determination:

The cell viability was determined using the Trypan blue test as described by [17]. After the isolated cells were stained with Trypan blue, the number of viable (white) and non-viable (blue) cells was determined using a hemocytometer composed of nine 1×1 mm (1mm²) squares and a microscope (AX-71, Olympus Corporation, Shinjuku-ku, Japan). Every experiment was carried out three times in duplicate.

Trypsin was used to harvest the cells at 80–90% confluence after each passage. The cells were then plated in a T25 cell culture flask, counted, and re-plated till the third passage. The population doubling time (PDT) was calculated according to [18]. The following formula was used for calculating PDT, which was expressed in hours: PDT is equal to (lgNt – lgN0)/lg2, where t is the culture period, Nt is the harvested cell count after the cell passage, and N0 is the number of cells seeded at the start of the passage.

Flow cytometry:

Using the International Society for Hematotherapy and Graft Engineering (ISHAGE) procedure [19], flow cytometric analysis was used to determine if the isolated cells were multipotent WJ-MSCs and to evaluate the WJ-MSCs yield. The BD Accuri TM C6 Cytometer (Becton, Dickinson and Company) was used. The program System (Becton, Dickinson and Company) was used for analyzing the data [20]. According to [21], the WJ-MSCs were characterized by positivity of CD73, CD90, CD105 and negativity of CD34 and CD45 on flow cytometry analysis as shown in figure (1).

In brief, after being re-suspended in 1 ml of PBS including 1% FBS, the cultured WJ-MSCs were labeled with 5µl of fluorescein isothiocyanate (FITC), APC anti-human, and phycoerythrin (PE)-conjugated antibodies as positive markers for CD105, CD90, and CD73 respectively. Antibodies conjugated with phycoerythrin cyanine 5 (PE-Cy5) and PE-Cy7 were utilized as negative markers for CD45 and CD34, respectively. As controls, FITC, PE, and PC5 matched isotype antibodies were used. All antibodies were from BD Biosciences except CD90,and CD73 were from Miltenyil Biotec. At 4°C and in the dark for 45 minutes, antibodies were added at 1:1000 dilutions then the yield was assessed using flow cytometry.

Statistical analysis:

The collected data was revised, coded, tabulated and introduced to Statistical package for Social Science (IBM SPSS Statistics, Version 25.0. Armonk, NY). Mann Whitney-U or student t-test was applied for comparing numerical variables between groups. Chi-square test was utilized to compare categorical variables. Correlation analysis was used for studying the strength of correlation between two numerical variables. The correlation between WJ-MSCs yield at the end of P3 and P1 doubling time and the potential numerical maternal and newborn predictors was tested by Spearman correlation coefficient. A correlation coefficient of less than .3, between .3-.7 and of more than .7 indicated weak, moderate and strong linear correlations, respectively.

Demographics:

At the Mansoura University Hospital delivery unit, UC samples were obtained from 100 singleton newborns who were delivered via Caesarean section. Twenty-one specimens were excluded because, in 14 samples, the material was contaminated, and in 7 samples, there was no apparent cellular growth after 20 days. The final analysis had 79 specimens in total. The maternal data were as follows: the mean parity was 1.2 ± 1.1, the mean gravidity was 2.4 ± 1.3, and the mean age was 29.3 ± 5.4 years, with a range of 18 to 42 years. Four (5.1%) of the mothers in the study have gestational diabetes; two were managed with diet, and the other four required insulin therapy. There were six mothers (7.6%) with pre-eclampsia. Of the seventy-nine babies who were part of the study, fifty-one (64.6%) were female and twenty-eight (35.4%) were male. Of the infants born between 26 and 41 weeks, 56 (70.9%) were full term and 23 (29.5%) were preterm. The mean gestational age was (36.3 ± 3.3). The median birth weight was 2.8 (1.3–4) kg, with a mean of 2.73 ± .6 kg. The median birth length was 45 (39–52) cm, with a mean of 45.5 ± 2.99 cm. Within the 30–35 range, the average newborn head circumference was 33.1 ± 4.8. Ten (12.7%) newborns had amniotic fluid stained with meconium, while sixty (75.9%) babies were cephalic presentation.

