Placenta mesenchymal stem cell multi-source extraction system based on integrated processing platform

Abstract

The invention particularly relates to the technical field of stem cell preparation and tissue engineering, and discloses a placenta mesenchymal stem cell multi-source extraction system based on an integrated treatment platform. Comprising a multi-region synchronous pretreatment module, a zoning adherent culture module, a differentiated cell harvesting module, a cell characteristic collection module, a functional cell division module and a whole-process closed-loop control module, the multi-region synchronous pretreatment module adopts an integrated design, and the zoning adherent culture module realizes centralized culture through an independent marking region; the differentiation cell harvesting module is used for carrying out merging amplification; the cell characteristic acquisition module is used for acquiring cell characteristic data; the functional cell division module is used for realizing cell type and function judgment; the whole-process closed-loop control module is used for realizing whole-process quality tracing; through the integrated design, the cost is reduced, the cell quality stability is ensured, and the whole-process quality tracing is realized.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of stem cell preparation and tissue engineering, and more particularly to a placental mesenchymal stem cell multi-source extraction system based on an integrated processing platform. BACKGROUND

[0002] As an important supporting organ during the fetal period, the placenta contains a variety of mesenchymal cells with high differentiation potential and immunoregulatory function. Mesenchymal stem cells have sub-fully differentiated potential and can be induced to differentiate into various tissue cells in a specific environment. Placental mesenchymal stem cells have strong regenerative capacity, early development, weak immunogenicity, and large tissue compatibility. Placental mesenchymal stem cells have become an important source of mesenchymal stem cells.

[0003] Traditional placental mesenchymal stem cell extraction mostly uses the tissue block adherence method. After the placental tissue is sterilized, impurities are removed, serum-containing medium is added, and the culture box is placed in the culture box for static culture. The cells migrate out of the tissue block and proliferate.

[0004] However, in actual use, there are still some shortcomings. First, single source leads to functional limitations. Traditional placental mesenchymal stem cell extraction only extracts mesenchymal stem cells from a single tissue area, which can only obtain a relatively limited functional stem cell population, making it difficult to meet the requirements of personalized treatment and limiting the application of mesenchymal stem cells in clinical medicine. Second, the cell yield is low and the cost is high. Traditional placental mesenchymal stem cell extraction does not fully utilize placental tissue resources, resulting in serious resource waste. The extraction of each region is independent, and there is a lack of effective system utilization mechanism. The relatively dispersed extraction method is difficult to ensure the cell yield, and the quality of the extracted mesenchymal stem cells is not uniform. Third, there is a lack of a closed-loop management system. Traditional placental mesenchymal stem cell extraction is highly dependent on the manual experience of the operator. Differences in operator experience may lead to deviations. It is difficult to form a unified standard and specification during the extraction process, making it difficult to control the quality of the cells and not conducive to quality tracing of the mesenchymal stem cell extraction process. SUMMARY

[0005] Therefore, the embodiment of the present application provides a placental mesenchymal stem cell multi-source extraction system based on an integrated processing platform, which adopts an integrated design through a multi-region synchronous pretreatment module, realizes centralized culture through an independent marking area of a partitioned adherent culture module, performs combined expansion through a differential cell harvesting module, realizes cell type and function determination through a cell characteristic acquisition module and a functional cell division module, and realizes whole-process quality tracing through a whole-process closed-loop control module, thereby effectively solving the problems of functional limitation caused by single source, low cell yield and high cost, and lack of closed-loop management system in the background art.

