Figure 2 - uploaded by Satish Patki
Content may be subject to copyright.
Context in source publication
Context 1
... the study of the mesenchymal stem cells, the protocol of culture as on different days established in our earlier original study [1] was used. Breifly, the cell pellet was seeded in a 35 mm tissue culture dish (BD falcon) using Dulbeccos modified eagle medium (DMEM), containing 10% heat inactivated human umbilical cord blood serum (HUCBS), supplemented with penicillin (100 units/ml) and streptomycin (100 μg/ml). The dish was incubated at 37°C under 5% CO 2 and 95% humidity. Medium was changed every 48 hours. The cells were observed under phase contrast microscope at 400 x magnification. At 80% confluency, cells were passaged using trypsine ethylene diamine tetra acetic acid (EDTA). The cells from early (2 to 6) passages were used for the characterization studies. The cells were dislodged using 0.05% trypsin, 0.02% EDTA in PBS and resuspended in DMEM. The cells were fixed in chilled 70% ethanol and incubated in mouse antihuman fluorescein isothiocynate (FITC) Phycoerythrine (PE) conjugated antibodies against CD33, CD34, CD44, CD45, CD73, CD90 and CD117 for one hour on ice. The cells were acquired using a flow cytometer laser 488NM (Becton Dickinson, New Jersey, NJ, USA) and data were analysed BD Cellequest pro software. Table 1 shows the clinical data of the study subjects. Table 2 shows the average cell counts and the percentage of the leucocytes and the epithelial cells observed on day 0.5 and 7. The total cell counts and the percentage of leucocytes are significantly higher on day 5 of lactation as compared to day 0. (P<0.01) However, the percentage of the epithelial cells is found to be steady throughout the week. Figure 1 shows the cluster of epithelial cells while Figure 2 shows the various types of leucocytes observed on day 5 of lactation. Figure 3 shows the mesenchymal stem cells (MSCS) adherent to the petridish with fibroblast like appearance. The adherent cell population is a mixture of epithelial cells from mammary gland alveoli and mesenchymal stem cells (MSCS). However, during the second week, the cells in the culture flask resembled a typical slender fibroblast – like cell phenotype. This could be because of epithelial to mesenchymal transition which is commonly observed in development and regeneration. The immunofluorescence study for specific cell surface Page 2 of 3 marker clearly indicated that isolated human breast milk cells at passage four expressed mesenchymal stem cells (MSCS) markers, namely CD44, CD29, SCA1. These cells were found to be negative for CD33, CD34, CD45, CD73 by fluorescence activated cells sorting analysis confirming their identity as mesenchymal stem cells (MSCS). Composition of the human milk is indeed a topic of research over the years. Apart from the nutritional value, it provides many ...
Similar publications
Citations
... Accordingly, the human milk was defined as a dynamic fluid, with all necessary nutrients and growth factors useful for the optimal development of the newborn, but also containing huge amounts of maternal live cells. 7 The mix of all constituents was considered fundamental for the optimal growth of the neonate. In following studies, the lactating mammary gland revealed its intimate nature: to represent a source of a heterogeneous population of breast-derived cells, including exfoliated luminal epithelial cells, immune cells, mesenchymal stem cells and embryonic stem cells, giving rise to a peculiar stem/progenitor cell pool. ...
Recent findings of stem/progenitor cells in maternal milk and their ability to cross the intestinal barrier of lactating newborns and integrate into neonatal organs to promote optimal child development present a new challenge in perinatal medicine. These findings emphasize the need for all mothers to breastfeed their babies for a long time. According to recent research, breastfeeding protects the lactating newborn from multiple infectious agents that can cause severe and fatal early infancy diseases. The second benefit is that maternal stem cells accelerate the development of several organs, including the brain, protecting lactating infants from severe childhood and adult diseases. The success and diffusion of exclusive breastfeeding, especially in low-resource settings, depends on mothers' knowledge of the many benefits for their child, including recent discoveries on breastfeeding's powerful benefits. Every mother may need simple booklets to learn about the unique benefits of maternal breastfeeding, including the nutrients and multiple cell types that protect the newborn from infections and accelerate neonatal organ development. Social media should also be encouraged to spread news about breastfeeding and maternal stem cells' impact on lactating infants' health. Health belief model interventions may boost breastfeeding. In conclusion, the discovery of massive amounts of cells in maternal milk and the identification of stem/progenitors with previously unknown potential in newborn development after birth should be considered a new valuable tool for exclusive breastfeeding advocates. Data here suggests that every action to spread this message and educate mothers and families about breastfeeding's irreplaceable role is mandatory.
