Breast Milk Stem Cells Research Articles

Breast Milk Stem Cells in the Treatment of Neonatal Diseases

Abstract:: Breast milk plays an important role in preventing and remission various acute and chronic neonatal diseases. However, the mechanisms through which breast milk performs these effects are still unclear. Recently, research has shown that breast milk contains previously undiscovered cells, called breast milk stem cells (BMSCs), which have the potential for multiple differentiation. These cells have been shown to be expressed in multiple organs and systems of infants after ingestion. This study found that newborns who received BMSCs required a shorter treatment duration and had a better prognosis than those who did not receive BMSCs. This suggests that BMSCs may be able to treat a variety of neonatal diseases, and breast milk may play a role in preventing and mitigating neonatal diseases through BMSCs. In this paper, we summarized the research progress on BMSCs and their therapeutic effects on organs, the possible ways in which BMSCs enter different organs and tissues of the human body, and detailed the types of diseases, modes of action, and mechanisms that BMSCs may play a role in. We aim to pave the way for further clinical and preclinical research on BMSCs, as well as the treatment of neonatal diseases with BMSCs, to provide possible new therapeutic means for treating neonatal diseases and a variety of other diseases.

Evaluation puramatrix as a 3D microenvironment for neural differentiation of human breastmilk stem cells

The extracellular matrix is recognized as an efficient and determining component in the growth, proliferation, and differentiation of cells due to its ability to perceive and respond to environmental signals. Applying three-dimensional scaffolds can create conditions similar to the extracellular matrix and provide an opportunity to investigate cell fate. In this study, we employed the PuraMatrix hydrogel scaffold as an advanced cell culture platform for the neural differentiation of stem cells derived from human breastmilk to design an opportune model for tissue engineering.Isolated stem cells from breastmilk were cultured and differentiated into neural-like cells on PuraMatrix peptide hydrogel and in the two-dimensional system. The compatibility of breastmilk-derived stem cells with PuraMatrix and cell viability was evaluated by scanning electron microscopy and MTT assay, respectively. Induction of differentiation was achieved by exposing cells to the neurogenic medium. After 21 days of the initial differentiation process, the expression levels of glial fibrillary acidic protein (GFAP), microtubule-associated protein (MAP2), β-tubulin III, and neuronal nuclear antigen (NeuN) were analyzed using the immunostaining technique.The results illustrated a notable expression of MAP2, β-tubulin-III, and NeuN in the three-dimensional cell culture in comparison to the two-dimensional system, indicating the beneficial effect of PuraMatrix scaffolds in the process of differentiating breastmilk-derived stem cells into neural-like cells.In view of the obtained results, the combination of breastmilk-derived stem cells and PuraMatrix hydrogel scaffold could be an advisable preference for neural tissue regeneration and cell therapy.

Human breast milk cells are positive for the pioneer transcription factor ISL1.

ISL1 is a pioneer transcription factor that plays important roles in cell lineage specification and differentiation, by programming the epigenome and recruiting additional regulatory factors. The aim of this study is to determine whether the human breastmilk contains ISL1-positive stem cells, and, if so, to describe the subcellular localization of ISL1. Breast milk was obtained from fourteen healthy females during the first 2-6 months of lactation. Cell morphology was examined in the breast milk with the automatic ThinPrep® processor (Hologic® Inc.) in commercial Cytological ThinPrep® solution (Hologic® Inc.), followed by standard immunohistochemical staining of ISL1. ISL1 had a granular diffuse cytoplasmic localization, with varying intensity of staining in both single and grouped cells. Nuclear staining was also present, as was staining of intracellular and extracellular vesicles with ISL1 antibody. These preliminary results suggest that ISL1 could distinguish a readily available source of putative stem cells in human breast milk. These stem cells may complete the network created between the mother and the newborn during gestation, thereby improving the efficiency of programming and reprogramming postnatal events.