Effects of neonatal and maternal variables on the passage (P1) doubling time:

For WJ-MSCs, the average P1 doubling time was 68 (55–90) hours. When the association between P1 doubling time and maternal variables were analyzed, Fig. 2A, 2B, and 2C showed that P1 doubling time had a small positive correlation with gravidity (r = .624, P < .001), parity (r = .551, P < .001), and a strong positive correlation with maternal age (r = .827, P < .001). P1 doubling time was not significantly associated with any of the neonatal variables: birth head circumference (P = .543), birth length (P = .236), birth weight (P = .735), or gestational age (P = .186). There was no statistically significant difference in the median doubling time between preterm newborns (69 hours) and full term babies (67.5 hours) (P = .612). In addition, there was no statistically significant difference in the hours of male (66) and female (68) babies (P = .612). The presence of amniotic fluid stained with meconium (P = .238) or fetal mal-presentation (P = .502) had no discernible effect on the P1 doubling time. Double time and UC width were shown to have a statistically significant negative connection (r=-.5, P = .001) (Fig. 2D).

Maternal and neonatal variables' effects on the yield of WJ-MSCs:

By the end of P3, the mean WJ-MSCs yield was (2.34 ± .74×106) cell/ml [2.6 (.4–3.5) ×106 cell/ml, according to the median (range). For the median (range), the viability percentage was 83.1 ± 7.6% [84.5 (65–97)%]. UC samples were used to determine the WJ-MSCs count of full-term neonates (2.37 ± .74 x 106 cell/ml; median = 2.6 (.4–3.5)×106 cell/ml) and preterm newborns (2.27 ± .76 x 106 cell/ml; median = 2.6 (.7–3.3)×106 cell/ml). No statistically significant difference was seen in the median WJ-MSCs yield between preterm and full-term newborns (Mann-Whitney Up = .742).

Maternal age and WJ-MSCs yield had a negative correlation (r=-.246, P = .029) according to an analysis of maternal variables (Table 1). There was no statistically significant difference in the median WJ-MSCs yield between UC specimens obtained from women with and without gestational diabetes. In UC specimens collected from mothers with or without pre-eclampsia, WJ-MSCs yield was also similar. It is noteworthy, therefore, that there were very few pre-eclamptic or gestational diabetic individuals in the study. WJ-MSCs yield did not significantly correlate with maternal hemoglobin level prior to birth, parity, or gravidity.

Analyses of the association between WJ-MSCs yield and newborn characteristics (Table 1) revealed a positive correlation between the WJ-MSCs count and birth weight and gestational age. However, it was shown that the yield of WJ-MSCs and UC width had a weakly positive correlation. A multivariate linear regression analysis revealed that the sole independent predictor of WJ-MSCs production was maternal age (Table 2). The yield of WJ-MSCs was not independently predicted by gestational age, birth weight, or cord breadth (P = 0.330).

A promising source of MSCs for therapeutic uses is Wharton's Jelly. WJ-MSCs are easily accessible with many advantages over the BM-MSCs. Several maternal and neonatal parameters have been studied in relation to the characterization and quality of hematopoietic UC blood stem cells in the past. Although research on how maternal and neonatal variables affect the yield of WJ-MSCs is still under research investigation. For the purpose of aiding in finding and selecting the best UC donors, this study studied the effects of many maternal and fetal factors on the yield of WJ-MSCs.

According to our findings, the mother's age has a substantial impact on the yield of WJ-MSCs. This is in agreement with the findings of [22], who found that whereas UC-MSCs from elder donors show diminished differentiation potential, UC from younger donors is a reasonably abundant source of MSCs. Furthermore, compared to younger age groups, there was a significant negative correlation in the expression levels of both CD29 and CD105, according to [23]. It was reported that as a woman ages, her mesenchymal stem cells expressing the BIRC2 and BIRC3 genes decrease as stem cells obtained from younger women may exhibit more apoptotic resistance and a more stem cell-like quality, which may enhance their therapeutic potential and clinical usability [16].

A known pluripotent transcription factor, SOX2 is implicated in reprogramming, self-renewal, and maintaining the homeostasis of stem cells. The SOX2 gene is statistically considerably more expressed in WJSC in women under the age of 34 than it is in women over 34, according to [24]. Furthermore, a statistically significant moderate negative correlation is demonstrated between the SOX2 gene expression and the maternal age. This result was clarified by [25] and [26], who found that aging in mothers is related to telomere shortening and increased susceptibility to apoptosis, and that stem cells' telomere length could not be maintained by the level of telomerase activity.

