Wharton's Jelly Stem Cells Research Articles

The I-Guana Shot Protocol, An Intracavernosal Application of Wharton's Jelly Stem Cells for the Treatment of Erectile Dysfunction, Phase II Study

Umbilical cord has become an increasingly used source of mesenchymal stromal cells for preclinical and, more recently, clinical studies. Different clinical uses on multiple anatomical structures have demonstrated the safe use of the umbilical cord tissue and derivates with multiple potentially different options to treat difficult diseases or clinical conditions. Immunophenotypic analysis and characterization of stem cells isolated from Wharton’s jelly via various methods is currently underway in clinical research. Stem cells are the body’s basic cells from which all other cells with specialized functions are generated. Under the right conditions in the body or perinatal environment, stem cells divide to form more new cells. These new cells become stem cells (self-renewal type of cells), and can also become specialized cells (differentiated) with more specific functions, such as blood cells, brain cells, heart muscle, urinary and genital tract, bone, etc. No other cell in the body has the natural ability to generate new cell types as stem cells do. As with any emerging scientific/medical field having such incredible potential, numerous research trials and data gathering efforts are currently being conducted world-wide. New data is being reported daily that present compelling cases for therapies using certain pathologies on selected patients. Scientists have been working for decades to harness the regenerative “power” of stem cells for the possible repair and rejuvenation of certain organs to activate tissue regeneration. The ultimate objectives of all these studies are the same: deliver transformative therapies. The capacity for these cells to transform into whatever cells or tissues that the body may need to regenerate itself could, within the next 3 generations, potentially provide the means to successfully treat blindness, spinal cord injuries, diabetes, cystic fibrosis, muscular dystrophy, or even erectile dysfunction in affected patients. Such future outcomes may be possible for candidate patients who are fortunate enough to have access to those scientific and evidence- based medical protocols that use such potent biological sources. It is vital that, in the light of the dazzling potential, mesenchymal stem cells research and medical applications continue, especially for compassionate medical use (e.g. for No Option patients).

Wharton's jelly stem cells delivered via a curcumin-loaded nanofibrous wound dressings improved diabetic wound healing via upregulating VEGF and IGF genes: An in vitro and in vivo study

Diabetic wounds pose an enduring clinical hurdle, marked by delayed recovery, persistent inflammation, and an elevated susceptibility to infections. Conventional treatment approaches often fall short of delivering optimal outcomes, prompting the exploration of innovative methods to enhance the healing process. Electrospun wound dressings offer superior healing, controlled drug release, enhanced cell proliferation, biocompatibility, high surface area, and antimicrobial properties. In the current study, polycaprolactone/gelatin-based nanofibrous wound dressings were developed for the delivery of Wharton's jelly stem cells and curcumin into the diabetic wounds bed. Curcumin was loaded into the polycaprolactone/gelatin solution and electrospun to produce curcumin-loaded scaffolds. In vitro experiments including scanning electron microscopy, cell viability assay, release assay, hemocompatibility assay, cell proliferation assay, and antibacterial assay were utilized to characterize the delivery system. Then, curcumin-loaded scaffolds were seeded with 30,000 Wharton's jelly stem cells and implanted into a rat model of diabetic wounds. Study showed that the scaffolds containing both Wharton's jelly stem cells and curcumin significantly improved diabetic wound closure (86.32 3.88% at the end of 14th day), augmented collagen deposition, and improved epithelial tissue formation. Gene expression studies showed that VEGF and IGF genes were significantly upregulated by the co-delivery system. Our developed system may have augmented diabetic wound healing via upregulating pro-healing genes.

The Molecular Regulatory Mechanism in Multipotency and Differentiation of Wharton's Jelly Stem Cells.

Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) are isolated from Wharton's jelly tissue of umbilical cords. They possess the ability to differentiate into lineage cells of three germ layers. WJ-MSCs have robust proliferative ability and strong immune modulation capacity. They can be easily collected and there are no ethical problems associated with their use. Therefore, WJ-MSCs have great tissue engineering value and clinical application prospects. The identity and functions of WJ-MSCs are regulated by multiple interrelated regulatory mechanisms, including transcriptional regulation and epigenetic modifications. In this article, we summarize the latest research progress on the genetic/epigenetic regulation mechanisms and essential signaling pathways that play crucial roles in pluripotency and differentiation of WJ-MSCs.

