Question

a) Characterization of cell lines are important phenomena for cell culture experiments. Discuss briefly why we need to carefully characterize cell lines.

, b) What is the simplest cell characterization technique? Based on this characterization you observed one cell line fibroblastic and the other one epithelial. Please briefly explain their morphologies.

c) What would be the consequences of working with not properly characterized cell lines?

d) How would you sterilize liquid based materials that is used in cell culture experiments?

Answer 1

a - Cell line characterization is critical to the control of biological product manufacturing. The main objective of the testing is to confirm the identity, purity, and suitability of the cell substrate for manufacturing use. The specific strategy of testing to characterize a cell bank varies based on several factors including, but not limited to, the source of the cell line itself, its growth requirements, and cultivation history.

b - Morphological characterization is the most simleast form of characterization of cells plays an important role in clinical diagnosis .
For example, tumorous cells are crowded and enlarged compared to normal cells . Microscopy is routinely used to examine the cellular morphology, but it is not applicable to in vivo tissue. It has been shown that multispectral diffuse optical tomography (MS-DOT) is a promising modality for imaging of cellular morphology since it is non-invasive and can be used to in vivo characterize cells located deep in tissue

1. Fibroblastic (or fibroblast-like) cells are bipolar or multipolar, have elongated shapes, and grow attached to a substrate.
2. Epithelial-like cells are polygonal in shape with more regular dimensions, and grow attached to a substrate in discrete patches .

c - Cell culture contamination’ is the most harmful effect of using unsterlized cell lines that typically conjures thoughts of bacteria amongst cells and cloudy media, unwanted invaders in the culture flask can take many forms. Viral and chemical contaminants can also have considerable consequences for culture health, and ensuring that cell lines are not cross-contaminated is critical to reproducibility of results.

d -Liqid based material if contain cell lines can not be sterlized by dry or wet sterlization method so fiteration is used -

FILTRATION

Filters work by passing the solution through a filter with a pore diameter that is too small for microbes to pass through.

Filters can be scintered glass funnels made from heat-fused glass particles or, more commonly these days, membrane filters made from cellulose esters. For removal of bacteria, filters with an average pore diameter of 0.2um is normally used.

But remember, viruses and phage can pass through these filters so filtration is not a adequate for them .
Distilation process are also used for sterlization .