These results largely stated the decline in CD105, CD73, and CD90 expression levels that they reported in their study's UC-MSCs as they were older. Furthermore, it has been previously reported how aging affects adult stem cells. [27] found a decrease in adult MSCs number derived from the bone marrow with aging. According to [28], there is a decrease in the quantity, functionality, and multi-lineage differentiation characteristics of mature MSCs as they age. Additionally, the authors hypothesized that the lower differentiation capacity of the older donor's umbilical cord cells could be related to the declining functional state of the mother's older organs, which support and foster the development of umbilical cord MSC [22]. Conversely to this finding, maternal age had no effect on the quality of WJ-MSCs, according to [29].

According to [30], vaginal and cesarean births had MSC cell yields that were comparable. According to [29], the method of delivery had no impact on the characteristics or survivability of MSCs. To reduce the expectation of contamination, we only used specimens from Cesarean births in our investigation. We observed that the WJ-MSCs count and doubling time were not more impacted by birth order. A reduction in the number of WJ-MSCs was correlated with increasing parity, nevertheless the relationship was not statistically significant. Birth order and viability showed a moderately positive association (r = .119, p = .044), whereas parity and the amount of UC blood showed no correlation (P = .057) [31]. According to [32], WJ-MSCs obtained from younger women who gave vaginal delivery have increased expression of the BIRC2, BIRC3, and BIRC5 genes.

Pre-eclampsia had no obvious effect on the population doubling time or WJ-MSC yield in this study. In contrast, pre-eclampsia has been shown by [33] to result in a considerable increase in cell count and reduction in the fraction of immature cells and increasing the mature cells. According to [34] compared to normal samples, the proliferation rate of early-passage gestational diabetes mellitus (DM) samples was significantly lower. Furthermore, with a mean cell density of 1.9 x 10⁴ cells per plate in gestational DM samples as opposed to 4.1×10⁴ cells per plate in normal samples, the population doubling rate was considerably lower in these samples after 12 days of incubation. Gestational DM-UC-MSCs did not grow above passages 6 or 7. All UC-MSCs had comparable amounts of the stem cell markers CD90, CD73, and CD105 during early stages. These results are consistent with a study conducted by [35], which found that MSCs derived from Wharton Jelly of the DM group grew and proliferated at a much slower rate in all passages than MSCs isolated from a control group. The quantity of non-viable cells in the gestational DM samples significantly increased than viable cells.

The WJ-MSCs count was shown to be statistically significantly impacted by gestational age in our investigation, although there was not a significant association established between gestational age and P1 doubling time. Nevertheless, there was no statistically significant difference in the median MSC output between full-term and preterm newborn when comparing them to full-term babies. In a study by [36] divided 45 babies into three groups based on their gestational ages: group A (28–31 weeks), group B (32–35 weeks), and group (C) ≥ 36 weeks. The authors observed a significant inverse relationship between gestational age and MSC production, even after controlling for birth weight, the percentage of small for gestational age neonates, maternal co-morbidities, and prenatal steroid usage. Similar findings were made by [37], who found that despite both samples maintained a high rate of proliferation, preterm cord samples developed more MSC in all passages than term cords. On the other hand, according to [38], the quantity of umbilical cord vein UC-MSCs from both full term and preterm patients is comparable, and both groups exhibit a high rate of proliferation. [29], on the other hand, found no correlation between gestational age and MSC yield.

Age at gestation may possibly have an impact on the UC-MSCs' potential to modulate immunity. According to [38], preterm UC-MSCs do not have the same potent inhibitory effects on immune cell proliferation as full-term UC-MSCs. A lymphocyte co-culture experiment revealed that both full term and preterm UC-MSCs reduced peripheral blood mononuclear cell proliferation in the mixed lymphocyte response. Compared to preterm UC-MSCs, the term UC-MSCs had a stronger proliferation inhibition. The scientists explained the inhibitory difference by the preterm UC-MSCs' immature immune systems. The source, donor age, and passage number of cells can also have an impact on the immuno-modulatory qualities of MSCs [39] and [40].

