Good manufacturing practice-compliant isolation and culture of human bone marrow mesenchymal stem ce

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Good manufacturing practice-compliant isolation and culture of human bone marrow mesenchymal stem cells

Phuc Van Pham, Nhan Lu-Chinh Phan, Dung Minh Le, Phuong Thi-Bich Le, Tung Dang-Xuan Tran, Ngoc Kim Phan

Abstract

Bone marrow mesenchymal stem cells (BM-MSCs) are multipotent stem cells that can differentiate into some kinds of mesenchymal cells such as osteoblasts, chondroblasts and adipocytes. These cells were discovered for a long time and considered as the first discovered source of MSCs. BM-MSC transplantation was used to treat some diseases such as bone disease, myocardial infarction, stroke, diabetes mellitus… This study aimed to provide a new method to in vitro primarily culture and secondary culture of BM-MSCs that were compliant with good manufacturing practice for clinical applications. Bone marrow was aspirated from the hip bone using special needles and syringes. Mononuclear cells (MNCs) were isolated from bone marrow by BM-MSC extraction kit. These MNCs were cultured in the commercial medium – MSC-Cult supplemented with 10% activated platelet rich plasma (aPRP) to obtain BM-MSCs. The results showed that 100% samples of bone marrow can be successfully cultured to produce BM-MSCs. Obtained cells exhibited the MSC phenotypes, and maintained the stable and normal karyotype up to 10th passages, and failed to cause tumor in the mouse models. This research provided an advanced protocol for clinical applications of BM-MSCs.

Keywords

Bone marrow mesenchymal stem cells; Clinical application; GMP; Mesenchymal stem cells; Stem cell culture

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DOI: http://dx.doi.org/10.15419/psc.v1i1.48