Potential Epigenetic Effects of Human Milk on Infants' Neurodevelopment.

The advantages of human milk feeding, especially in preterm babies, are well recognized. Infants' feeding with breast milk lowers the likelihood of developing a diverse range of non-communicable diseases later in life and it is also associated with improved neurodevelopmental outcomes. Although the precise mechanisms through which human milk feeding is linked with infants' neurodevelopment are still unknown, potential epigenetic effects of breast milk through its bioactive components, including non-coding RNAs, stem cells and microbiome, could at least partly explain this association. Micro- and long-non-coding RNAs, enclosed in milk exosomes, as well as breast milk stem cells, survive digestion, reach the circulation and can cross the blood-brain barrier. Certain non-coding RNAs potentially regulate genes implicated in brain development and function, whereas nestin-positive stem cells can possibly differentiate into neural cells or/and act as epigenetic regulators in the brain. Furthermore, breast milk microbiota contributes to the establishment of infant's gut microbiome, which is implicated in brain development via epigenetic modifications and key molecules' regulation. This narrative review provides an updated analysis of the relationship between breast milk feeding and infants' neurodevelopment via epigenetics, pointing out how breast milk's bioactive components could have an impact on the neurodevelopment of both full-term and preterm babies.

Antiviral properties of breast milk: a new area of research

Human milk uniquely combines nutritional, immunological, enzymatic and hormonal properties. The properties of secretory IgA (sIgA) have been studied for more than half a century, the role of oligosaccharides has been studied for more than 30 years, and the importance of breast milk stem cells has been discussed for more than 10 years. Wide interest, including in connection with SARS-CoV-2-, HIV-, rota-, norо-, herpes- and other viral infections, was caused by various (more than 10 types) protective factors with antiviral activity present in human milk. This review discusses the properties and significance of immunoglobulins, lactoferrin, oligosaccharides, lysozyme, cytokines as antiviral factors of both scientific and practical importance.

Exploring cell surface markers and cell-cell interactions of human breast milk stem cells.

Breakthrough studies have shown that pluripotent stem cells are present in human breast milk. The expression of pluripotency markers by breast milk cells is heterogeneous, relating to cellular hierarchy, from early-stage multi-lineage stem cells to fully differentiated mammary epithelial cells, as well as weeks of gestation and days of lactation. Here, we qualitatively analyze cell marker expression in freshly isolated human breast milk cells, without any manipulation that could influence protein expression. Moreover, we use electron microscopy to investigate cell-cell networks in breast milk for the first time, providing evidence of active intercellular communication between cells expressing different cellular markers. The immunocytochemistry results of human breast milk cells showed positive staining in all samples for CD44, CD45, CD133, and Ki67 markers. Variable positivity was present with P63, Tβ4 and CK14 markers. No immunostaining was detected for Wt1, nestin, Nanog, OCT4, SOX2, CK5, and CD34 markers. Cells isolated from human breast milk form intercellular connections, which together create a cell-to-cell communication network. Cells freshly isolated form human breast milk, without particular manipulations, show heterogeneous expression of stemness markers. The studied milk staminal cells show "pluripotency" at different stages of differentiation, and are present as single cells or grouped cells. The adjacent cell interactions are evidenced by electron microscopy, which showed the formation of intercellular connections, numerous contact regions, and thin pseudopods.

Isolation and culture methods of human breastmilk stem cells: A systematic review

With the rapid development of cell-based therapy, the source of stem cells derived from non-invasive techniques have been intensively studied for their potential to replace the invasive source of stem cells. One of the alternative sources of stem cells is the cells taken from human breastmilk. Human breastmilk is always readily available, easy to access and can last for an extended period. Unfortunately, the standard protocol to isolate the cells from breastmilk is still not well explained. Our study aimed to investigate three main questions in human breastmilk stem cells: the optimal lactation stage, isolation and culture method, and the characterization of human breastmilk stem cells. We searched in vitro studies that used human breastmilk as a cell source of stem cells and clearly explained the step-by-step procedure. From the initial 457 studies, 15 articles fulfilled the criteria and were further analyzed. The results showed that the stem cells could be taken from breastmilk in any lactation stage and isolated with various techniques to obtain both pluri- and multipotent stem cells. Although breastmilk can be used as an alternative for noninvasive source of stem cell, the results of few study groups were found sometimes contradicting another group, resulting in confusing theory. Therefore, the standard protocol is still needed to tackle the confounding results.