According to [41], placental weight, birth weight, and full term birth were the primary factors influencing cell proliferation. Despite the strong correlation between these parameters and gestational age, multivariate regression analysis revealed that gestational age was the only independent predictor of population doubling time (P = .0094). Our study found that the baby's birth weight had a substantial impact on the number of WJ-MSCs; a greater birth weight leads to increased the yield of MSCs. According to [30], the yield of MSCs was not significantly affected by the mother's age or gestational age, but a greater birth weight was associated with a more yield, which is consistent with our findings. On the other hand,[24] found that lower birth weight correlates to higher SOX2 gene expression in WJ-SCs. SOX2 is a known pluripotent transcription factor that contributes to cell reprogramming and self-renewal. According to [29], the MSCs output was not significantly impacted by birth weight. The author explained that his study focused on the quality of the stem cells not the number so the study finding differs from other studies that correlate the number of isolated stem cells to the birth weight [31] [42].

According to our findings, there was no statistically significant variation in the WJ-MSCs yields with respect to fetal gender, presentation, length at birth, or head circumference. According to [36], the mean MSCs number of female neonates was greater than that of male infants. When comparing MSCs separated from WJ from female individuals to MSCs obtained from WJ from male subjects, [43] observed that the expression of OCT4 and DNMT1 genes was considerably greater in WJ-MSCs derived from male participants. They came to the conclusion that gender significantly affects stemness genes. However, [29] found that gender did not affect the quality of WJ-MSCs.

We discovered a statistically significant negative association between doubling time and UC width and a slight positive correlation between MSCs yield and UC width.

We discovered that the amount of meconium in the amniotic fluid had no effect on the production of MSCs or the population doubling time. According to [44], there was little change in the expression of the mesenchymal markers CD90 and CD105 following exposure to meconium. Amniotic fluid MSCs may change in phenotype after being exposed to meconium, although they are still capable of differentiating.

To sum up, Wharton's jelly MSCs are a perfect and exciting source of MSCs. Numerous factors related to mothers and newborns may impact the MSC output and should be investigated for settling the optimal selection criteria for the ideal donors. Our research showed that a higher yield of MSCs produced from Wharton's jelly correlates to a younger mother's age, increased birth weight, and longer gestational age.

Competing Interests:

The authors declare no competing interests.

Author Contribution

B.M., B.A. and Y.S. designed and supervised the work; M.R, D.A. and Y.S. performed the experiment. B.M., B.A. and M.R. performed the analysis; D.A., E.N, M.R. and Y.S. drafted the manuscript and designed the figures. B.M., B.A. and Y.S. aided in interpreting the results and worked on the manuscript. All authors discussed the results and commented on the manuscript.

Data Availability

"The data that support the findings of this study are available on request from the corresponding author, E.N."