... They describe normal cells found in human milk and they thought that milk harbors epithelial cells and immune cells. The immune cells in the human milk consist of macrophages (large lipidladen macrophages), neutrophils, and lymphocytes of which the majority are T cells, also this study's results go with what stated with Hassiotou et al., have also observed that 70% of the total human milk cells in the first two postpartum weeks consist of immune cells [22]. In this study NAF was produced twenty-two percent (22/100) this result is different from what was reported by the proctor at that. ...
Background: Breast cancer is common in Sudan and most patients are detected at later stages of the disease due to the lack of awareness and absence of early screening programs. The majority of breast cancers originate in the epithelium lining the milk ducts. It is believed that most breast canceras are slow growing and progress from precancerous cells, which have cellular and nuclear changes that can be identified microscopically. Aim: To cytological screening of breast cancer in breast fluid. Methods: This is a cross-sectional feasible study conducted in Shendi town, 200 samples of different breast fluids from asymptomatic women. (100 nipple aspiration fluid, 50 milk smear, 50 postpartum milk) have been collected and screened cytologically. Stained by pap stain. Results: After cytology screening of breast fluid in, milk 30/50(60%) of women were non-cell secretors and 20/50 (40%) of women were cell secretors. Cells that appeared among the secreting group were a few epithelial cells and immune cells. In postpartum milk all women 50/50 (100%) were cell secretors, abnormal cytomorphological changes were in 15/50 (30%) of them, and high secretion of immune cells 50/50 (100%) which found to statistically of significant value (0.000). NAF was not produced in 78/100. Among women who produced NAF22/100, women were produced NAF 6/22 (27.3%) were not cell secretors (category 0), benign nonhyperplasic ductal epithelial cells (category 1) 6/22(27.3%), benign hyperplasic ductal epithelial cells (Category II) 5/22(22.7%), atypical ductal epithelial cell (category III) 3/22 (13.7%) atypia (Category IV) 2/22(9%). cytological atypia 21/25 (84%) appears among women more than 30 years old and there was a strong statistical of significant value P. value =(0.000). Also, risk factors (family history, contraceptive intake, and HPV) statistically have a significant correlation with cytological atypia among study group P. value = (0.036). In this study HPV infection, cytomorphological change (koilocyte) was detected in breast fluid 4/200 (2%). Conclusion: cytology of the breast is a simple, safe, rapid test that is acceptable to patients and showed the ability to detect benign and pre-neoplastic ductal epithelial cells from asymptomatic volunteers.
... Lisozim merupakan suatu enzim yang dapat menghancurkan dinding bakteri gram positif dan gram negatif (Golinelli et al., 2014). Beberapa penelitian terhadap enzim menunjukkan bahwa proses freezing dapat menyebabkan protein kehilangan sebagian struktur tersier, agregasi protein, destabilisasi struktur sehingga kehilangan fungsi katalitik, serta denaturasi protein (Bortolin et al., 2017;Gabellieri dan Strambini, 2003;Franks, 1995) Fraksi pelet pada ASIP mengandung banyak sel imun yaitu makrofag, netrofil dan limfosit (Patki et al., 2014). Makrofag adalah leukosit dengan jumlah paling banyak sebesar 40 -50% dari total leukosit pada kolostrum ASI diikuti oleh neutrophil (40 -50%), sedangkan limfosit sekitar 5 -10% dari total leukosit . ...
Sumber nutrisi utama untuk pertumbuhan dan proteksi bayi yang mengandung makronutrisi, mikronutrisi, dan senyawa bioaktif adalah air susu ibu (ASI). Kegagalan menyusui dialami lebih banyak oleh ibu yang bekerja. Saat ini, pemberian ASI perah beku merupakan solusi alternatif tetapi kemampuan senyawa imunologis ASI perah beku dalam menghambat bakteri penyebab diare mungkin mengalami penurunan. Tujuan penelitian ini adalah membandingkan dan mengetahui kemampuan fraksi ASI segar dan beku dalam menghambat pertumbuhan bakteri Enterotoksigenik Escherichia coli (ETEC). Penelitian ini menggunakan quasi-experimental design dengan tujuh sampel ASI dengan kondisi empat belas hari pasca melahirkan dan disentrifugasi untuk mendapatkan fraksi supernatan dan pelet. Hasil penelitian ini adalah ada penurunan yang signifikan pada whole milk, supernatan, dan pelet ASI segar maupun beku dalam menghambat pertumbuhan ETEC. Fraksi supernatan pada ASI segar dan beku memiliki kemampuan lebih tinggi dalam menghambat bakteri dibandingkan dengan whole milk dan fraksi pelet.
... It contains a plethora of proteins, fat-soluble vitamins, minerals, immunoglobulins, various cellular components and growth factors. The detailed cellular composition of human colostrum is well defined 5,6 . Soon after parturition, the secretion of colostrum lasts for approximately two weeks, after which mature milk formation starts, which lasts till involution. ...