Breast milk stem cells: Are they magic bullets in neonatology?

Each mammal produces milk specific to its newborn that meets all nutritional needs. Breast milk is not only a secretory product but is also a complex liquid containing several components that provide enteral nutrition. The stage of lactation, the fullness of the breast, the feeding of the baby, and the health of the mother during the breastfeeding period cause differences in the composition of breast milk. Although the positive effects of breast milk on the physical and intellectual development of a child in the short and long term have been known for centuries, its mechanism has not been elucidated. Stem cells are defined as the cells that possess specific markers and have not undergone differentiation. Under suitable conditions and stimuli, they can differentiate into desired cells. The detection of stem cells, whose exact origin is not known, in breast milk and their demonstration in the baby's body have prompted the necessity of exploring the possible role of stem cells in the treatment of diseases. In this review, breast milk-derived stem cells and their possible role in neonatology are discussed.

Proteomic analysis of hypoxia and non-hypoxia secretome mesenchymal stem-like cells from human breastmilk.

IntroductionBreastmilk contains proteins and cells which have stem cell properties. The human breastmilk stem cell mimick mesenchymal stem cells and expresses pluripotency genes. The protein level of breastmilk is high in colostrum and gradually subsides in the first year of lactation. The mesenchymal stem cells from breastmilk can be an alternative source of stem cells that can potentially affect cardiovascular therapy. This study aimed to identify the proteomic analysis of secretome mesenchymal stem-like cells under hypoxia compared to non-hypoxia from human breastmilk stem cells.Material and methodsThe human breastmilk was collected from six healthy breastfeeding women and transported to the laboratory under aseptic conditions. The breastmilk cells were isolated then cultured. After 72 h, the human breastmilk stem cells reached confluence then cleaned up and isolated in serum-free media (spheroid) to allow serial passaging every 48 h. The acquisition stem cell was made with flow cytometry. The cells were divided into hBSC secretomes under hypoxia (A) and non-hypoxia (B) and analyzed for LC-MS to identify the peptide structure.ResultsThe human breastmilk cells contained several mesenchymal stem-like cells in density 2.4 × 106 cell/mL for hypoxia and 2 × 106 cell/mL for non-hypoxia conditions. The human breastmilk stem cell surface markers derived from the third cell passage process were 93.77% for CD44, 98.69% for CD73, 88.45% for CD90, and 96.30% for CD105. The protein level of secretome mesenchymal stem -like cells under hypoxia was measured at 5.56 μg/mL and 4.28 μg/mL for non-hypoxia. The liquid chromatography-mass spectrometry analysis identified 130 and 59 peptides from hypoxia and non-hypoxia of the human breastmilk stem cell secretome sequentially. Some important proteomics structures were found in the hypoxic human breastmilk stem cell secretome, such as transforming growth factor-β, VE-cadherin, and caspase.ConclusionThe human breastmilk cells contain mesenchymal stem-like cells and a high concentration of CD44, CD73, CD90, and CD105 as surface markers at third passage culture. The hypoxic hBSC secretome produces a higher protein level compare to non-hypoxia. The transforming growth factor -β was found in the hypoxic hBSC secretome as a modulator of VEGF-mediated angiogenesis.

How Knowledge about Stem Cells Influences Attitudes towards Breastfeeding: Case Study of Polish Women.