[0006] To achieve the above object, the present application provides the following technical scheme: a placental mesenchymal stem cell multi-source extraction system based on an integrated processing platform, comprising a multi-region synchronous pretreatment module, a partitioned adherent culture module, a differential cell harvesting module, a cell characteristic acquisition module, a functional cell division module, and a whole-process closed-loop control module: The multi-region synchronous pretreatment module is used for separating the tissue regions of fresh placenta, uniformly pretreating the placental tissue regions, and obtaining cell tissue samples corresponding to each placental tissue region. The partitioned adherent culture module is used for adopting a multi-region marked adherent culture plate, adding a human serum-free culture medium, and culturing the cell tissue samples of each placental tissue region in a partitioned manner. The differential cell harvesting module is used for observing the cell migration and adhesion state in a partitioned manner, sucking the adherent cells, transferring them to a culture bottle, and performing combined expansion culture to construct a mixed cell system. The cell characteristic acquisition module is used for obtaining cell characteristic data of the target detection cells in the culture bottle, including cell phenotype data and cell activity function data. The functional cell division module comprises a cell verification unit and a cell division unit, and is used for dividing the target detection cell type and function based on the cell characteristic data. The whole-process closed-loop control module is used for monitoring the operation parameters and detection data of each module in real time, performing deviation determination based on a preset standard, and triggering a correction mechanism if a deviation occurs.

[0007] The technical effects and advantages of the present application are as follows: 1. The present application jointly extracts the amnion, chorion, decidua and umbilical cord attachment region of the placenta through a multi-region synchronous processing module, determines the type and function of the cells in different regions through a functional cell division module, obtains a mixed system of mesenchymal stem cells with various functional differences, can meet the individualized needs of treatment, and effectively expands the application range of mesenchymal stem cells. 2、The placental mesenchymal stem cell multi-source extraction system based on the integrated processing platform, including a multi-region synchronous pretreatment module, a partitioned adherent culture module, a differential cell harvesting module, a cell characteristic acquisition module, a functional cell division module and a whole-process closed-loop control module. 3、The placental mesenchymal stem cell multi-source extraction system based on the integrated processing platform, including a multi-region synchronous pretreatment module, a partitioned adherent culture module, a differential cell harvesting module, a cell characteristic acquisition module, a functional cell division module and a whole-process closed-loop control module. BRIEF DESCRIPTION OF DRAWINGS

[0008] Figure 1 It is the overall structure schematic diagram of the present application.

[0009] Figure 2 It is the cell tissue sample acquisition step schematic diagram of the present application.

[0010] Figure 3 It is the cell tissue sample partitioned culture step schematic diagram of the present application. DETAILED DESCRIPTION

[0011] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0012] As shown in the placental mesenchymal stem cell multi-source extraction system based on the integrated processing platform, including a multi-region synchronous pretreatment module, a partitioned adherent culture module, a differential cell harvesting module, a cell characteristic acquisition module, a functional cell division module and a whole-process closed-loop control module. Figure 1 The placental mesenchymal stem cell multi-source extraction system based on the integrated processing platform, including a multi-region synchronous pretreatment module, a partitioned adherent culture module, a differential cell harvesting module, a cell characteristic acquisition module, a functional cell division module and a whole-process closed-loop control module.

[0013] The specific embodiments of the present application include the following contents: The multi-region synchronous pretreatment module: separate the tissue regions of fresh placenta, uniformly pretreat the placental tissue regions, and obtain the cell tissue samples corresponding to each placental tissue region; Further, the placental tissue region includes amniotic membrane layer, chorionic membrane, iris tissue and umbilical cord attachment part.

[0014] In this embodiment, it needs to be specifically pointed out that the separation of the four placental tissue regions needs to be carried out in a sterile clean environment, and the fresh placenta refers to the placenta collected within two hours. During the dissection process, it is necessary to ensure that each tissue region is not mixed, so as to provide pure samples for subsequent targeted treatment.

[0015] The amniotic membrane layer is one of the important sources of mesenchymal stem cells, has advantages in immune regulation function, and can be involved in the research and treatment of immune-related diseases; the chorionic membrane is a tissue source for extracting mesenchymal stem cells with osteogenic potential, and has application value in the medical field of bone injury repair; the mesenchymal stem cells in the iris tissue can be used in the research of nerve repair; the mesenchymal stem cells in the umbilical cord attachment part can form a functional complement with other area cells, and can improve the treatment universality of the stem cell population through multi-area joint extraction.