Development of stromal differentiation patterns in heterotypical models of artificial corneas generated by tissue engineering.

Purpose: We carried out a histological characterization analysis of the stromal layer of human heterotypic cornea substitutes generated with extra-corneal cells to determine their putative usefulness in tissue engineering. Methods: Human bioartificial corneas were generated using nanostructured fibrin-agarose biomaterials with corneal stromal cells immersed within. To generate heterotypical corneas, umbilical cord Wharton's jelly stem cells (HWJSC) were cultured on the surface of the stromal substitutes to obtain an epithelial-like layer. These bioartificial corneas were compared with control native human corneas and with orthotypical corneas generated with human corneal epithelial cells on top of the stromal substitute. Both the corneal stroma and the basement membrane were analyzed using histological, histochemical and immunohistochemical methods in samples kept in culture and grafted in vivo for 12months in the rabbit cornea. Results: Our results showed that the stroma of the bioartificial corneas kept ex vivo showed very low levels of fibrillar and non-fibrillar components of the tissue extracellular matrix. However, in vivo implantation resulted in a significant increase of the contents of collagen, proteoglycans, decorin, keratocan and lumican in the corneal stroma, showing higher levels of maturation and spatial organization of these components. Heterotypical corneas grafted in vivo for 12months showed significantly higher contents of collagen fibers, proteoglycans and keratocan. When the basement membrane was analyzed, we found that all corneas grafted in vivo showed intense PAS signal and higher contents of nidogen-1, although the levels found in human native corneas was not reached, and a rudimentary basement membrane was observed using transmission electron microscopy. At the epithelial level, HWJSC used to generate an epithelial-like layer in ex vivo corneas were mostly negative for p63, whereas orthotypical corneas and heterotypical corneas grafted in vivo were positive. Conclusion: These results support the possibility of generating bioengineered artificial corneas using non-corneal HWJSC. Although heterotypical corneas were not completely biomimetic to the native human corneas, especially ex vivo, in vivo grafted corneas demonstrated to be highly biocompatible, and the animal cornea became properly differentiated at the stroma and basement membrane compartments. These findings open the door to the future clinical use of these bioartificial corneas.

Expression of Basement Membrane Molecules by Wharton Jelly Stem Cells (WJSC) in Full-Term Human Umbilical Cords, Cell Cultures and Microtissues.

Wharton's jelly stem cells (WJSC) from the human umbilical cord (UC) are one of the most promising mesenchymal stem cells (MSC) in tissue engineering (TE) and advanced therapies. The cell niche is a key element for both, MSC and fully differentiated tissues, to preserve their unique features. The basement membrane (BM) is an essential structure during embryonic development and in adult tissues. Epithelial BMs are well-known, but similar structures are present in other histological structures, such as in peripheral nerve fibers, myocytes or chondrocytes. Previous studies suggest the expression of some BM molecules within the Wharton's Jelly (WJ) of UC, but the distribution pattern and full expression profile of these molecules have not been yet elucidated. In this sense, the aim of this histological study was to evaluate the expression of main BM molecules within the WJ, cultured WJSC and during WJSC microtissue (WJSC-MT) formation process. Results confirmed the presence of a pericellular matrix composed by the main BM molecules-collagens (IV, VII), HSPG2, agrin, laminin and nidogen-around the WJSC within UC. Additionally, ex vivo studies demonstrated the synthesis of these BM molecules, except agrin, especially during WJSC-MT formation process. The WJSC capability to synthesize main BM molecules could offer new alternatives for the generation of biomimetic-engineered substitutes where these molecules are particularly needed.

The Role of Stem Cells Derived From the Mesenchyme of the Umbilical Cord in Reducing Immunosuppressive Drug Doses Used in Allogenic Transplantations.