Maternal breastfeeding is associated with multiple health benefits, both for the child and the mother. Since breastfeeding rates are declining, finding new, future-oriented strategies to strengthen and support mothers’ positive attitudes towards breastfeeding need to be developed. In this paper, we describe how information about the presence of stem cells in breast milk can influence the willingness to breastfeed in the population of Polish pregnant women. A cross-sectional study involving a group of 150 pregnant women was conducted to assess the correlation between their knowledge about stem cells and their attitude towards breastfeeding. Among the respondents, only 6% claimed that they did not know anything about stem cells, but general knowledge about stem cells in the research group was poor. The survey results indicated that city residence, university degree, maternal experience and advanced pregnancy correlated with higher general knowledge regarding stem cells. Most respondents (77.3%) had no knowledge regarding the presence of stem cells in breast milk. Approximately two-thirds of mothers with earlier negative breastfeeding experience declared that information about the presence of stem cells in breast milk could have influenced the decision to continue and extend the time of breastfeeding. Hence highlighting the presence of stem cells in breast milk can be used to encourage breastfeeding as a unique activity.

Amniotic fluid and breast milk: a rationale for breast milk stem cell therapy in neonatal diseases.

Amniotic fluid and breast milk play important roles in structural development throughout fetal growth and infancy. Given their significance in physical maturation, many studies have investigated the therapeutic and protective roles of amniotic fluid and breast milk in neonatal diseases. Of particular interest to researchers are stem cells found in the two fluids. These stem cells have been investigated due to their ability to self-replicate, differentiate, reduce tissue damage, and their expression of pluripotent markers. While amniotic fluid stem cells have received some attention regarding their ability to treat neonatal diseases, breast milk stem cells have not been investigated to the same extent given the recency of their discovery. The purpose of this review is to compare the functions of amniotic fluid, breast milk, and their stem cells to provide a rationale for the use of breast milk stem cells as a therapy for neonatal diseases. Breast milk stem cells present as an important tool for treating neonatal diseases given their ability to reduce inflammation and tissue damage, as well as their multilineage differentiation potential, easy accessibility, and ability to be used in disease modelling.

Breast Milk Stem Cell Survival in Neonate's Gut, Entery into Neonate Circulation and Adaption by the Body.

The stem cell exchange during pregnancy is thought to remain chimeras for life. Few studies recently revealed that maternal transfer of viable stem cells to the offspring continues even after birth during breastfeeding. Some of these stem cells are likely to be integrated into different organs (brain, blood, kidneys, and pancreas) including neurons and insulin-producing cells in the pancreas to become functional cells. This finding opens a new avenue for research on therapeutic uses of breast milk- derived stem cells. Recently Dr. Foteini Hassiotou used glowing mice, which were genetically modified to express a gene called tdTomato that causes cells to appear red under fluorescence light. These mice were mated, and their babies were swapped with the pups of another, unmodified mother mouse. The new pups suckled the modified mouse and, as a result, obtained glowing red stem cells from breast milk. The study has never been replicated in humans, so it is not clear yet if the findings apply to humans as in the case of mice. However, the results of the study are the jumping-off points for future research on human breast milk stem cells and their possible application in stem cell therapies. Additional studies are necessary to understand the passage of human breast milk stem cells through the neonate's GI tract, and passage to the systemic circulation.

In vitro differentiation capacity of human breastmilk stem cells: A systematic review.