[0016] Further, as shown in Figure 2 , the steps of obtaining the cell tissue sample are as follows: S1.1: using sterile instruments to cut each placental tissue region into 1mm 3 tissue blocks; In this embodiment, it needs to be specifically pointed out that cutting each placental tissue region into 1mm 3 tissue blocks can not only ensure the integrity of the tissue and maintain cell activity, but also increase the contact area between the tissue and the subsequent treatment liquid, which is beneficial to subsequent cleaning and potential enzymatic operation.

[0017] S1.2: Place the tissue blocks corresponding to each placental tissue region in a sterile solution, add PBS buffer solution, and rinse 3 times; In this embodiment, it needs to be specifically pointed out that the tissue blocks are rinsed 3 times to wash away the blood stains, mucus and impurities on the surface of the tissue through physical rinsing, so as to reduce the interference on the subsequent cell extraction.

[0018] S1.3: Transfer the cleaned tissue blocks to a centrifuge tube containing sterile human low-osmotic balance solution and gently shake; In this embodiment, it needs to be specifically pointed out that through gentle shaking, the low-osmotic balance solution can fully penetrate the tissue, destroy the residual red blood cell membrane by using the osmotic pressure difference, realize efficient removal of blood components, and avoid damage to the activity of mesenchymal stem cells.

[0019] S1.4: Selectively use 0.1% collagenase I to gently lyse the tissue blocks.

[0020] In this embodiment, it needs to be specifically pointed out that the mild lysis specifically refers to mixing the enzyme solution with the tissue block, reacting for 10 minutes under the condition of 37°C, destroying the interstitial structure of the tissue through the specific hydrolysis of the enzyme, promoting the mesenchymal stem cells to be separated from the tissue, and then immediately adding the serum-containing termination solution to terminate the enzyme lysis reaction, and repeatedly washing with PBS buffer to completely remove the residual enzyme solution, avoiding the excessive action of the enzyme to cause the decrease of the cell activity.

[0021] The partitioned adherent culture module: a multi-region marked adherent culture plate is used, human serum-free culture medium is added, and the cell tissue samples of each placental tissue region are cultured in partitions; Further, as shown in Figure 3 The steps of the partitioned culture of the cell tissue samples are as follows: S2.1: The multi-region marked adherent culture plate is divided into four independent marked regions, and is marked as A, B, C and D regions, which respectively correspond to each placental tissue region; S2.2: The cell tissue samples are inoculated into the corresponding independent marked regions, and the human serum-free culture medium is added into each independent marked region according to the unified standard; In this embodiment, it needs to be specifically pointed out that the cell tissue samples in each independent marked region are uniformly distributed, which ensures sufficient contact with the surface of the culture plate and provides a basis for cell migration and adhesion, and the amount of the culture medium is set according to the area of each independent marked region, which is used to ensure that the tissue blocks are covered and the liquid level depth is consistent, avoiding the uneven cell growth caused by the difference in nutrient supply.

[0022] S2.3: The multi-region marked adherent culture plate is placed in a CO2 incubator to maintain a stable temperature environment and CO2 concentration, and the humidity in the incubator is kept ≥95%; In this embodiment, it needs to be specifically pointed out that maintaining a stable temperature environment means setting the temperature in the incubator to 37°C ± 0.5°C, and the CO2 concentration in the incubator is maintained at 5% ± 0.2%, which is used to provide stable physiological growth conditions for cells and promote the migration and adhesion of mesenchymal stem cells in the tissue blocks.

[0023] S2.4: After the multi-region marked adherent culture plate is left to adhere for 48 hours, the liquid in each independent marked region is replaced daily.

[0024] In this embodiment, it needs to be specifically pointed out that during the adhesion, the culture plate should not be moved to ensure that the tissue blocks are tightly combined with the surface of the culture plate to create attachment conditions for cell migration; the liquid replacement operation means removing the old culture medium and adding fresh human serum-free culture medium according to the original standard, which is used to remove metabolic waste and supplement nutrients to maintain the stability of the cell growth environment.