This study evaluated the potential of Wharton's jelly mesenchymal stem cells with high tolerogenic properties in reducing immunosuppressive dosage and related adverse effects. A 4- to 6-week-old, 30-40 g weight, male inbred CD57BL/6 mice were used as skin allograft donors, whereas Balb/c mice with similar characteristics were used as recipients. Wharton's jelly stem cells were obtained from a commercial kit sourced from human umbilical cord. Skin allografts were performed from CD57Bl6 to Balb/c mice (day 0). Group 1 (control) received no treatment. Group 2 received 15 mg/kg cyclosporin A on days 0 to 30. Group 3 received 5.7 × 10 6 and 10.3 × 10 6 cell/kg Wharton's jelly stem cells on days 0 and 3, respectively. Groups 4, 5, and 6 received a combination of 15, 10, and 5 mg/kg per day cyclosporine A (days 0 to 30) with the same stem cell dose with group 3, respectively. Graft rejection was evaluated with digital photography and thermal imaging, histopathology (Banff grading, epithelialization scores, dermoepidermal dissociation), immunochemistry (Ki-67 and Bcl-2), and biochemical methods (interleukin 10, interleukin 2, interferon γ, tumor necrosis factor α) (day 10). Cumulative adverse effects of cyclosporin A occurring in the groups were revealed by histopathological evaluation of kidney and liver (a modified semiquantitative method of infiltration of inflammatory cells around the portal area and lobular region in liver; modification of the Banff rating of proximal tubules and hypertrophia of juxtaglomerular apparatus cells in kidney) (day 30). There was no rejection in groups 2, 4, and 5 until the end of study. These were statistically different versus groups 1 (day 10 ± 0.71), 3 (day 11 ± 0.82), and 6 (day 11 ± 0.58) (all P 's < 0.05). Groups 4 and 5 have exhibited statistically similar findings in histopathological (4 epithelization score: 3.7 ± 1.3; 5 epithelization score: 3.5 ± 0.5; 4 Banff grading score: 0.8 ± 0.6; 5 Banff grading score: 1.0 ± 0.5; both P 's = 1.00), immunohistochemical (4 Bcl-2 score: 3.5 ± 0.5, P = 0.618; 5 Bcl-2 score: 3.4 ± 0.5, P = 1.00; 4 Ki-67 score: 3.7 ± 0.4, P = 1.00; 5 Ki-67 score: 3.5 ± 0.5, both P 's = 1.00), and levels of cytokines (both P 's = 1.00) versus group 2. Adverse effects on kidneys and liver were lowest and statistically similar in groups 3, 5, and 6 (all P 's = 00) versus group 1. Wharton's jelly mesenchymal stem cells alter bioavailability of cyclosporine, albeit at much lower doses and with fewer systemic adverse effects.

Assessment the Effect of Human Umbilical Cord Wharton's Jelly Stem Cells on the Expression of Homing Genes: CXCR4 and VLA-4 in Cell Line of Prostate Cancer.

Background : Prostate cancer is the second most common cancer in the male that affects the health, social and economic life of person. Different compounds such as Wharton's jelly, have been used to treat prostate cancer. Wharton's jelly is a tissue rich in cells with mesenchymal morphology. Wharton's jelly compound inhibited the growth of various cancer cells, including ovarian, osteosarcoma, breast, and prostate cancers, and also reduced the expression of CXCR4 and VLA-4 genes involved in the metastasis process. Materials and Methods: To do this research, Wharton's jelly stem cells and DU145 cancer cell line were cultured. After cell culture, the effect of Wharton's jelly on this cell line was evaluated by scratching and MTT assay. The expression of CXCR4 and VLA-4 genes was also evaluated by Real-time PCR. Results: The results of MTT and Scratching tests showed that Wharton's jelly inhibited the growth of DU145 cancer cells and also decreased the expression level of CXCR4 and VLA-4 genes. Conclusion: The results of this study showed that Wharton's jelly can be considered as an effective compound for decreasing metastasis of prostate cancer.