BACKGROUNDMesenchymal stem cells are pluripotent cells that have the ability to generate cells from a cell line or in other cell types from different tissues but from the same origin. Although those cells have more limited differentiation capacity than embryonic stem cells, they are easily obtained from somatic tissue and can be grown in large quantities. This characteristic of undifferentiated stem cells differentiating into different cell lines arouses strategies in regenerative medicine for the treatment of different diseases such as neurodegenerative diseases.AIMTo evaluate the cell differentiation capacity of human breastmilk stem cells for the three germ layers by a systematic review.METHODSThe searched databases were PubMed, EMBASE, OVID, and COCHRANE LIBRARY, published between 2007 and 2018 in the English language. All were in vitro studies for analysis of the "cell differentiation potential" in the literature using the keywords “human breastmilk,” “stem cells,” and keywords combined with the Boolean operator “NOT” were used to exclude those articles that had the word “CANCER” and their respective synonyms, which were previously consulted according to medical subject heading terms. PRISMA 2009 guidelines were followed in this study.RESULTSA total of 315 titles and abstracts of articles were examined. From these, 21 were in common with more than one database, leaving 294 articles for analysis. Of that total, five publications met the inclusion criteria. When analyzing the publications, it was demonstrated that human breastmilk stem cells have a high cellular plasticity, exhibiting the ability to generate cells of all three germ layers, endoderm, mesoderm, and ectoderm, demonstrating their stemness. Those cells expressed the genes, TRA-1-60/81, octamer-binding transcription factor 4, and NANOG, of which NANOG, a critical regulator for self-renewal and maintenance, was the most highly expressed. Those cells have the ability to differentiate in vitro into adipocytes, chondrocytes, osteocytes, oligodendrocytes, astrocytes, and neurons as well hepatocytes, β-pancreatic cells, and cardiomyocytes.CONCLUSIONAlthough the literature has been scarce, the pluripotentiality of these cells represents great potential for tissue engineering and cellular therapy. Further studies for safe clinical translation are needed.

The Therapeutic Potential of Conditioned Medium from Human Breast Milk Stem Cells in Treating Spinal Cord Injury.

Study DesignExperimental animal study.PurposeThis study investigated the therapeutic effects of human breast milk stem cell (BMSC)-conditioned medium (BMSC-CM) in a model of spinal cord injury (SCI) in male Sprague-Dawley rats.Overview of LiteratureSCI is one of the leading causes of disability in addition to sensory and motor impairment. So far, there have been no successful treatments for SCI. Given the positive outcomes associated with using stem cells and their derivatives as a treatment for various diseases, there is a growing interest in using them as an SCI treatment. Recent research has demonstrated that CM from stem cells has therapeutic advantages.MethodsHuman BMSCs were isolated and characterized, and CM was subsequently collected. Animals received an intrathecal administration of BMSC-CM after SCI. The activity of caspase-3 was measured to assess apoptosis, and levels of tumor necrosis factor-α and interleukin-1β were measured to assess inflammation. Also, sensory and locomotor performances were assessed after SCI and BMSC-CM administration.ResultsAdministration of CM from BMSC reduced apoptosis and inflammation at the site of injury in a rat model of SCI (p<0.05). Motor, sensory, locomotor, and sensorimotor performances were significantly improved in rats that received BMSC-CM after SCI.ConclusionsIntrathecal administration of BMSC-CM improved recovery in a rat model of SCI.

Comparison and culturing different types of cells from fresh breast milk with different culture medium

Background: To investigate the properties of breast milk cells at different stages and culture different types of cells from fresh breast milk. Methods: Cell concentration and viability were analyzed after being isolated from fresh breast milk. Cell properties were tested by flow cytometry with different cell surface markers. Different types of breast milk cells were cultured with different medium. After being purified, the cells were identified by flow cytometry and immunofluorescence. Results: The concentration of breast milk cells decreased with lactation time. The viability of breast milk cells decreased gradually in both colostrum and transitional milk (C and T milk) group and mature milk group, and there was no significant difference in two groups over time. The expression levels of different cell surface markers were higher in C and T milk than mature milk. Using three different culturing media, we could get different types of cells including immune cells, mammary epithelial cells (MECs), mesenchymal stem cells (MSCs) and breast milk stem cells (BSCs) with different efficiency between C and T milk and mature milk. Conclusions: C and T milk contains more cells than mature milk, especially immunocytes. It is feasible to culture MECs with F medium, MSCs with M medium or F medium, and BSCs with CM medium.