[0025] Differential cell harvesting module: observe the cell migration and adhesion state in different regions, suck the adherent cells, transfer to culture bottles, and perform combined expansion culture to construct a mixed cell system. Further, the construction steps of the mixed cell system are as follows: S3.1: On the 5th day after the cell tissue sample is inoculated into the multi-region marked adherent culture plate, observe the cell state in the A, B, C, and D independent marked regions by microscope every day, and record the number, morphology, and adhesion density of the migrated cells around the tissue block; In this embodiment, it needs to be specifically pointed out that when recording the state of the migrated cells around the tissue block, it is necessary to monitor whether the cells appear abnormal growth at the same time, including but not limited to contamination and apoptosis. In the observation process, the regions should be strictly distinguished to ensure the accuracy of the judgment of the growth characteristics of different source cells, and to provide a basis for the selection of the subsequent harvesting time.

[0026] S3.2: After the cell tissue sample is inoculated into the multi-region marked adherent culture plate for 7 days, when the adherent cells in each independent marked region reach the logarithmic growth phase and the morphology is uniform, perform partition harvesting; In this embodiment, it needs to be specifically pointed out that the partition harvesting specifically refers to gently sucking the adherent cells from the A, B, C, and D four independent marked regions by using a sterile pipette, avoiding disturbing the un-migrated tissue block and un-adherent cells, ensuring that the cells in each region are collected separately and do not cross-contaminate, and completely retaining the population characteristics of different source cells.

[0027] S3.3: Transfer the cell suspensions harvested from the four independent marked regions to a unified sterile culture bottle, add fresh human serum-free culture medium, and place it in a culture box for joint expansion to obtain a mixed cell population and construct a multi-functional cell system.

[0028] In this embodiment, it needs to be specifically pointed out that the acquisition of the mixed cell population needs to observe the cell growth state every day during the expansion process. When the cell fusion degree reaches 80%-90%, passaging is performed to obtain sufficient mixed cell population. The multi-functional cell system includes amniotic mesenchymal stem cells, chorionic mesenchymal stem cells, decidua mesenchymal stem cells, and umbilical cord attachment mesenchymal stem cells.

[0029] Cell characteristic acquisition module: acquire cell characteristic data of the target detection cells in the culture bottle, including cell phenotype data and cell activity function data; Further, the cell phenotype data includes positive marker expression rate and negative marker expression rate, and the cell activity function data includes total cell number, viable cell ratio, osteogenic differentiation positive cell rate, adipogenic differentiation positive cell rate, and chondrogenic differentiation positive cell rate.

[0030] In this embodiment, it needs to be specifically pointed out that the collection of cell characteristic data needs to be carried out after the completion of the expansion culture, corresponding to the 8-14 days of the multi-source extraction process of placental mesenchymal cells.

[0031] The positive marker expression rate includes the expression rates of CD73⁺, CD90⁺ and CD105⁺, and the negative marker expression rate includes the expression rates of CD45⁻ and CD34⁻. The positive marker is a specific expression marker of mesenchymal stem cells, and the negative marker is used to exclude the pollution of hematopoietic cells and other mixed cells. Flow cytometry is used to detect cell surface markers.

[0032] Cell counting method is used to count the total number of cells and the proportion of living cells, which is used to evaluate the cell proliferation ability. Fat, osteogenic and chondrogenic differentiation induction experiments are carried out on the cells respectively. Specific staining is used, including oil red O staining to identify fat differentiation, alizarin red staining to identify osteogenic differentiation and alcin blue staining to identify chondrogenic differentiation. The positive cell rate of each differentiation direction is calculated to quantify the multi-directional differentiation potential of the cells.

[0033] The functional cell division module includes a cell verification unit and a cell division unit, which divides the target detection cell type and function based on the cell characteristic data; Further, the cell verification unit is used to verify whether the target detection cell meets the general properties of mesenchymal stem cells. The positive marker score S + and the negative marker score S - are calculated according to the cell phenotype data, and the positive marker score and the negative marker score are calculated by the formula to obtain the comprehensive verification index SI of mesenchymal stem cells, wherein μ1 and μ2 are weight coefficients, the sum is 1, and the judgment standard is: when , the cell meets the general properties of mesenchymal stem cells.