Inhibition of Cervical Cancer Cell Migration by Human Wharton’s Jelly Stem Cells

Background: One of the most prevalent malignancies to strike women, both in Iran and globally, is cervical cancer. Metastasis, which is a significant cause of mortality, is one of the most significant pathological processes of this cancer. Therefore, preventing the migration of cancer cells may be a useful therapeutic approach. The aim of this work was to investigate the impact of conditioned media and human Wharton's jelly stem cells (hWJSCs) on the migration and growth of the cervical cancer cell line Hela as well as the in vitro mRNA expression of genes involved in metastasis. Methods: After primary culture, cellular extract and conditioned medium of hWJSCs were prepared. The viability of cervical cancer cells was investigated by MTT assay after treatment with cellular extract and conditioned medium of hWJSCs. Moreover, the anti-migratory effects of cellular extract and conditioned medium of hWJSCs on the cervical cancer cells were evaluated by wound‐healing migration assay. Finally, the mRNA expression of migration-related genes (E-cadherin and Vimentin) was detected by real-time PCR. Results: Our results indicated that the cellular extract and conditioned medium of hWJSCs (with 32% concentration) inhibited the proliferation of 100% and 20% of Hela cancer cells, respectively. In addition, the cellular extract and conditioned medium of hWJSCs significantly decreased morphological alteration and migration of the cancer cells. The cellular extract and conditioned medium of hWJSCs modified the expression of Vimentin and E-cadherin genes to inhibit the cancer cell migration (p&lt;0.05). However, the cellular extract indicated significantly profound inhibitory effects on the cervical cancer cells compared to the conditioned medium. Conclusions: Our study demonstrated that the cellular extract and conditioned medium of hWJSCs inhibit the proliferation and migration of cervical cancer cells through the modification of migration-related gene expression. However, further in vitro and in vivo studies are required for more accurate results.

Assessment the Effect of Human Umbilical Cord Wharton's Jelly Stem Cells on the Expression of Homing Genes; CXCR4 and VLA-4 in Cell Line of Breast Cancer.

Background: Breast cancer is the most common cancer in women. The prevalence of breast cancer in Western women is one in eight. Although the prevalence of breast cancer in Iran is lower than in Western countries (one in every 10-12 women), the incidence of breast cancer in it is 5-10 years earlier than in Western countries. Breast cancer is the second leading cause of cancer death among women after lung cancer. Therefore, finding new therapeutic methods could potentially help to reduce breast cancer mortality and increase the survival rate. Wharton jelly stem cells with mesenchymal morphology play an important role in inhibiting the progression of ovarian, osteosarcoma, and breast cancer by inducing apoptosis and reducing metastasis. Several environmental and genetic factors are involved in the occurrence of breast cancer. CXCR4 and VLA-4 genes are important genetic factors in breast cancer that play a role in cell survival, migration, proliferation, and metastasis of several types of cancer, especially breast cancer. Therefore, inhibition of these two genes by Wharton's Jelly Stem Cells could be a novel and effective therapeutic target in breast cancer. The aim of this study was to investigate the effect of Wharton jelly stem cells secretion on the expression of CXCR4 and VLA-4 genes in cancer cells. Materials and Methods: These cells were exposed to Wharton's Jelly Stem Cells after culturing breast cancer cells. RNA was extracted from treated cells. The expression of CXCR4 and VLA-4 genes was evaluated by real-time PCR. Results: The results of the MTT and Scratching tests showed a significant difference compared to the control group. Also, the results of Real-time PCR showed a significant decrease in the expression of CXCR4 and VLA-4 genes compared to the control group. Conclusion: The results of this study showed that different concentrations of Wharton Jelly Stem Cells reduce cancer cell growth and expression of CXCR4 and VLA-4 homing genes in MDA-MB-231 breast cancer cells. Therefore, Wharton Jelly Stem Cells can be considered as an effective treatment for breast cancer.