Novel DRAQ5™/SYTOX® Blue Based Flow Cytometric Strategy to Identify and Characterize Stem Cells in Human Breast Milk.

Human breast milk could be an important stem cell source for the development of newborn and preterm infants, but quantitative data on the stem cell content in breast milk at various gestational stages are needed to determine the clinical value of breast milk as a source of stem cells. Breast milk also contains milk fat globules, lipid droplets of different sizes, debris and dead cells and these components hamper flow cytometry analysis of human breast milk samples. Here, we originally used standard protocols for flow cytometry to characterize cell populations in human breast milk but failed to discriminate between cells and noncellular components. We then applied a centrifugation protocol to separate cream and skim milk from the cell-containing pellet and used a novel staining protocol with DRAQ5™ and SYTOX® blue dye as well as antibodies to characterize cells within the pellet fraction. Flow cytometry analysis identified viable DRAQ5™+ /SYTOX® Blue- cells and determined the content of CD11b+ monocytes and TRA-1-81+ putative stem cells in human breast milk samples. Hence, we developed a novel and reliable flow cytometry based-approach to quantify subpopulation of cells in human breast milk with a high content of milk fat globules, lipid droplets, and particles. This approach will improve the identification and quantification of breast milk cells and allow standardizing the flow cytometry-based evaluation of the stem cell content. © 2018 International Clinical Cytometry Society.

Transfer and Integration of Breast Milk Stem Cells to the Brain of Suckling Pups

Beside its unique nutritional content breast milk also contains live cells from the mother. Fate of these cells in the offspring has not been adequately described. In this study, we aimed to detect and identify maternal cells in the suckling’s blood and the brain. Green fluorescent protein expressing transgenic female mice (GFP+) were used as foster mothers to breastfeed wildtype newborn pups. One week and two months after the birth, blood samples and brains of the sucklings were analyzed to detect presence of GFP+ cells by fluorescence activated cell sorting, polymerase chain reaction and immunohistochemistry on the brain sections and optically cleared brains. The tests confirmed that maternal cells were detectable in the blood and the brain of the pups and that they differentiated into both neuronal and glial cell types in the brain. This phenomenon represents breastfeeding – induced microchimerism in the brain with functional implications remain to be understood.

Stem cells in breast milk

Stem cells in breast milk

Progress of breast milk stem cells

Stem are original undifferentiated biological that have multi-directional differentiation potential and self-reproduction ability.In recent years, the research on composition of milk suggests that a new type of exist in milk.They are able to differentiate into specialized and reproduce themselves.So, they are called breast milk stem cells . Through feeding, these stem can enter a baby′s body.They not only facilitate development of the baby′s nervous system and other tissues and organs, but also provide regenerative medicine with a new treatment approach by virtue of its multi-directional differentiation potential. Key words: Breast milk; Stem cell; Regenerative medicine

Differentiation Potential of Breast Milk-Derived Mesenchymal Stem Cells into Hepatocyte-Like Cells.

Human breast milk stem cells (hBSCs) contain a population of cells with the ability to differentiate into various cell lineages for cell therapy applications. The current study examined the differentiation potential of hBSCs into hepatocytes-like cells. The cells were isolated from the breast milk and were treated with hepatogenic medium containing hepatocyte growth factor, insulin-like growth factor and dexamethasone for 7days subsequently; Oncostatin M was added to the culture media. RT-PCR and immunocytochemistry were performed to detect the hepatogenic markers. The glycogen storage and the ability of the cells to absorb and release indocynanin green were also tested. The data showed that most of the differentiated cells formed cell aggregates after the 30th day, with more cells accumulated to form spheroids. RT-PCR revealed the expression of the hepatic nuclear factor, albumin, cytokeratin 18 and 19, cytochrome P2B6, glucose-6-phospahtase and claudin. The functional assays also showed glycogen storage and omission of indicynine green. Our study demonstrated hBSCs are novel population that can differentiate into hepatocyte-like cells.