Amniotic Epithelial cells

Available formats:≥ 2.5 x 106/ ml. cryopreserved in suitable medium

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General Description Specifications Certificate Of Analysis Technical Library

Normal Human Amniotic Epithelial Cells are present in the outer lining of the umbilical cord and are isolated using suitable explant techniques. These cells are predominantly found in the amniotic membrane, the cord’s outer connective tissue, primarily responsible for cushioning and nourishment of the developing fetus. The anchorage-dependent cell displays characteristic fibroblasts like morphology, similar to that of mesenchymal stem cells. However, these cells are easily distinguished from the MSCs due to the presence of distinct surface markers and intracellular proteins.At Kosheeka, we assure that our proliferating epithelial cells can be expanded for up to 15 population doubling. However, optimum culture conditions and expert handling play a crucial role in replicating the same at your facility. Rigorous quality control procedures are in place to certify that each batch of epithelial cells is negative for mycoplasma, bacteria and fungi. In addition to the same, we evaluate these cells for the presence of surface markers like CD46 and E-Cadherin as positive markers and CD105 as negative markers. We also examine the presence of intracellular proteins like cytokeratin and collagen I. However, this is not done for every batch. Upon specific requests assessments for these intracellular proteins and infectious markers like HIV, HCV etc. can be conducted.In order to ensure complete transparency and confidence in our products we share Certificate of Analysis with each shipment; which provides comprehensive quality control information to our customers.

Product Category:	Homo Sapiens, Human
Product Type:	Adherent, anchorage dependent population
Drived From:	Human Umbilical Cord Blood Cell
Cell Morphology:	Round, light emiting cells, with variable morphology
Culture and Growth Properties:	Cell doubling time of approximately 50-55 hrs.
Passage No:	As requested,
Mycoplasma:	Detected on demand
Hepatitis B:	Negative
Hepatitis C:	Negative
HIV-1:	Negative
HBsAg/HBcAg:	Negative
Aerobic & Anaerobic Bacteria:	Negative after 7 days of culture
Intracellular Markers:	Collagen I, Cytokeratin
Positive for :	CD 46, E-Cadherin
Negative for:	CD 105

Epithelial cells isolated from different locations of the body typically express specific expression profiles of surface markers and in vitro characteristics. Characterization commonly involves assessing cell morphology, cytokeratin expression, and growth patterns. Although these features are widespread, variations in expression profile based on cell’s origin and methodology used.

Certain epithelial cells, such as vaginal and mammary epithelial cells require specific substrates for cell adhesion and proliferation. On the other hand, epithelial cells derived from the amniotic membrane do not rely on any particular substrate for isolation, expansion and growth of epithelial cells.

In fact, the amniotic membrane is known as the richest source of epithelial cells, facilitating easy isolation, rapid doubling time and increased expansion capacity. Interestingly, initial passage cells are often a heterogeneous mix comprising both stem cells and fibroblasts, whereas later passages yield a purer population of epithelial cells.

Observations indicate that these cells can be expanded up to 10 passages without a significant impact on their doubling time. However, in the subsequent passages, these epithelial cells tend to acquire an elongated morphology, resulting in a substantial reduction in the cell count per flask.

Expansion of these cells do not require any cocktail of growth factors. Instead, a simple mixture of DMEM/F12 along with nutrient additives and 10% fetal bovine serum is enough for further expansion.

The expression of pancytokeratin is checked with the help of immunohistochemistry, which is a characteristic marker for the presence of epithelial cells. To differentiate between fibroblasts, connective tissues, and immune cells; certain specific surface markers like CD105, vimentin for fibroblasts and CD45/CD18 for leukocytes are employed. During the examination, epithelium and lamina propria (LP) are manually outlined and separately examined.

We estimated the cell doubling time from cells of multiple passages, such as passage number 3,6 and 9. While estimating the cell doubling time, cells were extracted and seeded in 24 well plates at a density of 10,000 cells per well.Subsequently cells were calculated from each well in triplicates at the same time daily for seven consecutive days, and the mean value was calculated using constructing growth curves. The population doubling time (PDT) was determined using the formula PDT= (T-T0) Ig2/(IgNt-Ig0)Where T0 represents the start time of the culture T represents the termination time of the culture N0 signifies the initial seeding cell count Nt indicates the final cell count

The cells are cryopreserved using a solution comprising 90% fetal bovine serum (FBS) and 10% dimethyl sulfoxide (DMSO). The cryopreservation process involves freezing the cells in a controlled rate freezer, gradually reducing the temperature to as low as -196 degree in liquid nitrogen. Once frozen the cells are stored for the estimated duration, maintaining their viability and preserving them, until further use.

Cells are shipped in liquid nitrogen or in dry ice in cryopreserved form. We can also ship cells in cultured form, maintaining their metabolic activity, and are ready for immediate use upon arrival.