Intravenous Wharton's Jelly stem cell increased the number of ? cells pancreas and reduced the fasting blood glucose level in diabetes mellitus Wistar rat male (Rattus norvegicus)

Introduction: Diabetes mellitus is a metabolic disease characterized by hyperglycemic and disorders of carbohydrate, protein and fat metabolism due to abnormalities in insulin secretion, insulin sensitivity or both. Based on previous research, Wharton's Jelly's mesenchymal stem cells have anti-inflammatory, immunoregulatory properties that can improve metabolic control and have the ability to differentiate into pancreatic lineage cells that function as insulin-producing cells in vitro culture. The purpose of this study was to prove that administration of mesenchymal stem cells Wharton's Jelly can increase the number of pancreatic beta cells and reduce fasting blood glucose levels in male rats (Rattus norvegicus) Wistar strain of diabetes mellitus.Methods: The research design used was pure experimental with post-test only control group design using 36 white male rats. All samples were induced with Streptozotocin and Nicotinamide. The selected samples were divided into two groups: the control group given glibenclamide + 0.9% NaCl, and the treatment group was given glibenclamide + mesenchymal stem cells from Wharton's Jelly. Calculation of the number of pancreatic beta cells and measurement of blood glucose levels were carried out after 14 days of treatment.Result: The results showed that the treatment group had a higher number of pancreatic beta cells than the control group (105.17 ± 16.379 cells/field of view vs. 54.00 ± 11.366 cells/field of view) (p &lt;0.001). In addition, the treatment group had lower fasting blood glucose levels than the control group (109.06 ± 16.71 mg/dl vs 122.78 ± 10.14 mg / dl) (p &lt;0.05).Conclusion: It was concluded that intravenous administration of Wharton's Jelly mesenchymal stem cells increased the number of pancreatic beta cells and decreased fasting blood glucose levels in male rats (Rattus norvegicus) Wistar strain of diabetes mellitus.

Wharton jelly stem cells inhibits AGS gastric cancer cells through induction of apoptosis and modification of MAPK and NF-κB signaling pathways

BackgroundGastric cancer) GC) is one of the most common cancer with high mortality worldwide. The human Wharton's jelly stem cells (hWJSCs) can inhibit several cancer cells through several molecular pathways. Therefore, the present study aimed to investigate anticancer effects of hWJSCs conditioned medium (hWJSC-CM) and cell-free lysate (hWJSC-CL) against of GC cell line AGS and underlying signaling pathways. MethodsIn this study, we evaluated the effects of hWJSC-CM and hWJSC-CL on viability, proliferation, migration, invasion, apoptosis, and MAPK and NF-κB signaling pathways in AGS cells. Moreover, mRNA expression of genes involved in apoptosis (BAX, BCL2, SMAC, and SURVIVIN), as well as expression of proteins involved in NF-κB and MAPK signaling pathways were evaluated. ResultsThe obtained results showed that the hWJSC-CM and hWJSC-CL decreased viability, migration, and invasion of GC cell line AGS in a concentration and time dependent manner. We observed that the hWJSC-CM and hWJSC-CL induced apoptosis pathway through regulation of apoptosis involved genes mRNA expression. In addition, the hWJSC-CM and hWJSC-CL suppressed NF-κB signaling pathways as well as promoted MAPK signaling pathways. ConclusionIn general, our study suggested that the hWJSC-CM and hWJSC-CL inhibits proliferation and viability of GC cell line AGS through induction of apoptosis, as well as modification of NF-κB and MAPK signaling pathways.

The Radio-Sensitivity Effects of Human Umbilical Cord Wharton’s Jelly Mesenchymal Stem Cell’s Conditional Medium on MDA-MB-231 Cells

