文献类型：期刊文献

英文题名：EX VIVO ISOLATION AND EXPANSION OF RABBIT MESENCHYMAL STEM CELLS IN DEFINED SERUM-FREE MEDIA

作者：Jin, Lifang[1,2];Hua, Yunfen[2];Cao, Huanhuan[2];Zhang, Ming[2]

机构：[1]Shaoxing Univ, Coll Life Sci, Shaoxing, Zhejiang, Peoples R China;[2]Hangzhou Precis Med Res Ctr, Hangzhou, Zhejiang, Peoples R China

年份：2018

卷号：34

期号：3

起止页码：811

外文期刊名：ACTA MEDICA MEDITERRANEA

收录：SCI-EXPANDED(收录号：WOS:000434980000028)、、Scopus(收录号：2-s2.0-85047330769)、WOS

基金：This work was supported by research grants from the Zhejiang Province Science and Technology Project of China [grant number 2013C33189].

语种：英文

外文关键词：Mesenchymal stem cells; Serum-free medium; Expansion; Differentiation; Rabbit

外文摘要：Introduction: Mesenchymal stem cells (MSCs) derived from rabbit bone marrow are routinely investigated in cardiovascular and orthopedic models for regenerative medicine. However, classical media used for generating MSCs are typically augmented with ill-defined supplements, such as fetal bovine serum, which increases safety concerns as well as inconsistency in MSC generation. We tested three commercial serum-free media for the generation and expansion of rabbit MSCs (rbMSCs). Materials and methods: RbMSCs were separated by adherent tissue culture and cultured in serum-free or serum-containing media. Cultured cells were characterized by phase contrast microscopy, cell-proliferation, flow cytometry, and qPCR analyses. Results: MesenCult (R) XF medium supported the generation and expansion of rbMSCs, whereas StemPro (R) hMSC and BD Mosaic hMSC media did not propagate freshly isolated rbMSCs. At passage 5, total cell yields for serum-containing and MesenCult (R) XF media were 4.0 +/- 0.16 x 108 and 1.9 +/- 0.28 x 109, and cumulative population doublings were 11.5 +/- 0.12 and 13.08 +/- 0.19, respectively. Colony formation efficiencies in MesenCult (R) XF were 21 to 26%, whereas those in serum-containing medium were 14 to 17%. Flow cytometry showed that rbMSCs in MesenCult (R) XF were positive for CD29, CD44 and CD73, and negative for CD34 and CD45. After corresponding differentiation, rbMSCs from MesenCult (R) XF could differentiate into adipogenic, osteogenic and chondrogenic lineages. qPCR analysis demonstrated that cells cultured in serum-free media had higher differentiation potentials than those cultured in serum-containing media. Discussion: rbMSCs were efficiently generated and expanded in MesenCult (R) XF, while maintaining the necessary characteristics attributed to MSCs for potential therapeutic use.