Recently, breastmilk has been observed to be an ideal source of stem cells. Here,
different cellular constituents that are present in human breastmilk, identified by
cytological staining procedures. Smears were prepared from breastmilk and fixed
with ether and 95% ethyl alcohol (1:1) followed by staining with Giemsa,
Hematoxylin and Eosin. Different cell types were observed. These were identified by
their unique morphological patterns and archetypal geometry of the nucleus.
Monocytes have metaphorically bean/ kidney-shaped or horse-shoe shaped or heart
shaped nucleus whereas lymphocytes have large nucleus. Macrophages display
irregular shape. Plasma cells have small, dense, eccentric nucleus, voluminous
cytoplasm. Neutrophils, basophils and eosinophils were recognized by their lobulated
nucleus. Lactocytes are characterized by exclusive lipid inclusions. There was a
predominant occurrence of vacuolar space resembling morphologically to alveolar
and lobular cells of functional mammary gland of human. The expression of the
following cell types in mature human milk concludes that they have both
hematopoietic and mammary origin. This study further illustrates the possibilities of
breastmilk to consider as a unique cellular model that can be used to study breast
pathology and biology of mammary glands
... It contains a plethora of proteins, fat-soluble vitamins, minerals, immunoglobulins, various cellular components and growth factors. The detailed cellular composition of human colostrum is well defined 5,6 . Soon after parturition, the secretion of colostrum lasts for approximately two weeks, after which mature milk formation starts, which lasts till involution. ...
Keywords: breastmilk, stem cells, cellular constituents, staining, cell types, hematoxylin and eosin, diagnostic cytology. Recently, breastmilk has been observed to be an ideal source of stem cells. Here, different cellular constituents that are present in human breastmilk, identified by cytological staining procedures. Smears were prepared from breastmilk and fixed with ether and 95% ethyl alcohol (1:1) followed by staining with Giemsa, Hematoxylin and Eosin. Different cell types were observed. These were identified by their unique morphological patterns and archetypal geometry of the nucleus. Monocytes have metaphorically bean/ kidney-shaped or horseshoe shaped or heart shaped nucleus whereas lymphocytes have large nucleus. Macrophages display irregular shape. Plasma cells have small, dense, eccentric nucleus, voluminous cytoplasm. Neutrophils, basophils and eosinophils were recognized by their lobulated nucleus. Lactocytes are characterized by exclusive lipid inclusions. There was a predominant occurrence of vacuolar space resembling morphologically to alveolar and lobular cells of functional mammary gland of human. The expression of the following cell types in mature human milk concludes that they have both hematopoietic and mammary origin. This study further illustrates the possibilities of breastmilk to consider as a unique cellular model that can be used to study breast pathology and biology of mammary glands.
Objectives
Breast milk is the best baby food because it contains various nutrients and important factors for the baby's immune system, including leukocytes. This study aimed to determine the effects on morphology, number of cells and breast milk leukocytes count of various ways of storing breast milk based on different temperatures and storage durations.
Methods
This study was conducted at the Biochemistry Laboratory, Integrated Laboratory and the Histology Laboratory, Universitas Indonesia, Jakarta, Indonesia from September 2022 to February 2023. Transitional breast milk samples from 7 breastfeeding mothers were utilised in the study. A total of 50 mL was divided into 4 tubes of 12.5 mL each and treated based on temperature, storage time and method of thawing frozen breast milk based on the Centers for Disease Control and Prevention's (CDC) recommendations for breast milk storage. The breast milk cells were isolated to calculate the cell number and leukocyte population. Subsequently, the breast milk cells were stained with haematoxylin and eosin to analyse the number and morphology of leukocytes.
Results
The findings showed a significant decrease in the breast milk’s total number and population and changes in the morphology of breast milk leukocytes after storage.
Conclusion
This study indicates that CDC storage recommendations do not affect the quantity of the CD45+ leukocyte population; however, there is a decrease in the total number of leukocytes and alterations in their microscopic morphology. Thus, additional research is recommended to determine whether these modifications influence the function of the breast milk cells.
Cellular components of breast milk, along with the nutritional component, provides the benefit of growth, development and protection to the newborns. Breast milk comprises of epithelial cells, phagocytes, mammary stem cells, hematopoietic stem cells, mesenchymal stem cell and other progenitor cells. The role of milk derived stem cells in the newborn is yet unclear. However, studies on animal models have shown the presence of maternal stem cells in the organs of infants, suggesting their possible role in the adult life. Furthermore, the source of stem cells in human milk and the regulatory pathways governing the functions of these stem cells in infants need to be explored. Thus, it is of great interest to isolate and characterize the stem cells derived from breast milk and study their possible implications in the field of regenerative medicine.KeywordsMammary stem cellsHematopoietic stem cellsMesenchymal stem cellsProgenitor cellsMaternal stem cellsOrgans of infantsSource of stem cellsRegulatory pathways