Background: While radiotherapy is the important modality in the treatment of breast cancer cells, radioresistance of some tumor cell lines such as MDA-MB-231 is still a limitation that must be considered. Objectives: The present study was done to examine the effect of the conditioned medium of the human umbilical cord Wharton's jelly stem cells (hWJSCs + CM) on the radiosensitivity of MDA-MB-231 cells in combination with megavoltage-radiations. Methods: Groups are Control, CM, GY, and GY + CM. In irradiation groups, breast cancer cells were exposured with 4, 6, and 8 Gy radiation. Each group includes different doses of the conditioned medium of hWJSCs (25%, 50%, and 75%). Results: The MTT assay showed that the proliferative activity of Gy + CM groups at all doses of condition medium decreased significantly compared with the control, rather than Gy groups. Trypan blue viability test showed that the survival rate of MDA-MB-231 cells significantly reduced in the CM and 8 Gy + CM groups compared with the control group, rather than Gy groups. MDA-MB-231 cells lost their normal spindle shape and became thinner and longer after 48h of treatment and the number of cells sharply reduced in Gy + CM groups compared with the control group, rather than Gy groups. These changes were accompanied by inducing significant up-regulation of Interleukin-6 (IL-6) in the 4 Gy + CM and 8 Gy + CM groups compared with the control group, rather than Gy groups and as a consequence, a decrease in the amount of transmembrane tumor necrosis factor-α (tmTNF-α) as a pro-inflammatory cytokine in the Gy + CM groups compared with the control group, rather than Gy groups. Also, we indicated that the radiosensitivity of breast cancer cells was probably enhanced by an increase in different doses of the conditioned medium of stem cells. Conclusions: Treatment of the MDA-MB-231 cells with hWJSCs + CM plus radiotherapy inhibited the growth and proliferation of cancer cells and this method is a novel strategy for breast cancer therapy by overcoming radioresistance.

Tissues derived from reprogrammed Wharton's jelly stem cells of the umbilical cord as a platform to study gestational diabetes mellitus.

Gestational diabetes mellitus (GDM) has been strongly associated with an increased risk of type 2 diabetes mellitus (T2DM) in later child and adulthood. The human umbilical cord and its contents are of fetal origin and represent the fetus genetically and physiologically. Since it is not possible to obtain tissues from the fetus and newborn to investigate the association between GDM and later T2DM, we reprogrammed the stem cells from the Wharton's jelly of umbilical cords (hWJSCs) of GDM and non-GDM mothers into induced pluripotent stem cells (iPSCs) and then differentiated the iPSCs into insulin-producing cells (IPCs) to provide pancreatic tissues that represent the fetus of GDM and normal mothers. These tissues are an attractive model to study the effects of glucose on the fetus. Interestingly, GDM-iPSCs had a decreased potential towards differentiation into IPCs. IPCs differentiated from GDM-iPSCs also had lower total insulin content and a lower capacity for insulin secretion to glucose stimulation compared to their normal-iPSC counterparts. This abnormal pathogenesis in GDM-iPSCs pancreatic differentiation recapitulates the pathology that may be observed in the infants of the diabetic mother (IDM) and while indicating adaptive mechanisms for fetal survival, may lead to the development of T2DM later in life. (199 words).

Biosynthesis, characterization and evaluation of the supportive properties and biocompatibility of DBM nanoparticles on a tissue-engineered nerve conduit from decellularized sciatic nerve.

In this study, we examined the supporting effects of nano-demineralized bone matrix on the cultivation of Wharton's jelly stem cells on acellularized nerve scaffold. Demineralized bone matrix nanoparticles were prepared and characterized by several experiments. Decellularized sciatic nerve scaffolds were prepared and their efficiency was evaluated using histological stainings and biomechanical testing. Results of histological staining indicated that the integrity of the extra cellular matrix components was preserved. Also, the growth and viability of WJSCs on the scaffolds were significantly higher in DBM nanoparticle groups. We conclude that supportive properties of nano-DBM groups showed better cell viability and a suitable microenvironment for proliferation, retention, and adhesion of cells compared with other groups.

Human Wharton's jelly stem cells inhibit endometriosis through apoptosis induction.

Endometriosis is a relatively benign disease characterized by endometrial tumors and uterus stroma. Apoptosis suppression is one of the most important pathological processes of endometriosis. Recently, several studies reported that human Wharton's jelly stem cells (hWJSCs) can inhibit growth and proliferation of various cancer cells through induction of apoptosis. Therefore, the aim of the present study was to investigate the effects of hWJSCs conditioned medium (hWJSC-CM) and cell-free lysate (hWJSC-CL) on endometriosis cells in vitro. In the present study, effects of different concentrations of hWJSC-CM and hWJSC-CL on viability and proliferation, morphological alterations, colony formation, migration, invasion, and apoptosis of endometriosis cells were evaluated. Our results showed that hWJSC-CM and hWJSC-CL decrease viability and proliferation, colony formation, migration, and invasion, as well as increase morphological alterations and apoptosis of endometriosis cells, in a concentration- and time-dependent manner. Decreased migration and invasion of treated endometriosis cells with hWJSC-CM and hWJSC-CL may be due to decrease of MMP-2 and MMP-9 gene expression. Moreover, induction of apoptosis in treated endometriosis cells can be due to regulation of apoptosis-related genes expression, including BAX, BCL-2, SMAC, and SURVIVIN. The results of the present study suggest that hWJSC-CM and hWJSC-CL can inhibit endometriosis cells at a mild-to-moderate level through various physiological mechanisms. However, further studies on animal models are necessary to achieve more accurate results.

