A replicating stem-like cell that contributes to bone morphogenetic protein 2-induced heterotopic bone formation.
A replicating stem-like cell that contributes to bone morphogenetic protein 2-induced heterotopic bone formation.
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DOI：
10.1002/sctm.20-0378
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发表时间：
2021-04
影响因子：
6
通讯作者：
Davis AR
中科院分区：
文献类型：
--
作者：
Mejia J;Salisbury E;Sonnet C;Gugala Z;Olmsted-Davis EA;Davis AR
骨形态发生蛋白2（BMP 2）诱导的异位骨形成（HBF）在BMP 2诱导后从零开始同步，这有利于谱系追踪。在BMP 2诱导的HBF期间，在GLAST‐ CreErt 2：td番茄红（TR）STOPflox小鼠中报告的研究显示，大约1周后，78.8 ± 11.6%的软骨细胞和成骨细胞为TR+。单细胞RNAseq后的聚类产生了9种细胞类型，分析显示其中一种是高度复制的干细胞样细胞（RSC）。假定时表明RSC转变为同时表达多种成骨细胞和软骨细胞转录物的间充质干细胞样细胞（软骨-骨祖细胞[COP]）。使用流式细胞术分离RSC和COP以获得独特的表面标志物。将分离的RSC（GLAST-TR + Hmmr+ Cd 200 −）和COP（GLAST-TR + Cd 200 + Hmmr−）注射到经历HBF的小鼠的肌肉中。在接受RSC的小鼠中，异位骨（HB）中约9%的细胞为GLAST-TR+，而接受COP的小鼠中不到0.5%的细胞为GLAST-TR+，这表明RSC的效力比COP高出许多倍。从移植HB中分离的供体来源TR+ RSC的分析显示，约50%为COP，45%为其他细胞，推测为成熟骨细胞，证实了RSC的早期性质。接下来，我们从这些小鼠（约300只）中分离出RSC，并将其注射到第二只动物中，在分析HBF时发现了类似的结果。与其他方法不同，单细胞RNAseq能够检测稀有细胞群，如RSC。RSCs可以注射到小鼠体内并分化的事实表明它们对组织再生的潜在效用。蓝色箭头显示骨形态发生蛋白2诱导后异位骨形成的轨迹。复制干细胞样细胞（RSC）是一种上皮细胞，其在形成表达成骨细胞和软骨细胞转录物的软骨-骨祖细胞（COP）时经历上皮-间充质转化。红色箭头显示TR+ RSC或TR+ COP的荧光激活细胞分选分离，并将3000 μ L每种TR+细胞注射到经历异位骨化的野生型小鼠中。
Bone morphogenetic protein 2 (BMP2)‐induced heterotopic bone formation (HBF) starts synchronously from zero upon BMP2 induction, which is advantageous for lineage tracking. The studies reported here in GLAST‐CreErt2:tdTomato red (TR)floxSTOPflox mice during BMP2‐induced HBF show 78.8 ± 11.6% of chondrocytes and 86.5 ± 1.9% of osteoblasts are TR+ after approximately 1 week. Clustering after single‐cell RNAseq resulted in nine cell types, and analysis revealed one as a highly replicating stem‐like cell (RSC). Pseudotiming suggested that the RSC transitions to a mesenchymal stem‐like cell that simultaneously expresses multiple osteoblast and chondrocyte transcripts (chondro‐osseous progenitor [COP]). RSCs and COPs were isolated using flow cytometry for unique surface markers. Isolated RSCs (GLAST‐TR+ Hmmr+ Cd200−) and COPs (GLAST‐TR+ Cd200+ Hmmr−) were injected into the muscle of mice undergoing HBF. Approximately 9% of the cells in heterotopic bone (HB) in mice receiving RSCs were GLAST‐TR+, compared with less than 0.5% of the cells in mice receiving COPs, suggesting that RSCs are many times more potent than COPs. Analysis of donor‐derived TR+ RSCs isolated from the engrafted HB showed approximately 50% were COPs and 45% were other cells, presumably mature bone cells, confirming the early nature of the RSCs. We next isolated RSCs from these mice (approximately 300) and injected them into a second animal, with similar findings upon analysis of HBF. Unlike other methodology, single cell RNAseq has the ability to detect rare cell populations such as RSCs. The fact that RSCs can be injected into mice and differentiate suggests their potential utility for tissue regeneration. Blue arrows show the trajectory of heterotopic bone formation after bone morphogenetic protein 2 induction. The replicating stem‐like cell (RSC) is an epithelial cell that undergoes an epithelial‐mesenchymal transition in forming the chondro‐osseous progenitor (COP) that expresses both osteoblast and chondrocyte transcripts. Red arrows show the fluorescence‐activated cell sorting isolation of either TR+ RSCs or TR+ COPs and the injection of 3000 of each TR+ cell into wild‐type mice undergoing heterotopic ossification.
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影响因子：
4.2
作者：
Fouletier-Dilling, Christine M.;Gannon, Francis H.;Davis, Alan R.
通讯作者：
Davis, Alan R.
DOI：
10.1038/bonekey.2013.171
发表时间：
2013-10-02
期刊：
BoneKEy reports
影响因子：
--
作者：
Guntur, Anyonya R;Rosen, Clifford J
通讯作者：
Rosen, Clifford J
DOI：
10.1073/pnas.0504750102
发表时间：
2005-10-11
影响因子：
11.1
作者：
Akiyama, H;Kim, JE;de Crombrugghe, B
通讯作者：
de Crombrugghe, B
影响因子：
5.1
作者：
Gugala, Z;Olmsted-Davis, EA;Davis, AR
通讯作者：
Davis, AR
影响因子：
6
作者：
Olmsted-Davis EA;Salisbury EA;Hoang D;Davis EL;Lazard Z;Sonnet C;Davis TA;Forsberg JA;Davis AR
通讯作者：
Davis AR