Cell and Tissue Culture Course - SCBZ 432 PDF

Cell and Tissue Culture Course SCBZ 432 By Dr. Ghadeer Alrefaei 1 Cell and Tissue Culture Course Reference References available in this links :...

Cell and Tissue Culture Course SCBZ 432 By Dr. Ghadeer Alrefaei 1 Cell and Tissue Culture Course Reference References available in this links : https://www.amazon.com/Principles-Practice-Animal- Al-karim Saleh,Al-Qudsi Tissue-Culture/dp/8173717192 Sudha Gangal(2010). Principles and Practice of Fatma, Khorshid Faten Animal Tissue Culture.2nd (2010).Introduction to https://www.researchgate.net/publication/261252418_Intr edition nature, stem, cancer cell oduction_to_nature_stem_cancer_cell_culture_in_arabic culture. 1st edition 2 Lecture 2 : Types of cell culture in vitro objectives: 1. Identify the are three major types of cell culture 2. Knowing the Characteristics of Primary cell culture 3. Understand the difference between finite and Continuous cell lines 4. Transformation VS Transfection in Continuous cell lines. 3 In vitro culture (maintain and/or proliferation) of cells, tissues or organs. 4  Three major categories of tissue culture Primary cell culture Secondary cell culture or subculture Cell line  There are three major types of cell culture in vitro 5 How can cell line be derived? 6   Three major categories of tissue culture Primary culture term refers to that stage of the Primary cell culture culture after the isolation of cells from tissue, where the cells are proliferated under Secondary cell culture or appropriate conditions until they occupy all of subculture the available substrate. In the other words, it's a stage of the culture after the isolation of the Cell line cells but prior to the first subculture after which it becomes a cell line.  There are three major types of cell culture in vitro 7  Secondary cell culture or subculture  The growth of cells in culture proceeds from the lag phase following seeding to the log phase, where the cells proliferate exponentially.  When the cells in adherent cultures occupy all the available substrate and have no room left for expansion, or when the cells in suspension cultures exceed the capacity of the medium to support further growth, cell proliferation is greatly reduced or ceases entirely.  To keep them at an optimal density for continued growth and to stimulate further proliferation, the culture has to divide and fresh medium supplied. 8 Secondary cell culture  Three major or of categories subculture tissue culture  Primary Cell culture cannot remain viable for a long time because the cell utilize all nutrients of the medium, therefore, sub-culturing is needed.  Secondary culture or Secondary cell culture refers to the culture formed after sub-culturing of primary cell culture.  Sub-culturing also referred to as passaging, is the removal of the medium and transfer of cells from a previous culture into a fresh growth medium, a procedure that enables the further propagation of the cell line or cell strain. 9  Secondary cell culture or subculture Derived from primary cell culture and isolated by selection or cloning. Becoming a more homogeneous cell population. Finite life span in vitro. Retain differentiated phenotype. Mainly anchorage-dependent. Exhibit contact inhibition. 10  Cell line  A cell line is a permanently established cell culture that will proliferate indefinitely given appropriate fresh medium and space.  A cell culture developed from a single cell and therefore cell line consists of cells with a uniform genetic make-up.  A cell line is a cell population derived from the primary cell line.  Finite cell line  Continuous cell lines  Finite indicates a limited life span and continuous represents an unlimited life span, and have two type ( Normal cells or Transformed Cells ) 11  There are two types of continuous cultures Cell lines (finite cell line) Continuous cell lines Finite and Continuous cell lines Normal cells usually divide only a limited number of times before losing their ability to proliferate, which is a genetically determined event known as senescence; these cell lines are known as finite. However, some cell lines become immortal through a process called transformation, which can occur spontaneously or can be chemically or virally induced. When a finite cell line undergoes transformation and acquires the ability to divide indefinitely, it becomes a continuous cell line 12 13  Cell Culture Terminology  Cell culture - Maintenance of dissociated cells in culture  Tissue culture - Maintenance of tissue explants in culture  Cell line - A culture that is sub-cultured beyond the initial primary culture phase  Finite cell line - A cell line with a limited lifespan that eventually undergoes senescence  Continuous cell line - A cell line that is essentially immortal and continues indefinitely  Primary culture - The initial culture derived from in vivo material  Clone - The progeny isolated from a single cell  Immortalization - Enabling of cells to extend their life in culture  Contact inhibition - is a growth mechanism. In most cases when two cells collide they attempt to move in a different direction to avoid future collisions.  Substrate- The matrix on which a culture is grown  Passage -Subculture of cells from one container to another 14  Video https://youtu.be/9BvTFowr0rI 15 The end 16