Hypoxic Wharton's Jelly Stem Cell Conditioned Medium Induces Immunogenic Cell Death in Lymphoma Cells.

Mesenchymal stem cells from Wharton's jelly of the human umbilical cord (hWJSCs), and the conditioned medium (hWJSC-CM) prepared from them, were shown to be tumoricidal on many cancers. However, these tumoricidal effects were observed in hWJSCs grown under normoxic conditions of 21% oxygen in the laboratory. Since oxygen concentrations in the stem cell niche or physiological microenvironment are hypoxic and help to maintain stemness properties, the objective of this work was to evaluate whether there were differences in the tumoricidal properties of hWJSC-CM grown in 21% O2 (normoxic) or 5% O2 (hypoxic) environments. The results showed that hWJSCs grown under normoxic or hypoxic conditions showed no distinct morphological differences in culture and remained positive in trilineage differentiation into adipocytes, osteocytes, and chondrocytes. Hypoxic hWJSCs expressed the mesenchymal stem cell surface markers CD105, CD90, CD73, CD146, and CD108 similar to normoxic hWJSCs but were negative for the hematopoietic markers CD14, CD19, CD34, CD45, CD117, and HLA-DR. Hypoxic hWJSC-CM produced a significantly greater reduction in cell viability and a significantly greater increase in apoptosis, oxidative stress, and lipid peroxidation in human lymphoma cells compared to normoxic hWJSC-CM. Hypoxic hWJSC-CM also produced significantly greater expression of immunogenic cell death (ICD) hallmarks such as surface-bound calreticulin, HSP70, HSP90, and high mobility group binding 1 proteins and significantly decreased expression of the defense molecules CD47 and PD-L1. This study showed that the tumoricidal effect of hypoxic hWJSC-CM was superior to normoxic hWJSC-CM and should be the preferred choice of preparing hWJSC-CM for the induction of ICD on lymphoma cells.

Induction of HLA-B27–Associated Reactive Arthritis After a Wharton’s Jelly “Stem Cell” Injection

Orthobiologic injections are an emerging treatment option for musculoskeletal conditions. Allogenic gestational tissue-derived products, including umbilical cord-derived Wharton's jelly, are among the available options. These tissues are currently being harvested and sold by companies as an off-the-shelf "stem cell" injection for conditions such as back pain and osteoarthritis. Despite documented infections from similar umbilical cord derivatives, "stem cell clinics" continue to use them. Here, we report the first documented case of HLA-B27-associated reactive arthritis induced by a Wharton's jelly stem cell injection. A 36-yr-old man was injected with Wharton's jelly for low back pain and within 24 hours developed fevers, chills, polyarthritis, and enthesitis. Infectious disease work-up was negative. Inflammatory markers were elevated and his HLA-B27 antigen was positive. Initial treatment included methylprednisolone and sulfasalazine. This case highlights the unknown dangers of these allogenic injections and physicians should remain cautious about their use until further study and regulation can ensure patient safety.

Expanded Differentiation Capability of Human Wharton's Jelly Stem Cells Toward Pluripotency: A Systematic Review.