Anthony, D.F. and P.G. Shiels, Exploiting paracrine mechanisms of tissue regeneration to repair damaged organs. Transplantation research, 2013. 2: p. 1–8.
Zimran, E., et al., Expansion and preservation of the functional activity of adult hematopoietic stem cells cultured ex vivo with a histone deacetylase inhibitor. Stem cells translational medicine, 2020. 9(4): p. 531–542.
int Anker, P.S., et al., Amniotic fluid as a novel source of mesenchymal stem cells for therapeutic transplantation. Blood, 2003. 102(4): p. 1548–1549.
Igura, K., et al., Isolation and characterization of mesenchymal progenitor cells from chorionic villi of human placenta. Cytotherapy, 2004. 6(6): p. 543–553.
Wang, H.S., et al., Mesenchymal stem cells in the Wharton's jelly of the human umbilical cord. Stem cells, 2004. 22(7): p. 1330–1337.
Fong, C.-Y., et al., Human Wharton’s jelly stem cells have unique transcriptome profiles compared to human embryonic stem cells and other mesenchymal stem cells. Stem Cell Reviews and Reports, 2011. 7: p. 1–16.
Lange-Consiglio, A., et al., Isolation, molecular characterization, and in vitro differentiation of bovine Wharton jelly–derived multipotent mesenchymal cells. Theriogenology, 2017. 89: p. 338–347.
Drobiova, H., et al., Wharton’s jelly mesenchymal stem cells: a concise review of their secretome and prospective clinical applications. Frontiers in Cell and Developmental Biology, 2023. 11.
Süleymanoğlu, M., et al., In vitro anti-leukemic effect of Wharton’s jelly derived mesenchymal stem cells. Molecular Biology Reports, 2024. 51(1): p. 595.
Karimi, Z., G. Daryabor, and F. Masjedi, Effects of conditioned media derived from human Wharton’s jelly mesenchymal stem cells on diabetic nephropathy and hepatopathy via modulating TGF-β and apelin signaling pathways in male rats. BMC Endocrine Disorders, 2024. 24(1): p. 6.
Gupta, G., et al., Hope on the horizon: Wharton's jelly mesenchymal stem cells in the fight against COVID-19. 2023, Future Medicine. p. 675–678.
Moodley, Y., et al., Human umbilical cord mesenchymal stem cells reduce fibrosis of bleomycin-induced lung injury. The American journal of pathology, 2009. 175(1): p. 303–313.
Horwitz, E.M., et al., Clarification of the nomenclature for MSC: The International Society for Cellular Therapy position statement. Cytotherapy, 2005. 7(5): p. 393–395.
Liau, L., et al., Characteristics and clinical applications of Wharton’s jelly-derived mesenchymal stromal cells. Current research in translational medicine, 2020. 68(1): p. 5–16.
Nakagawa, R., et al., Analysis of maternal and neonatal factors that influence the nucleated and CD34 + cell yield for cord blood banking. Transfusion, 2004. 44(2): p. 262–267.
Gil-Kulik, P., et al., Potential involvement of BIRC5 in maintaining pluripotency and cell differentiation of human stem cells. Oxidative medicine and cellular longevity, 2019. 2019.
Avelar-Freitas, B., et al., Trypan blue exclusion assay by flow cytometry. Brazilian journal of medical and biological research, 2014. 47: p. 307–315.
Miyazawa, K., et al., Characterization of newly established bovine intestinal epithelial cell line. Histochemistry and cell biology, 2010. 133: p. 125–134.
Sutherland, D.R., et al., The ISHAGE guidelines for CD34 + cell determination by flow cytometry. Journal of hematotherapy, 1996. 5(3): p. 213–226.
Mennan, C., et al., Isolation and characterisation of mesenchymal stem cells from different regions of the human umbilical cord. BioMed research international, 2013. 2013.
Dominici, M., et al., Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 2006. 8(4): p. 315–317.
Huang, S., et al., Dissimilar characteristics of umbilical cord mesenchymal stem cells from donors of different ages. Cell and tissue banking, 2013. 14: p. 707–713.
Alrefaei, G.I., et al., Effects of maternal age on the expression of mesenchymal stem cell markers in the components of human umbilical cord. Folia Histochemica et Cytobiologica, 2015. 53(3): p. 259–271.
Gil-Kulik, P., et al., Evaluation of the impact of pregnancy-associated factors on the quality of Wharton’s Jelly-derived stem cells using SOX2 gene expression as a marker. International Journal of Molecular Sciences, 2022. 23(14): p. 7630.
Mimeault, M. and S.K. Batra, Great promise of tissue-resident adult stem/progenitor cells in transplantation and cancer therapies. Stem Cell Transplantation, 2012: p. 171–186.
Guerrero, E.N., et al., Increased proliferation and differentiation capacity of placenta-derived mesenchymal stem cells from women of median maternal age correlates with telomere shortening. Aging (Albany NY), 2021. 13(22): p. 24542.
Levi, B. and S. Morrison. Stem cells use distinct self-renewal programs at different ages. in Cold Spring Harbor symposia on quantitative biology. 2008. Cold Spring Harbor Laboratory Press.
Yu, K.-R. and K.-S. Kang, Aging-related genes in mesenchymal stem cells: a mini-review. Gerontology, 2013. 59(6): p. 557–563.
Penolazzi, L., et al., Influence of obstetric factors on osteogenic potential of umbilical cord-derived mesenchymal stem cells. Reproductive Biology and Endocrinology, 2009. 7: p. 1–7.
Rezniczek, G.A., et al., Factors influencing yield and neuronal differentiation of mesenchymal stem cells from umbilical cord blood and matrix. Regenerative Medicine, 2016. 11(5): p. 465–474.
Al-Deghaither, S.Y., Impact of maternal and neonatal factors on parameters of hematopoietic potential in umbilical cord blood. Saudi medical journal, 2015. 36(6): p. 704.
Gil-Kulik, P., et al., Increased expression of BIRC2, BIRC3, and BIRC5 from the IAP family in mesenchymal stem cells of the umbilical cord wharton’s jelly (WJSC) in younger women giving birth naturally. Oxidative medicine and cellular longevity, 2020. 2020: p. 1–12.
Cervera, A., et al., Flow cytometric assessment of hematopoietic cell subsets in cryopreserved preterm and term cord blood, influence of obstetrical parameters, and availability for transplantation. American journal of hematology, 2006. 81(6): p. 397–410.
Kim, J., et al., Umbilical cord mesenchymal stromal cells affected by gestational diabetes mellitus display premature aging and mitochondrial dysfunction. Stem Cells and Development, 2015. 24(5): p. 575–586.
Wajid, N., et al., The effect of gestational diabetes on proliferation capacity and viability of human umbilical cord-derived stromal cells. Cell and tissue banking, 2015. 16: p. 389–397.
Jain, A., et al., Does mesenchymal stem cell population in umbilical cord blood vary at different gestational periods? The Indian Journal of Pediatrics, 2013. 80: p. 375–379.
Ravera, S., et al., Mesenchymal stem cells from preterm to term newborns undergo a significant switch from anaerobic glycolysis to the oxidative phosphorylation. Cellular and molecular life sciences, 2018. 75: p. 889–903.
Abolhasani, M., et al., Immunomodulatory properties of umbilical cord vein mesenchymal stromal cells influenced by gestational age and in vitro expansion. Immunology Letters, 2018. 194: p. 62–68.
Zhang, X., et al., Therapeutic effect of human umbilical cord mesenchymal stem cells on neonatal rat hypoxic–ischemic encephalopathy. Journal of Neuroscience Research, 2014. 92(1): p. 35–45.
Wang, Q., et al., Comparative analysis of human mesenchymal stem cells from fetal-bone marrow, adipose tissue, and Warton's jelly as sources of cell immunomodulatory therapy. Human Vaccines & Immunotherapeutics, 2016. 12(1): p. 85–96.
Avercenc-Léger, L., et al., Umbilical cord-derived mesenchymal stromal cells: predictive obstetric factors for cell proliferation and chondrogenic differentiation. Stem cell research & therapy, 2017. 8: p. 1–13.
Mazzoccoli, G., et al., Time related variations in stem cell harvesting of umbilical cord blood. Scientific Reports, 2016. 6(1): p. 21404.
Balzano, F., et al., Lessons from human umbilical cord: Gender differences in stem cells from Wharton’s jelly. European Journal of Obstetrics & Gynecology and Reproductive Biology, 2019. 234: p. 143–148.
Jensen, T., J. Shui, and C. Finck, The effect of meconium exposure on the expression and differentiation of amniotic fluid mesenchymal stem cells. Journal of Neonatal-Perinatal Medicine, 2017. 10(3): p. 313–323.

