Expansion and characterization of cells from surgically removed intervertebral disc fragments in xenogen-free medium
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Expansion and characterization of cells from surgically removed intervertebral disc fragments in xenogen-free medium SIMRAN MUJAWAR1, KRUTTIKA IYENGAR2, SUNIL NADKARNI1* and RITA MULHERKAR2* 1
Smartha Krupa Life Sciences, BKL Walawalkar Rural Hospital, Ratnagiri, India 2
Moving Academy of Medicine and Biomedicine, Pune 411 016, India
*Corresponding authors (Emails, [email protected]; [email protected]) MS received 2 May 2020; accepted 26 August 2020 Low back pain due to degeneration of intervertebral disc (IVD) is a major health problem resulting in significant disability as well as adding to the economic burden. Discectomy is a very common procedure done worldwide to relieve this pain. At present all the surgically removed disc tissue is mostly discarded. However, there are reports that state that progenitor cells in the IVD can be grown ex vivo and have the potential to be used for IVD repair and regeneration. We report here that viable cells can be harvested from surgically removed, herniated disc tissue and can be potentially used in cell based therapy. Further, we have successfully replaced xenogenic supplements such as foetal bovine serum with either autologous serum or human platelet lysate for culturing IVD cells from patient’s surgically removed disc tissue, without loss of any cell characteristics, including cell surface markers, growth factor secretion in the conditioned medium and osteogenic and chondrogenic differentiation potential in vitro. The present work will not only contribute to overcoming some of the major barriers in carrying out human clinical trials, but also provide a cheap, alternate source of proteins and growth factors for growing IVD cells ex vivo for therapy. Keywords. Autologous serum; cell-based therapy; intervertebral disc; mesenchymal stem cell; non-xenogenic supplements; platelet lysate
1. Introduction Herniated intervertebral (IVD) disc tissue is the most common cause of back and leg pain adding to the economic burden (Vos et al. 2012). The vertebral column is made of segments with IVD which is a fibrocartilaginous tissue situated between two adjacent vertebrae and contributes to motion, weight-bearing and flexibility of the spine. The IVD itself consists of a hydrophilic, gelatinous core, the nucleus pulposus (NP) which is surrounded by several layers of radially arranged collagen fibres forming the annulus fibrosus (AF) (Sakai and Schol 2017). A tear in the AF results in protrusion of disc tissue. The loss of disc tissue sets up a degenerative cascade in the motion segment. Due to disc desiccation and gradual loss of disc height there is narrowing of the space available for the nervous tissue causing limitations in walking and exercise http://www.ias.ac.in/jbiosci
tolerance hampering the individual’s effort to maintain good health through exercise. The degenerated disc protruding out of AF is often surgically removed to relieve pain. Considerable number of disc tissue fragments which contain reg
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