[0034] In this embodiment, it needs to be specifically pointed out that the weight coefficients of the positive marker score and the negative marker score can be 0.6 and 0.4. The weight of the positive marker is greater than that of the negative marker, and the positive marker is the core identification feature of mesenchymal stem cells.

[0035] The calculation steps of the positive marker score are as follows: The expression rates of CD73⁺, CD90⁺ and CD105⁺ are introduced into the formula: , to calculate the positive marker score S + . Through natural logarithmic conversion, the positive contribution of high expression rate is amplified, and through the logarithmic total value of 100% expression rate, the value range of the positive marker score is normalized to 0-1. The calculation steps of the negative marker score are as follows: The expression rate of CD45⁻ and CD34⁻ is introduced into the formula: , The negative marker score S - is calculated. The positive contribution of low expression rate is amplified by an exponential function, and is normalized by an exponential total value of 5%, so that the value range of the negative marker score is 0-1.

[0036] When , the cell phenotype meets the general properties of mesenchymal stem cells, and enters the subsequent functional division link; when , it means that the cell phenotype does not meet the general properties, and is determined as a non-target mesenchymal stem cell, which needs to trace back the operation deviation of the previous link.

[0037] Further, the cell division unit is used to calculate the cell differentiation characteristic matching degree, judge the cell type and cell function, and the calculation steps of the cell differentiation characteristic matching degree are as follows: S4.1: According to the total number of cells N and the proportion of living cells R, the cell activity index D V is calculated by the formula According to the osteogenic differentiation positive cell rate O, the adipogenic differentiation positive cell rate F and the chondrogenic differentiation positive cell rate G, the osteogenic differentiation index D O , the adipogenic differentiation index D F and the chondrogenic differentiation index D G are calculated by the formula , and the value of X is O, F and G; In this embodiment, it needs to be specifically pointed out that in the calculation process of the cell activity index, the contribution of the high activity cell population is amplified by natural logarithmic conversion, and 1 is added to avoid zero value interference. In the calculation process of the differentiation index, the weight of the high differentiation positive rate is strengthened by the exponential function, which is conducive to making the dominant differentiation characteristics more significant.

[0038] S4.2: Obtain the regional cell function characteristic reference value, calculate the matching degree M i of the cell activity index, the osteogenic differentiation index, the adipogenic differentiation index and the chondrogenic differentiation index of the target detection cell with the target type reference value, and the value of i is V, O, F and G; In this embodiment, it needs to be specifically pointed out that the preset reference value can be: Cell type Cell activity index Osteogenic differentiation index Adipogenic differentiation index Chondrogenic differentiation index Amnion 4.5-5.0 1.0-1.6 1.7-2.2 1.4-1.8 Chorion 4.3-4.8 2.0-2.7 0.8-1.3 1.0-1.5 Decidua 4.0-4.5 1.3-1.8 1.0-1.5 1.8-2.3 Cord insertion 4.3-4.8 1.5-2.0 1.3-1.8 1.3-1.8 Table 1-1

[0039] The calculation steps of the matching degree of the target detection cell and the target type reference value are as follows: The cell activity index, the osteogenic differentiation index, the adipogenic differentiation index and the chondrogenic differentiation index are calculated by the formula: ,

[0040] The matching degree M of the target detection cell and the target type reference value is calculated i , wherein i is V, O, F, and G, the reference median value, the reference maximum value, and the reference minimum value are obtained according to the preset reference values in Table 1-1, the matching degree has a value range of 0-1, the closer to the reference median value, the higher the matching degree score, when the value of the matching degree is greater than or equal to 0, M i , otherwise, 0.

[0041] S4.3: Substitute the cell activity index matching degree, the osteogenic differentiation index matching degree, the adipogenic differentiation index matching degree, and the chondrogenic differentiation index matching degree into the formula: , The cell differentiation characteristic matching degree DI is calculated, wherein ω1, ω2, ω3, and ω4 respectively represent the weight coefficients of the cell activity index matching degree, the osteogenic differentiation index matching degree, the adipogenic differentiation index matching degree, and the chondrogenic differentiation index matching degree, and the sum is 1.