Human Wharton's jelly stem cells (HWJSC) can be efficiently isolated from the umbilical cord, and numerous reports have demonstrated that these cells can differentiate into several cell lineages. This fact, coupled with the high proliferation potential of HWJSC, makes them a promising source of stem cells for use in tissue engineering and regenerative medicine. However, their real potentiality has not been established to date. In the present study, we carried out a systematic review to determine the multilineage differentiation potential of HWJSC. After a systematic literature search, we selected 32 publications focused on the differentiation potential of these cells. Analysis of these studies showed that HWJSC display expanded differentiation potential toward some cell types corresponding to all three embryonic cell layers (ectodermal, mesodermal, and endodermal), which is consistent with their constitutive expression of key pluripotency markers such as OCT4, SOX2, and NANOG, and the embryonic marker SSEA4. We conclude that HWJSC can be considered cells in an intermediate state between multipotentiality and pluripotentiality, since their proliferation capability is not unlimited and differentiation to all cell types has not been demonstrated thus far. These findings support the clinical use of HWJSC for the treatment of diseases affecting not only mesoderm-type tissues but also other cell lineages. Impact statement Human Wharton's jelly stem cells (HWJSC) are mesenchymal stem cells that are easy to isolate and handle, and that readily proliferate. Their wide range of differentiation capabilities supports the view that these cells can be considered pluripotent. Accordingly, HWJSC are one of the most promising cell sources for clinical applications in advanced therapies.

Inhibition of Cervical Cancer Cell Line Hela by Human Wharton’s Jelly Stem Cells Through Induction of Apoptosis

Background: Cervical cancer is one of the most common cancers of women in the world, which causes high mortality. The human umbilical cord Wharton's jelly stem cells (hWJSCs) can inhibit various cancer cells. Objectives: This study aimed to investigate the effects of conditioned medium and cellular extract of human umbilical cord hWJSCs on cervical cancer cell line, Hela. Methods: After isolation and primary culture of hWJSCs, conditioned medium and cellular extracts of hWJSCs were prepared, and its anti-proliferative effects were evaluated on cervical cancer cells, Hela using micro-culture tetrazolium (MTT) assay. After total RNA extraction and cDNA synthesis, expression of apoptosis-related BCL-2 and BAX genes were evaluated using real-time PCR. Results: The results showed that conditioned medium (55% concentration in 72 hours) and cellular extraction (10% concentration in 24 hours) caused death of 50% cancer cells (IC50). The anti-cancer effects of conditioned medium and cellular extraction were concentration- and time-dependent. The conditioned medium and cellular extract of hWJSCs significantly down-regulated and up-regulated mRNA expression of apoptosis-related BCL-2 and BAX genes, respectively. Conclusions: Our study showed that conditioned medium and cellular extract of human umbilical cord hWJSCs inhibit viability and proliferation of cervical cancer cells. However, further studies on animal models are necessary for more accurate results.

The effect of allogenic human Wharton's jelly stem cells seeded onto acellular dermal matrix in healing of rat burn wounds.

Various methods were introduced to overcome the autograft shortage in burn wound care, including cell transplantation and tissue engineering. To evaluate the healing effect of allogenic human Wharton's jelly stem cells (hWJSCs) seeded onto acellular dermal matrix (ADM) in rat burn injuries. Human Wharton's jelly stem cells provided from umbilical cord tissue were characterized before transplantation, and the growth kinetic was determined. Skin samples from cosmetic surgeries were used for preparation of ADM. Forty male Sprague Dawley rats were randomly divided into 4 equal groups. Third-degree burn was induced for all animals by exposing to hot water using a 2cm ring for 10seconds. Group 1 was burned rats that did not receive any treatment. After burn injury, the second group received silver sulfadiazine (SSD), the third group was treated just by using ADM, and the fourth group received 2×106 hWJSCs seeded onto ADM. The animals were euthanized for histologic evaluation after 7, 14, and 21days. Human Wharton's jelly stem cells were characterized to be spindle shape and positive for osteogenic and adipogenic induction and for mesenchymal markers but lacked hematopoietic markers. Population doubling time (PDT) was 40.1hours with an increasing growth trend until day 6th. Macro- and microscopically, the healing was mild in ADM group and moderate in ADM+hWJSCs group after 21days. Allogenic hWJSCs seeded onto ADM improved the healing process in burn wounds denoting to their therapeutic and anti-inflammatory effects in burn wounds that can be added to the literature.