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Int J Med Sci 2022; 19(1):98-104. doi:10.7150/ijms.65343 This issue Cite

Research Paper

RNA isolation from micro-quantity of articular cartilage for quantitative gene expression by microarray analysis

Xiaowei Zhang, MD^1,2, Trevor J. McFarland, BS³, Kristina Vartanian, BS³, Yong Zhu, PhD⁴, Christina A. Harrington, PhD^3,5, Cong-Qiu Chu, MD, PhD^{1,2 ✉}

1. Division of Arthritis and Rheumatic Diseases, Oregon Health & Science University, Portland, Oregon 97239.
2. Section of Rheumatology, VA Portland Health Care System, Portland, Oregon 97239.
3. Gene Profiling Shared Resource, Oregon Health & Science University; Portland, Oregon 97239.
4. Vivoscript, Inc, P. O. Box 63025, Irvine, CA 92602.
5. Department of Molecular and Medical Genetics, Oregon Health & Science University; Portland, Oregon 97239.

✉ Corresponding author: chucedu

Abstract

Isolation of quality RNA from articular cartilage has been challenging due to low cellularity and the high abundance of extracellular matrix and proteoglycan proteins. Recently developed methods for isolation of high quality RNA from cartilage are more applicable to larger cartilage specimens typically weighing at least 25 mg. While these methods generate RNA suitable for analysis, they are less successful with smaller tissue inputs. For the study of small focal defect cartilage specimens an improved RNA extraction method is needed. Here we report a protocol for direct RNA isolation from less than 3 mg of wet weight rabbit articular cartilage for quantitative microarray gene profiling. This protocol is useful for identifying differentially expressed genes in chondrocytes following focal cartilage repair and can potentially be adopted for gene expression analysis of cartilage biopsy specimens from human joints.

Keywords: RNA isolation, articular cartilage, cartilage repair, microarray

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