[0042] In this embodiment, it needs to be specifically explained that the weight coefficients of the cell activity index matching degree, the osteogenic differentiation index matching degree, the adipogenic differentiation index matching degree, and the chondrogenic differentiation index matching degree can be 0.1, 0.3, 0.3, and 0.3, and the weights of the osteogenic differentiation index matching degree, the adipogenic differentiation index matching degree, and the chondrogenic differentiation index matching degree should be greater than the weight of the cell activity index matching degree, so as to highlight the influence of differentiation function on the matching degree.

[0043] The cell differentiation characteristic matching degree includes the amniotic layer, chorionic membrane, iris tissue, and umbilical cord attachment part tissue area cell differentiation characteristic matching degree, which are respectively represented as DI1, DI2, DI3, and DI4, representing the matching degree of the target detection cell and the amniotic layer, chorionic membrane, iris tissue, and umbilical cord attachment part tissue area cell.

[0044] Further, the judgment of the cell type needs to set the matching degree reference value DI 标 , compare the cell differentiation characteristic matching degree of the target detection cell with the comprehensive matching degree reference value, and judge whether it meets the cell type standard, when multiple cell type standards are met, the cell type with the highest matching degree is taken, and if the cell differentiation characteristic matching degrees of the detection cell and the four kinds of placental tissue area cells are the same, the cell source area is combined for auxiliary judgment. The judgment of the cell function needs to calculate the differentiation potential contribution value of the osteogenic differentiation, adipogenic differentiation, and chondrogenic differentiation in the detection cell, including ω2×O, ω3×F, and ω4×G, and the differentiation direction with the maximum differentiation potential contribution value is the core differentiation function of the cell.

[0045] In this embodiment, it needs to be specifically pointed out that the matching characteristic degrees DI1, DI2, DI3 and DI4 of the target detection cells and the amniotic membrane layer, chorion, iris tissue and umbilical cord attachment region cells are compared with the comprehensive matching degree reference values of each type, and the cell type is judged, wherein the comprehensive matching degree reference value is obtained by taking the reference median value in the preset reference value table 1-1 through the above-mentioned cell differentiation characteristic matching degree calculation formula, and is expressed as DI 1标 , DI 2标 , DI 3标 , and DI 4标 .

[0046] The whole-process closed-loop control module: real-time monitoring of each module operation parameter and detection data, deviation judgment based on preset standard, if deviation triggers correction mechanism.

[0047] Further, the whole-process closed-loop control module connects the multi-region synchronous pretreatment module, the partitioned adherent culture module, the differential cell harvesting module, the cell characteristic acquisition module and the functional cell division module through the Internet of Things interface, and real-time acquisition of operation parameters and detection data of each module; In this embodiment, it needs to be specifically pointed out that the data of the multi-region synchronous pretreatment module includes but is not limited to placenta tissue cutting size, PBS cleaning times and hypotonic equilibrium liquid treatment time; the data of the partitioned adherent culture module includes but is not limited to the region correspondence of multi-region marked adherent culture plate, the addition amount of human serum-free medium, the temperature of the incubator and the CO2 concentration of the incubator; the data of the differential cell harvesting module includes but is not limited to cell observation starting time, harvesting time, cell transfer amount of each region and combined expansion record; the data of the cell characteristic acquisition module includes but is not limited to expression rate, cell count result and differentiation staining positive rate; the data of the functional cell division module includes but is not limited to mesenchymal stem cell comprehensive verification index and cell differentiation characteristic matching degree.

[0048] The deviation judgment needs to determine the operation parameter deviation and the detection data deviation according to the standardized threshold system, construct the deviation correction mechanism and trace the process.

[0049] In this embodiment, it needs to be specifically pointed out that the operation parameter deviation, for example, when the harvesting time is earlier than the 6th day or later than the 8th day, is judged as the harvesting opportunity deviation; the detection data deviation, for example, when the total yield of single fetus cells is lower than 80% of the historical average, is judged as abnormal yield; the correction mechanism is to automatically send calibration instructions to the parameters that can be adjusted in time and push prompt information to the manual operation link to guide the operation personnel to intervene; the process tracing refers to locating the deviation source module in reverse through the data chain, and automatically freezing the output process of the cell population judged as unqualified, only allowing the cells passing through the deviation correction and review to enter the subsequent application, and recording the correction measures and results.