Table (1): A summary of the correlation between neonatal and maternal variables and the yield of WJ-MSCs

Maternal variables	Correlation coefficient	P value
Mother's age (years)	-.246*	.029*
Mother's hemoglobin level (gm/dl)	.037	.743
Parity	.069	.545
Gravidity	.003	.982
Toxemia		.732**
Gestational diabetes		.762**
Neonatal variables
Gestational age in weeks	.329	.004*
Sex		.681**
Fetal presentation		.373**
Newborn birth weight (kg)	.272	.015*
The width of Umbilical Cord (cm)	.285	.011*
Birth length (cm)	.057	.620
The head circumference (cm)	.130	.253
Meconium stained amniotic fluid		.308**

P values < .05*, **Mann-Whitney U test.

Table (2): The WJ-MSCs yield model using linear regression analysis.

Variables	Standardized ẞ	P-value
The age of gestation	-.151	.563
Maternal age	-.249	.027
Neonate birth weight	.302	.051
The width of Umbilical Cord	.210	.330

(Adjusted P=.003, R²=.146).

No competing interests reported.

Download PDF

Editorial decision: Revision requested
12 Jun, 2024
Reviews received at journal
07 Jun, 2024
Reviews received at journal
27 May, 2024
Reviewers agreed at journal
23 May, 2024
Reviewers agreed at journal
23 May, 2024
Reviewers invited by journal
23 May, 2024
Editor assigned by journal
23 May, 2024
Editor invited by journal
22 May, 2024
Submission checks completed at journal
21 May, 2024
First submitted to journal
17 May, 2024

You are reading this latest preprint version

Maternal and Neonatal Factors' Effects on Wharton's Jelly Mesenchymal Stem Cell Yield

Status:

Version 1

Abstract

Figures

Introduction

Patients and Methods

Statistical analysis:

Results

Discussion

Declarations

Competing Interests:

Author Contribution

Data Availability

References

Tables

Additional Declarations

Status:

Version 1