Tissue Culture & Animal Biotechnology: Splitting Ratio Quiz

Key Figures in Cell Culture

• George Harrison is often regarded as the father of cell culture.
• In 1907, Harrison cultivated frog nerve cells and observed their growth in vitro using the 'hanging drop' method.
• Alexis Carrel was the first to demonstrate the survival of cells isolated from blood and connective tissue in serum and plasma.
• Carrel also formulated the first liquid media consisting of sea water, serum, embryo extract, salts, and peptones.

Advances in Tissue Culture

• Carrel aimed to improve animal culture utilizing chick embryo tissue.
• In 1912, he reported the growth of chick tissue under sterile conditions by using serum-based media.

Cell Behavior in Tissue Culture

• Fibroblasts are generally the first type of cell to migrate in tissue culture.
• Cells that do not survive the dispersal method in tissue disaggregation often die and are not retained in culture.
• In primary culture, the cell population with the ability to proliferate most rapidly will dominate and become more prevalent.

Cell Culture Techniques

• Subculture (or passage) refers to transferring cells from one culture vessel to another to create new cultures.
• Cell lines with a limited lifespan are termed finite cell lines and undergo a limited number of cell generations.
• Continuous cell lines are typically generated through transformation or immortalization methods.

Understanding Split Ratios

• The split ratio in tissue culture indicates how the culture is distributed during subculture.
• A split ratio of 1:2 means that one part of the culture is transferred, while two parts remain in the original vessel.
• Tracking the number of elapsed doublings contributes to understanding growth rates and optimizing culture conditions.

Passage Number and Cell Differentiation

• Passage number represents the number of times a cell culture has been subcultured or transferred.
• Cell differentiation refers to the process by which cells become specialized in structure and function.
• Splitting the culture at specific ratios is important to maintain optimal density and health of cells.

Role of Hormones and Factors

• Retinoic acid is crucial in embryonic development, influencing differentiation pathways.
• Melanocyte-stimulating hormone (MSH) is categorized as a peptide hormone.
• Inducing agents, such as DMSO, can affect differentiation by altering signaling pathways.
• Adenylate cyclase is involved in differentiation processes by producing cyclic AMP, a secondary messenger.

Important Factors and Cryopreservation

• Keratinocyte differentiation is significantly influenced by growth factors in their environment.
• Vitamin D functions as a steroid hormone, important for calcium metabolism and cellular functions.
• The main purpose of cryopreservation is to preserve cells or tissues for future use.
• A common application of cryopreservation is the storage of embryos for in vitro fertilization (IVF).

Cryopreservation in IVF

• Cryopreservation enables couples undergoing IVF to store healthy embryos for future transfer.
• During frozen embryo transfer, previously cryopreserved embryos are thawed and implanted into the uterus.
• False statements about cryopreservation may include misconceptions regarding its success or safety.
• Potential risks associated with frozen embryo transfer include the possibility of embryo viability being affected upon thawing.

Test your knowledge on the concept of splitting ratio in tissue culture and animal biotechnology. Understand how to calculate the ratio of cells transferred to a new vessel compared to the total number of cells in the original culture.