[0050] Secondly: the embodiment of the present application discloses only the structure related to the embodiment of the present application, other structures can refer to the general design, and in the case of no conflict, the same embodiment and different embodiments of the present application can be combined with each other; Finally: the above only describes the preferred embodiments of the present application and is not used to limit the present application, any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.

Claims

1. A multi-source extraction system for placental mesenchymal stem cells based on an integrated processing platform, characterized in that: It includes a multi-region synchronous pretreatment module, a zoned adherent culture module, a differentiated cell harvesting module, a cell feature acquisition module, a functional cell classification module, and a closed-loop control module for the entire process: The multi-region synchronous preprocessing module is used to separate the tissue regions of the fresh placenta, perform uniform preprocessing on the placental tissue regions, and obtain cell tissue samples corresponding to each placental tissue region. The partitioned adherent culture module is used to culture cell tissue samples from each placental tissue region in a partitioned manner using a multi-region labeled adherent culture plate and human serum-free culture medium. The differential cell harvesting module is used to observe cell migration and adhesion in different regions, aspirate adhered cells, transfer them to culture flasks, and perform combined amplification culture to construct a mixed cell system. The cell feature acquisition module is used to acquire cell feature data of the target cells in the culture flask, including cell phenotype data and cell activity and function data. The functional cell segmentation module is used to include a cell verification unit and a cell segmentation unit, and to segment the target detection cell type and function based on cell feature data. The full-process closed-loop control module is used to monitor the operating parameters and detection data of each module in real time, determine deviations based on preset standards, and trigger a correction mechanism if a deviation occurs.

2. The placental mesenchymal stem cell multi-source extraction system based on an integrated processing platform according to claim 1, characterized in that: The placental tissue region includes the amnion layer, chorion, iris tissue, and umbilical cord attachment.

3. The placental mesenchymal stem cell multi-source extraction system based on an integrated processing platform according to claim 1, characterized in that: The steps for obtaining the cell tissue sample are as follows: S1.1: Using sterile instruments, each placental tissue region was cut into 1mm pieces. 3 The organizational blocks; S1.2: Place the tissue blocks corresponding to each placental tissue region in a sterile solution, add PBS buffer, and rinse 3 times; S1.3: Transfer the cleaned tissue block to a centrifuge tube containing sterile human hypotonic balanced solution and gently shake. S1.4: Selectively use 0.1% collagenase I to gently lyse tissue blocks.

4. The placental mesenchymal stem cell multi-source extraction system based on an integrated processing platform according to claim 1, characterized in that: The steps for culturing cell tissue samples in specific regions are as follows: S2.1: Divide the multi-region labeled adherent culture plate into 4 independent labeled regions, labeled as regions A, B, C and D, corresponding to each placental tissue region; S2.2: Seed the cell tissue samples into the corresponding independently labeled areas, and add human serum-free culture medium to each independently labeled area according to a unified standard; S2.3: Place the multi-region labeled adherent culture plate in a CO2 incubator, maintain a stable temperature environment and CO2 concentration, and keep the humidity inside the incubator ≥95%; S2.4: After the multi-region labeled culture plate has been statically attached to the wall for 48 hours, the medium should be changed daily for each individual labeled region.

5. The placental mesenchymal stem cell multi-source extraction system based on an integrated processing platform according to claim 1, characterized in that: The steps for constructing the hybrid cell system are as follows: S3.1: On the 5th day after cell tissue samples were seeded into multi-region labeled adherent culture plates, the cell status in the independently labeled regions A, B, C and D was observed daily by microscope, and the number, morphology and adhesion density of cells migrating out from around the tissue block were recorded. S3.2: On the 7th day after cell tissue samples were seeded into multi-region labeled adherent culture plates, when the adherent cells in each independent labeled region reached the logarithmic growth phase and were morphologically uniform, they were harvested in sections. S3.3: The cell suspensions harvested from the four independently labeled regions were transferred to a uniform sterile culture flask, fresh human serum-free culture medium was added, and the flasks were placed in an incubator for joint amplification to obtain a mixed cell population and construct a multifunctional cell system.

