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

Table 1 Example of the research structure in PubMed

The same search strategy was used in the other databases
PubMed	Human Breastmilk OR Human Milk OR
(Title/Abstract)	Breast Feeding OR Lactation
AND
PubMed	Stem Cells Or Progenitor Cells OR
(Title/Abstract)	Mother Cells
NOT
PubMed	Cancer
(Title/Abstract)

Table 2 Details of selected articles

Ref.	Purpose	Number of samples	Vol. of milk collected	Income cellcells/ mL	Markers - FC	Markers - IF	PCR	WB	Differentiation cell	Conclusion
Patki et al[12]	To study the multipotent differentiation capacity of the long-term MSC culture.	35	15 mL	2.5 x 106-3.0 x 106/mL	CD29, CD33, CD34, CD44, CD45, CD73, SCA-1.	Nestin, Vimentin, Smooth Muscle Actin and E-Cadherin.	Not done	Not done	Adipogenic, chondroge-nic, osteogenic lineages.	Mesenchy-mal stem cells isolated from human breast milk have cellular potency to differentiate into different cell types.
Hassiotou et al[13]	To examine regulators of self-renewal of hBSCs and their plasticity and potential to differentiate into cell types outside the mammary lineage.	> 70	5-200 mL	Not done	SSEA-4 e TRA-1-60 / TRA-1-81.	OCT4, SOX2 and Nanog.	TRA-1-60 / TRA-1-81, Oct4, Sox2, Klf4 and Nanog genes.	Oct4, Sox2 and Nanog.	Osteogenic, adipogenic, chondroge-nic, neuronal, pancreatic, hepatocyte, cardiomyo-cytes, mammary differentiation and teratoma.	Stem cells in breast milk contain pluripotency properties.
Hosseini et al[14]	To differentiate the breast milk-derived stem cells toward neural stem cells and then into the neurons and neuroglia.	Not present	5-200 mL	4.0 x 104/mL	Not done	Nestin, Nanog, β-tubulin III, O4, GFAP, Oct4, Sox2, CD44, CD105, CD106, CD133.	Not done	Not done	Neurosphe-res, neurons, oligodendro-cytes, and astrocytes.	The breast milk-derived stem cells showed the capability to be differentia-ted into neural cell lineages and their similarity to both embryonic and mesenchy-mal stem cells makes them a good candidate for cell therapy in neurodegenerative diseases.
Sani et al[15]	To examine the pluripotency of the human breast milk-derived cells.	5	5-50 mL	Not done	CD90 CD44, CD144, CD 271, CD123, CD133, CD73, CD 106, CD146, CD45, CD 105,CD34, SSEA-4, TRA-60–1, CD 15/SSEA-1.	Oct4, Sox2, Nanog and CK18.	SRY gene	Not done	Osteogenic and adipogenic lineages.	Most of the cells found are mesenchy-mal stem cells, but, cells similar to embryonic stem cells are also present.
Sani et al[16]	To examine the differentiation potential of the hBSCs into functional hepatocytes in vitro.	15	5-50 mL	Not done	Not done	CK18, CK19, albumin, alpha-fetoprotein.	AFP, CK19, CYP2B6, HNF4, G6P.	Not done	Hepatocyte lineage.	The hBSCs demonstrate the ability to express many key factors that are important in liver functions.

hBSC: Human breastmilk stem cell.

Table 3 CD and their functions

Marker	Function	Ref.
CD15/SSEA-1	Stage-specific embryonic antigen, also marker for primitive mesenchymal cells in human bone marrow.	Anjos-Afonso et al[17]
CD29	Expressed in myoepithelial cells including mammary gland cells.	Indumathi et al[5]
CD33	Present in immune cells of myeloid lineage.
CD34	Marker of hematopoietic stem cells, vascular endothelium and fibroblasts of some tissues.	Indumathi et al[5]
CD44	Marker of myoepithelial cells.	Indumathi et al[5]
CD45	Membrane protein present in lymphocytes and absent in erythrocytes and platelets. This protein is lost during the maturation of red cells in the bone marrow. Because it is present in lymphocytes, it is called Common Leukocyte Antigen.	Notta et al[18]
CD73	Marker expressed in mesenchymal stem cells, possessing enzymatic activity and catalytic dephosphorylation of adenosine monophosphate converting it into adenosine.	Indumathi et al[5]
CD90	Marker of mesenchymal stem cells.	Indumathi et al[5]
CD105	Marker of mesenchymal stem cells.	Indumathi et al[5]
CD106	Acts on the cell cycle of T and B lymphocytes, in addition to being involved in tissue repair.	Indumathi et al[5]
CD123	The IL-3 receptor subunit plays an important role in the growth and differentiation of hematopoietic progenitor cells.	Testa et al[19]
CD133	Marker of hematopoietic stem cells.	Indumathi et al[5]
CD144	Also known as VE-cadherin and cadherin-5, it is a calcium-dependent transmembrane cell adhesion molecule, located in the intercellular borders of endothelial cells, hematopoietic stem cells and perineural cells.	Dejana et al[20]
CD146	Mediated cell-cell interactions, cytoskeletal remodeling, angiogenesis, and migration of endothelial cells.	Sorrentino et al[21]
CD271	Transmembrane protein found in neuronal axons, Schwann cells, and perineural cells of peripheral nerves. In addition, it can be found in some epithelial, mesenchymal, and lymphoid tissues.	Lv et al[22]
SSEA-4	Marker of mesenchymal stem cells derived from the BM, acting mainly on cell proliferation.	Lv et al[22]
TRA-60-1	Related to pluripotency of human embryonic stem cells, and lost in cell differentiation.	Schopperle et al[23]
NESTIN	Multipotent, neural, medullary, pancreatic, and epithelial stem cell marker.	Toma et al[24]
VIMENTIN	Cytoskeletal intermediate filament protein, associated with the nuclear and plasma membrane, maintains the position of the nucleus and the mitotic spindle during the life of the cell, and is found mainly in mesenchymal stem cells.	Mendez et al[25]
SMA	Marker of myoepithelial cells.	Twigger et al[26]
E-CADHERIN	Calcium-dependent cell adhesion molecule, with important function in the formation and maintenance of normal tissue architecture, present in epithelial cells of the human mammary gland and epithelial marker of mesenchymal transition in its first passages.	Klopp et al[27]
NANOG	Pluripotency marker.	Riekstina et al[28]
β-TUBULIN III	Considered an early neuronal marker because the tubulin protein is the main constituent of the microtubules, which are tubular structures that make up the cytoskeleton and they are involved in the transport of organelles and the elongation of axons and dendrites.	Kapitein et al[29]
O4	Specific marker of gangliosides expressed by pre-oligodendrocytes and premyelinated oligodendrocytes.	Girolamo et al[30]
GFAP	Glial fibrillary acidic protein forms subunits of the intermediate filaments of the cellular cytoskeleton, being present in the cytoplasm of the astrocytes.	Hol et al[31]
OCT4	Pluripotency marker.	Riekstina et al[28]
SOX2	Controls the undifferentiated and pluripotential state of the CTEs.	Young et al[32]
CK18	Expressed in the luminal cells of the breast, which synthesize proteins from breast milk.	Thomas et al[33]
KLF 4	Kruppel like factor 4 is a transcription factor related to pluripotency and cell proliferation potential.	Li et al[34]
SCA-1	The stem cell antigen 1 is found in stem cells.	Holmes et al[35]