Evaluation of different protocols for culturing mesenchymal stem cells derived from murine bone marrow

Pissarra, Mariana Ferreira; Torello, Cristiane Okuda; Saad, Sara Teresinha Olalla; Lazarini, Mariana

doi:10.1016/j.htct.2021.02.005

Cited by 1 publication

(1 citation statement)

References 27 publications

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“…To date, most of the MSC isolation methods have been based on the property of these cells to attach on the plasticware used for culturing [5,6]; however, in the case of mouse MSCs, this approach results in contamination with cells of the hematopoietic lineage [7,8]. To overcome this issue, several culture methods such as frequent medium exchange and low-density culture have been proposed, but these have certain limitations [9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Isolation of High Purity Mouse Mesenchymal Stem Cells through Depleting Macrophages Using Liposomal Clodronate

Song

Kim

Lee

et al. 2022

Tissue Eng Regen Med

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BACKGROUND: The use of mouse bone marrow mesenchymal stem cells (mBMSCs) represents a promising strategy for performing preclinical studies in the field of cell-based regenerative medicine; however, mBMSCs obtained via conventional isolation methods have two drawbacks, i.e., (i) they are heterogeneous due to frequent macrophage contamination, and (ii) they require long-term culturing for expansion. METHODS: In the present study, we report a novel strategy to generate highly pure mBMSCs using liposomal clodronate. This approach is based on the properties of the two cell populations, i.e., BMSCs (to adhere to the plasticware in culture dishes) and macrophages (to phagocytose liposomes). RESULTS: Liposomal clodronate added during the first passage of whole bone marrow culture was selectively engulfed by macrophages in the heterogeneous cell population, resulting in their effective elimination without affecting the MSCs. This method allowed the generation of numerous high-purity Sca-1+CD44+F4/80− mBMSCs (> 95%) with just one passaging. Comparative studies with mBMSCs obtained using conventional methods revealed that the mBMSCs obtained in the present study had remarkably improved experimental utilities, as demonstrated by in vitro multilineage differentiation and in vivo ectopic bone formation assays. CONCLUSION: Our newly developed method, which enables the isolation of mBMSCs using simple and convenient protocol, will aid preclinical studies based on the use of MSCs.

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