6. The placental mesenchymal stem cell multi-source extraction system based on an integrated processing platform according to claim 1, characterized in that: The cell phenotypic data include the expression rates of positive and negative markers, and the cell viability data include the total number of cells, the proportion of live cells, the rate of osteogenic differentiation positive cells, the rate of adipogenic differentiation positive cells, and the rate of chondrogenic differentiation positive cells.

7. The placental mesenchymal stem cell multi-source extraction system based on an integrated processing platform according to claim 1, characterized in that: The cell validation unit is used to verify whether the target cells conform to the general properties of mesenchymal stem cells, and calculates the positive marker score S based on the cell phenotype data. + and negative biomarker score S - And based on the positive and negative biomarker scores, through a formula The comprehensive validation index (SI) for mesenchymal stem cells was calculated, where μ1 and μ2 are weighted coefficients, and their sum is 1. The judgment criterion is: when... At that time, the cells met the general properties of mesenchymal stem cells.

8. The placental mesenchymal stem cell multi-source extraction system based on an integrated processing platform according to claim 1, characterized in that: The cell segmentation unit is used to calculate the cell differentiation feature matching degree to determine cell type and cell function. The calculation steps for the cell differentiation feature matching degree are as follows: S4.1: Based on the total number of cells N and the proportion of viable cells R, use the formula... The cell viability index D was calculated. V Based on the positive rates of osteogenic differentiation (O), adipogenic differentiation (F), and chondrogenic differentiation (G), the following formulas were used to... The osteogenic differentiation index D was calculated. O Adipogenic differentiation index D F and chondrogenic differentiation index D G X can take the values ​​O, F, and G; S4.2: Obtain the baseline values ​​of regional cell functional characteristics, and calculate the matching degree M between the target detection cells' cell viability index, osteogenic differentiation index, adipogenic differentiation index, and chondrogenic differentiation index and the baseline values ​​of the target type. i The values ​​of i are V, O, F, and G; S4.3: Substitute the matching degree of cell viability index, osteogenic differentiation index, adipogenic differentiation index, and chondrogenic differentiation index into the formula: ， The cell differentiation characteristic matching degree DI was calculated, where ω1, ω2, ω3 and ω4 represent the weight coefficients of cell activity index matching degree, osteogenic differentiation index matching degree, adipogenic differentiation index matching degree and chondrogenic differentiation index matching degree, respectively, and the sum is 1.

9. The multi-source extraction system for placental mesenchymal stem cells based on an integrated processing platform according to claim 1, characterized in that: The cell type determination requires setting a matching baseline value (DI). 标 The matching degree of the cell differentiation characteristics of the target cell is compared with the comprehensive matching degree benchmark value to determine whether it meets the standard of the cell type. When multiple cell type standards are met, the cell type with the highest matching degree is selected. If the matching degree of the cell differentiation characteristics of the target cell is the same as that of the cells in the four placental tissue regions, the cell source region is combined to assist in the determination. Determining cell function requires calculating the differentiation potential contribution values ​​of osteogenic, adipogenic, and chondrogenic differentiation in the cells, including ω2×O, ω3×F, and ω4×G. The differentiation direction with the largest differentiation potential contribution value is the core differentiation function of the cell.

10. The multi-source extraction system for placental mesenchymal stem cells based on an integrated processing platform according to claim 1, characterized in that: The full-process closed-loop control module is connected to the multi-region synchronous pretreatment module, the partitioned adherent culture module, the differentiated cell harvesting module, the cell feature acquisition module, and the functional cell classification module through the Internet of Things interface, and collects the operating parameters and detection data of each module in real time. Deviation determination requires identifying deviations in operating parameters and detection data based on a standardized threshold system, constructing a deviation correction mechanism, and performing process traceability.

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