Technique / Cell Biology / Cell culture / Primary cell culture

Subculture procedure for monolayer cells new protocol
This protocol was taken from "Primary Culture of Human Proximal Renal Tubular Epithelial Cells" by Paul A. Glynne. From: Methods in Molecular Medicine, Vol.36: Septic Shock. Humana Press Inc., Totowa, NJ. (The Kidney Research Group, Royal British Hospital, Aus ...

Protocol for primary culture of human umbilical vein endothelial cells (HUVEC) new protocol
A detailed protocol for primary cultures of HUVECs including buffer and reagent preparation, protocol and image. (PDF file, Professor Bruno Baudin) ...

Discontinuous density Percoll gradient separation and primary culture of bovine chromaffin cell new protocol
This protocol uses discontinuous density Percoll gradient protocol to purify bovine chromaffin cells from 3 bovine adrenal glands for primary culture of chromaffin cells. ...

Mouse Adrenal Chromaffin Cell Isolation Protocol Video media
Mouse Adrenal Chromaffin Cell Isolation online video demonstration. Aaron Kolski-Andreaco1, Haijiang Cai2, D. Spencer Currle3, K. George Chandy1, Robert H. Chow2 1Department of Physiology and Biophysics, University of California, Irvine, 2Department of Ph ...

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Establishment of fibroblast cultures.
(Curr Protoc Cell Biol. 2001 May;Chapter 2:Unit 2.1.)
Takashima A.

University of Texas Southwest Medical Center, Dallas, Texas, USA.

Fibroblasts are fairly easily isolated from a variety of tissues, in this case
skin. The cells are fast growing and can be rapidly expanded from small samples.
They are suitable for behavioral, functional, biochemical, and genomic studies.
Skin fibroblast cultures are started from explants in which the epidermis is
enzymatically removed to prevent contamination of the fibroblasts with epidermal
cells. Alternatively, the dermis can be enzymatically dissociated to provide a
suspension of fibroblasts.

Preparation and culture of human lymphocytes.
(Curr Protoc Cell Biol. 2001 May;Chapter 2:Unit 2.2.)
Biddison WE.

National Institute of Neurological Disorders and Stroke/NIH, Bethesda, Maryland,
USA.

This unit presents protocols for preparation of lymphocytes from peripheral
(whole) blood or leucopherisis samples, first by differential centrifugation in
Ficoll-Hypaque to separate them from erythrocytes and granulocytes. Repeated
centrifugation removes the plasma and platelets. Monocyte/macrophage cells and
dendritic cells can be purified from the T and B cell population by exploiting
their adhesion to serum-coated plates in the presence of recombinant IL-3.
Different subpopulations of lymphocytes can be isolated by specific antibodies
bound to magnetic beads.

Preparation of endothelial cells
(Curr Protoc Cell Biol. 2001 May;Chapter 2:Unit 2.3)
Kleinman HK, Cid MC.

National Institute of Dental Research/NIH, Bethesda, Maryland, USA.

Endothelial cells, which line blood vessels, can be prepared from a variety of tissues. They are frequently prepared from the umbilical vein, which is relatively easy to obtain. The procedure is clearly described and provides a large population of highly purified endothelial cells. There are also methods for obtaining endothelial cells from other tissues such as fat, skin, and mucosa. These methods require special care and generate smaller populations of cells.